Language：English   Publisher：シュプリンガー・ジャパン(株)  

食道平滑筋収縮の変化および関連する炎症性反応について検討することによりアカラシアの病態を明らかにし、アカラシアの進展における食道細菌叢の意義を評価した。経口的内視鏡的筋層切開術を施行したII型アカラシア症例から得た食道粘膜および下部食道括約筋(LES)標本を解析した。患者由来の食道調整培養液をマウス食道に投与し、食道内環境がアカラシアと関連するか否かについて検討した。安静時および刺激時における対照群のLESでは、20-kDaミオシン軽鎖(LC20)の約30%がリン酸化されていた。アカラシア症例では状況に関わらずLC20のリン酸化は10%未満であった。アカラシアにおける低リン酸化は、ミオシンリン酸化阻害蛋白質CPI-17の発現低下と関連していた。IL-17A、IL-17F、IL-22、IL-23Aを含む関連サイトカインは、アカラシアでは発現が増加していた。食道細菌叢のα-Diversity指数、およびActinomycesおよびDialisterを含む数種類の細菌の割合がアカラシアで増加していた。ActinomycesレベルはIL-23Aレベルと正の相関が認められ、DialisterレベルはIL-17A、IL-17F、IL-22レベルと正の相関が認められた。マウスにおいて、食道IL-17F値は調整培地の経口投与後に増加した。

Language：English  

Purpose: Cholelithiasis occurs often after gastrectomy. However, no consensus has been established regarding the difference in the incidence of postgastrectomy cholelithiasis with different reconstruction methods. In this study, we examined the frequency of cholelithiasis after two major reconstruction methods, namely Billroth-I (B-I) and Roux-en-Y (R-Y) following laparoscopic distal gastrectomy (LDG) for gastric cancer.
Methods: Among 696 gastric cancer patients who underwent LDG between April 2000 and March 2017, after applying the exclusion criteria, 284 patients who underwent B-I and 310 who underwent R-Y were examined retrospectively. The estimated incidence of cholelithiasis was compared between the methods, and factors associated with the development of cholelithiasis in the gallbladder and/or common bile duct were investigated.
Results: During the median follow-up of 61.2 months, 52 patients (8.8%) developed cholelithiasis postgastrectomy; 12 patients (4.2%) after B-I and 40 (12.9%) after R-Y (p = 0.0002). Among them, choledocholithiasis was more frequent in patients who underwent R-Y (n = 11, 27.5%) vs. B-I (n = 1, 8.3%) (p = 0.0056). Univariate and multivariate analyses revealed that male sex, body mass index > 22.5 kg/m2, and R-Y reconstruction were significant predictors of the development of postLDG cholelithiasis.
Conclusion: Regarding cholelithiasis development, B-I reconstruction should be preferred whenever possible during distal gastrectomy.
Keywords: Billroth-I; Cholelithiasis; Distal gastrectomy; Gastric cancer; Laparoscopy; Roux-en-Y.

DOI： 10.1007/s00423-024-03267-2

Language：English  

Preoperative treatment is commonly carried out for borderline resectable pancreatic ductal adenocarcinoma (PDAC). However, the relationship between the combination of immune cells in the tumor microenvironment and their intratumoral heterogeneity along with their association with histological findings remains unclear, especially in patients receiving preoperative chemotherapy. We aimed to explore the therapeutic strategies for patients with PDAC with poor prognosis after receiving chemotherapy based on histological and immunological microenvironmental classifications. We investigated the correlation between the prognosis and histological immune microenvironmental factors of patients who initially underwent surgery (n = 100) and were receiving gemcitabine plus nab-paclitaxel (GEM + nabPTX) as preoperative chemotherapy (n = 103). Immune profiles were generated based on immune cell infiltration into the tumor, and their correlation with patient outcomes and histological features was analyzed. Tumor-infiltrating neutrophils (TINs) were identified as independent poor prognostic factors using multivariate analysis in both surgery-first and preoperative chemotherapy groups. The patients were further classified into four groups based on immune cell infiltration into the tumor. Patients with high CD15 infiltration into the tumor and immature stroma at the cancer margins showed the worst prognosis in the preoperative chemotherapy group. The analysis of mRNA expression and immunohistochemical features revealed that CXCR2, the receptor for CXCL8, was correlated with disease-free and overall survival. We inferred that patients with immature stroma at the margins and high infiltration of CD15+ neutrophils within the tumor showed the worst prognosis and they could particularly benefit from treatment with inhibitors targeting CXCR2 or CXCL8.
Keywords: neoadjuvant chemotherapy; neutrophils; pancreatic cancer; receptors, chemokine; tumor microenvironment.

DOI： 10.1111/cas.16060

Language：English  

Background: Pancreatic ductal adenocarcinoma (PDAC) has a high mortality rate owing to its late diagnosis and aggression. In addition, there are relatively few minimally invasive screening methods for the early detection of PDAC, making the identification of biomarkers for this disease a critical priority. Recent studies have reported that microRNAs in extracellular vesicles (EV-miRs) from bodily fluids can be useful for the diagnosis of PDACs. Given this, we designed this study to evaluate the utility of cancer EVs extracted from duodenal fluid (DF) and their resident EV-miRs as potential biomarkers for the detection of PDAC.

Methods: EV-miRs were evaluated and identified in the supernatants of various pancreatic cancer cell lines (Panc-1, SUIT2, and MIAPaca2), human pancreatic duct epithelial cells, and the DF from patients with PDAC and healthy controls. EVs were extracted using ultracentrifugation and the relative expression of EV-miR-20a was quantified.

Results: We collected a total of 34 DF samples (27 PDAC patients and seven controls) for evaluation and our data suggest that the relative expression levels of EV-miR-20a were significantly higher in patients with PDAC than in controls (p = 0.0025). In addition, EV-miR-20a expression could discriminate PDAC from control patients regardless of the location of the tumor with an area under the curve values of 0.88 and 0.88, respectively.

Conclusions: We confirmed the presence of EVs in the DF and suggest that the expression of EV-miR-20a in these samples may act as a potential diagnostic biomarker for PDAC.

Keywords: duodenal fluid; early detection; extracellular vesicle; microRNA; pancreatic ductal adenocarcinoma.

DOI： 10.1002/deo2.333

Language：English  

Background
Gastric cancer (GC) is characterized by an immunosuppressive and treatment-resistant tumor immune microenvironment (TIME). Here, we investigated the roles of different immunosuppressive cell types in the development of the GC TIME.
Methods
Single-cell RNA sequencing (scRNA-seq) and multiplex immunostaining of samples from untreated or immune checkpoint inhibitor (ICI)-resistant GC patients were used to examine the correlation between certain immunosuppressive cells and the prognosis of GC patients.
Results
The results of the scRNA-seq analysis revealed that tumor-infiltrating monocytic myeloid-derived suppressor cells (TI-M-MDSCs) expressed higher levels of genes with immunosuppressive functions than other immunosuppressive cell types. Additionally, among the immunosuppressive cell types assessed, M-MDSCs were most significantly enriched in GC tissues relative to adjacent normal tissues. The M-MDSCs in GC tissues expressed significantly higher levels of these markers than adjacent normal tissues; moreover, their presence was most strongly associated with a poor prognosis among the immunosuppressive cells. Immediate early response 3 (IER3), which we identified as a differentially expressed gene between M-MDSCs of GC and adjacent normal tissues, was an independent poor prognostic factor in GC patients (P=0.0003). IER3+ M-MDSCs expressed higher levels of genes with immunosuppressive functions than IER3－ M-MDSCs and were more abundant in treatment-resistant than -responsive GC patie
nts.
Conclusions
The present study suggests that TI-M-MDSCs, especially IER3+ ones, may play a predominant role in the development of the immunosuppressive and ICI-resistant GC TIME.

DOI： 10.1007/s10120-023-01456-4

Publisher：Cancer Science  

Patients with colorectal cancer (CRC) following renal transplantation require long-term immunosuppressants to prevent graft rejection. However, the impact of these immunosuppressants on the tumor immune microenvironment and the roles of immune cells within it remain poorly understood. We conducted comprehensive single-cell RNA sequencing on tumor and normal tissues from four CRC patients post renal transplantation and compared these with published data from 23 non-transplant CRC patients. We set four groups for detailed comparative analysis based on the renal transplantation status and tissue origin: non-renal transplantation normal (nRT_Normal), non-renal transplantation tumor (nRT_Tumor), renal transplantation normal (RT_Normal), renal transplantation tumor (RT_Tumor). Our analysis revealed significant tumor immune microenvironment landscape alterations in the transplantation group. CD8+effector T cells of RT_Tumor showed significantly diminished cytotoxicity and tumor neoantigen recognition (p < 0.0001), while CD4+FOXP3 regulatory T cells of RT_Tumor displayed a higher inhibitory score (p < 0.05), indicating preserved immunomodulatory potential compared with non-transplant CRC. Notably, significantly increased CTLA4 expression in T cells of RT_Tumor was found and testified (p < 0.05). Our findings provide novel mechanistic insights for understanding the immune landscape in renal transplant recipients with CRC and pave the way for potential immunotherapeutic strategies that may improve survival and quality of life for this patient population.

DOI： 10.1111/cas.16409

Scopus

Language：Japanese   Publisher：Japanese Society for Medical and Biological Engineering  

<p>近年，がん微小環境の主要な構成要素である，がん関連線維芽細胞（Cancer-associated fibroblast: CAF）は，がんの悪性化に影響を及ぼすことが明らかになりつつある．近年の研究により，成長因子の分泌などによる生化学的作用や，CAF自身の収縮力による細胞外マトリックスのリモデリングなどの物理的作用が報告されている．このため，がん細胞の浸潤や遊走が影響を受ける．したがって，CAFの作用を詳細に理解することは喫緊の課題となっている．本研究ではCAFの物理的作用が，がん細胞への影響を明らかにすることを考えた．ここで，実際のCAFを用いてしまうと，CAFの物理的作用と生化学的作用が同時に生じてしまい，CAFの物理的作用のみによる，がん細胞への影響を明らかにすることができない．そこで，CAF自身の収縮変形に着目し，CAFと同様に収縮することで物理的作用のみを与えることのできる模擬CAF粒子の開発を試みた．この方法は，3次元で細胞スケールの物理的作用を，がん細胞に与えることができ，両者の物理的相互関係を明らかにできると考えた．本研究の目的は，CAFを模擬した局所的強制変位場を生成する技術の開発である．本研究は，局所的強制変位を負荷することが可能な模擬CAF粒子を作製し，模擬CAF粒子がECM構造にもたらす影響を評価した．</p>

DOI： 10.11239/jsmbe.annual62.147_2

CiNii Research

Publisher：Process Integration and Optimization for Sustainability  

The coordinated integration of agricultural and industrial processes in plant-derived production can offer a solution toward sustainability. However, it is hard for general practitioners to realize the coordinated integration of these processes just based on the precedent fact. A special form clarifying the functions of the required activities should be shared among the practitioners for deliberate system design. In this study, a function model for coordinated integration was developed using the type-zero language of integrated definition for object-oriented design. Inputs, outputs, controls, and mechanisms for the required activities and the relationship between them were analyzed through modeling, after which the model was verified based on actual historical facts in the Japanese cane sugar industry. Finally, as a case study from a different industry, the applicability and limitation of the function model in the palm oil industry are discussed. Although the validity of the model should be confirmed through accumulating future case studies, the structure of the function model should be common to industrial crop-derived productions.

DOI： 10.1007/s41660-023-00337-8

Scopus

Publisher：British Journal of Cancer  

Background: Tertiary lymphoid structures (TLSs) are associated with a favorable prognosis in several cancers. However, the correlation between TLSs and outcomes of esophageal squamous cell carcinoma (ESCC) and the impact of TLSs on the tumor immune microenvironment (TIME) remain unknown. Methods: We pathologically evaluated the significance of TLSs in ESCC focusing on TLS maturation using 180 ESCC specimens and performed single-cell RNA sequencing (scRNA-seq) using 14 ESCC tissues to investigate functional differences of immune cells according to TLS presence. Results: TLS+ cases had better recurrence-free-survival (RFS) (p < 0.0001) and overall survival (OS) (p = 0.0016) compared with TLS- cases. Additionally, mature TLS+ cases had better RFS and OS compared with immature TLS+ cases (p = 0.019 and p = 0.015) and TLS- cases (p < 0.0001 and p = 0.0002). The scRNA-seq showed that CD8+ T cells in TLS+ tumors expressed high levels of cytotoxic signatures and antigen-presentation of dendritic cells (DCs) was enhanced in TLS+ tumors. Immunohistochemistry showed that the densities of tumor-infiltrating CD8+ T cells and DCs were significantly higher in TLS+ tumors than those in TLS- tumors. Conclusions: These data suggest the prognostic and functional significance of TLSs in ESCC and provides new insights into TLSs on the TIME.

DOI： 10.1038/s41416-023-02396-7

Scopus

Publisher：Journal of Cancer Research and Clinical Oncology  

Purpose: This study aimed to demonstrate the involvement of angiogenesis in cancer-associated acinar-to-ductal metaplasia (CA-ADM) lesion of invasive front pancreatic ductal adenocarcinoma (PDAC) and investigate the possible mechanism. Methods: Tissue samples from 128 patients with PDAC and 36 LSL-KrasG12D/+; LSL-Trp53R172H/+; Pdx-1-Cre mice were analyzed. Immunohistochemical assay was performed using HE, anti-CK19 and anti-amylase to confirm the presence of CA-ADM lesions, using anti-CD34 and anti-CD31 to measure microvessel density (MVD), and using anti-CD68, anti-CD163, anti-iNOS, or anti-MMP9 to evaluate the immune microenvironment. We performed multiplex immunohistochemical assay to detect the co-expression of MMP9 and CD68 on macrophage. We examined clinical outcomes and other clinicopathological factors to determine the significance of high-level MVD of CA-ADM on survival and liver metastasis. We performed tube formation assay to evaluate the effect of macrophage on angiogenic capacity in vitro. Results: Angiogenesis was significantly abundant in CA-ADM lesions compared with that in PDAC lesions in human and mouse tissues. High-level MVD in CA-ADM lesions was an independent predictor of poor prognosis (P = 0.0047) and the recurrence of liver metastasis (P = 0.0027). More CD68-positive and CD163-positive macrophages were detected in CA-ADM lesions than in PDAC. The percentage of CD68-positive macrophages was positively correlated with MVD in CA-ADM lesions. Multiplex-immunostaining revealed that MMP9 was expressed in CD68-positive macrophages of CA-ADM lesions. In CA-ADM lesions, the percentage of macrophages was positively correlated with MMP9 expression, which positively correlated with microvessel density. Conclusion: CA-ADM related angiogenesis is a promising predictive marker for poor prognosis of PDAC and may provide an attractive therapeutic target for PDAC.

DOI： 10.1007/s00432-022-04554-5

Scopus

Language：English  

DOI： 10.1186/s40792-023-01699-4

Web of Science

PubMed

Language：English   Publisher：Springer Berlin Heidelberg  

症例は60歳男性。嘔吐を主訴とした。腎盂尿管癌(cT4N0M1 Stage IV)と腹膜播種の既往があり、胃十二指腸吻合術を受けた。腹部CTで胃十二指腸の著明な拡張を認めた。消化管X線造影検査で胃拡張を認め、胃十二指腸吻合による輸出脚に造影剤は届かなかった。胃十二指腸吻合部の狭窄を疑った。ところが上部消化管内視鏡検査では異常なく、内視鏡は輸出脚を通過することから、狭窄は消化管外部からの圧迫によるものと思われた。腎盂尿管癌の腹膜播種による胃十二指腸吻合部狭窄と診断し、保存的治療を施したが、効果がなかった。消化管狭窄に対する内視鏡的拡張およびステント留置を計画した。狭窄部の形状が軟性で、ステント迷入の危険があるため、内視鏡的ステント留置と腹腔鏡下ステント固定の併用術を施行した。全身麻酔下に狭窄部を介して自己拡張型ステントを胃内に留置した。ポート挿入後に腹腔内を観察したところ、結節性病変が胃十二指腸吻合部に癒着していた。腹腔鏡下にステントを輸出脚に縫合固定した。術後にX線造影検査で狭窄部の拡張とステントの位置を確認した。17日後に合併症もなく退院となり、再診時にステント迷入はなかった。

Language：English  

DOI： 10.1186/s40792-023-01671-2

Web of Science

PubMed

Language：English   Publisher：Springer Berlin Heidelberg  

症例は59歳男性。血便を主訴とした。腫瘍マーカー検査で異常所見はみられなかった。大腸内視鏡検査でS字結腸に隆起性病変を認め、生検で腺癌と診断した。腹部造影CTおよび3次元CT血管造影で下腸間膜動脈の分岐走行異常を認めた。下腸間膜動脈は第2腰椎の高さで上腸間膜動脈根部から36mm末梢の位置から分岐していた。その分岐の27mm末梢から中結腸動脈が分岐しており、左結腸動脈とS字結腸動脈はいずれも下腸間膜動脈から分岐していた。腹部大動脈から分岐する動脈はなかった。PET-CTで下腸間膜動脈根部リンパ節にFDGの集積はみられなかった。MRIで肝転移を認めた。以上の所見から下腸間膜動脈の分岐異常を伴うS字結腸癌(cT4aN2aM1a cStage IVA)と術前診断し、腹腔鏡下低位前方切除術を施行した。腹腔内を観察すると、下腸間膜動脈は腹部大動脈と並行して走行しており、腰内臓神経からの結腸枝の神経束を認めた。腫瘍はS字結腸にあり、小腸へ浸潤していた。腸管傍リンパ節を一括郭清し、低位前方切除術を施行した。2ヵ月後に肝転移を切除した。1年半後の再診で再発はなかった。

Language：English  

Background: Tumour immune microenvironment is related with carcinogenesis and efficacy of immunotherapy. B cells play major roles in humoral immunity, but detailed functions of tumour-infiltrating B lymphocytes (TIL-Bs) are unknown. Therefore, our aim was to investigate the functional heterogeneity of TIL-Bs in oesophageal squamous cell carcinoma (ESCC) and lymph nodes (LNs) during chemotherapy.

Methods: Single-cell transcriptome analysis was performed on 23 specimens. We also performed immunohistochemical analysis of immunoglobulin κ C (IGKC), an antibody-secreting cell (ASC) marker, in 166 ESCC samples and evaluated the implication of IGKC in 2-year recurrence free survival (RFS) and 3-year overall survival (OS).

Results: A total of 81,246 cells were grouped into 24 clusters. We extracted B cell clusters based on canonical markers and identified 12 TIL-B subtypes in ESCC. We found that several functions, such as co-stimulation and CD40 signalling, were enhanced in TIL-Bs after chemotherapy. The proportion of naive B cells (NBCs) decreased and B cell activation genes were up-regulated in NBCs after chemotherapy. The proportion of ASCs in tumours increased with the loss of migratory abilities and antibody production in ASCs was promoted after chemotherapy. Differentially expressed genes up-regulated with chemotherapy in ASCs correlated with prolonged survival with oesophageal cancer (p = .028). In a metastatic LN, the ASC proportion increased and B cell differentiation was enhanced. In immunohistochemical analysis, RFS and OS of high IGKC expression cases were significantly better than those of low IGKC expression cases (RFS: p < .0001, OS: p < .0001). And in multivariable analysis, the expression of IGKC was an independent favourable prognostic factor for RFS (hazard ratio (HR): 0.23, 95% confidence interval (CI): 0.12-0.45, p < .0001) and OS (HR: 0.20, 95% CI: 0.086-0.47, p = .0002) in ESCC.

Conclusions: Our findings provide novel insights for the heterogeneity of TIL-Bs during chemotherapy and will be useful to understand the clinical importance of TIL-Bs.

Keywords: neoadjuvant chemotherapy; oesophageal squamous cell carcinoma; single-cell transcriptome; tumour immune microenvironment; tumour infiltrating B lymphocytes.

DOI： 10.1002/ctm2.1181.

Language：English  

Purpose: To determine whether N3 nodal involvement predicts outcomes and whether its prognostic implications vary with tumor location in patients with Stage III colon cancer (CC).

Methods: We defined N3 as lymph node metastases near the bases of the major feeding arteries. We retrospectively examined recurrence rates and patterns by tumor location and sites of lymph node metastases in 57 patients with N3 CC who had undergone curative resections between January 2000 and March 2019. Survival analysis was performed to compare the prognoses of patients with and without N3 lymph node metastasis.

Results: Most N3 patients had large tumors (T ? 3); five had T2 disease. Recurrence occurred quickly in one patient with T2N3M0 disease. Multivariate survival analysis demonstrated that N3 lymph node metastasis is an independent predictor of poor prognosis in Stage III CC patients (P < 0.001). Categorizing N3 patients according to UICC-TNM staging system does not stratify risk of recurrence (P = 0.970). To investigate the impact of tumor location on recurrence risk, we classified N3 CC into two subtypes according to tumor location: metastasis at the base of the superior mesenteric artery in right-sided CC and inferior mesenteric artery in left-sided CC. The former was found to have a statistically significant poorer prognosis than the latter (P = 0.091).

Conclusion: N3 is a robust prognostic marker in CC patients. Recurrence risk varies by tumor location. N3 right-sided CCs with lymph node metastasis at the base of the superior mesenteric artery have poorer prognoses than do N3 left-sided CCs.

Keywords: Feeding artery; Inferior mesenteric artery; N3 colon cancer; Superior mesenteric artery; Tumor sidedness.

DOI： 10.1007/s00423-023-02778-8

Language：English  

DOI： 10.1007/s00423-023-02807-6

Language：English  

DOI： 10.1016/j.isci.2023.106480.

Language：Japanese   Publisher：The Japanese Society of Gastroenterological Surgery  

<p>A 61-year-old woman with a complaint of anemia presented with a flat elevated lesion located in the middle jejunum accompanied by an ulcer at the top of the lesion on oral double-balloon endoscopy. She underwent laparoscopic resection of the jejunum. Pathological findings showed inflammatory cell infiltration and partially lobulated and mildly ectatic capillaries, indicating pyogenic granuloma. Iron deficiency anemia was improved after the operation. Pyogenic granuloma is granulomatous hemangioma that is generally seen in the skin and oral cavity, but rarely in the digestive tract. Pyogenic granuloma can cause obscure gastrointestinal bleeding because it is hemorrhagic.</p>

DOI： 10.5833/jjgs.2021.0085

CiNii Research

Language：Japanese   Publisher：(一社)日本消化器外科学会  

症例は61歳の女性で,貧血の精査で行った経口ダブルバルーン内視鏡で空腸中部に頂部に潰瘍を伴う無茎性の隆起性病変を認めた.黒色便,貧血の改善のため腹腔鏡下小腸部分切除を施行した.病理診断で炎症細胞浸潤および一部小葉状の毛細血管増生の所見を認め,pyogenic granulomaと診断した.術後は貧血の改善を認めた.Pyogenic granulomaは皮膚や口腔内に好発する肉芽組織型血管腫であり,消化管での発生はまれである.易出血性であり消化管出血の原因となりうることから原因不明の消化管出血は本症が原因であることがある.(著者抄録)

Language：Japanese   Publisher：The Japanese Society of Gastroenterological Surgery  

<p>A 69-year-old woman underwent surgery for esophagogastric junction cancer. Multiple subcapsular nodules of several millimeters in size were identified on the liver surface and diagnosed as liver metastases based on pathological examination. Staging laparoscopy was performed after 2 courses of chemotherapy and the same nodules were detected, but the pathological diagnosis was biliary microhamartoma. Finally, the patient underwent curative surgery. In this case, thermal effects of the ultrasonic energy device with an inadequate resection margin in the initial partial hepatectomy was a major obstacle to pathological diagnosis. In addition, the primary lesion was a well differentiated tumor without severe atypia, which could increase the difficulty in pathological diagnosis. This rare entity should be listed as a differential diagnosis of small nodules of the liver, especially when coexisting with gastrointestinal cancers. This report suggests the importance of obtaining a sufficient amount of liver specimens without thermal artifacts, since such artifacts may cause inappropriate identification of structural and morphological findings of tissue components, leading to diagnostic pitfalls.</p>

DOI： 10.5833/jjgs.2021.0101

CiNii Research

Language：Japanese   Publisher：(一社)日本消化器外科学会  

症例は69歳の女性で,食道胃接合部癌に対する手術時に,肝被膜下に5mm弱の白色結節が散在していた.術中迅速診断で腺癌と診断されたため,手術適応外と判断した.化学療法を施行し,その後の審査腹腔鏡時に再度切除した組織で胆管過誤腫と診断され,後日改めて根治術を施行した.本症例は,初回手術時に採取した組織片が小さく,超音波凝固切開装置による熱変性による細胞の変形が,病理診断の大きな障害になったと考えられる.加えて,原発の食道胃接合部癌が高分化型であり,異型が比較的弱い病変であったことも一因といえる.消化器癌に併存した肝腫瘍の診断において,当疾患の存在も念頭におき,微小な病変であってもその組織構築や細胞形態の維持が不十分となり病理診断に影響しないように,アーチファクトが加わらない美麗な標本を十分量採取するよう心がけるべきである.(著者抄録)

Language：English  

For stage II and III esophageal squamous cell carcinoma (ESCC), neoadjuvant chemotherapy (NAC) followed by esophagectomy is recommended in the Japanese guidelines for the diagnosis and treatment of esophageal cancer. However, recurrence of ESCC is common regardless of the NAC regimen and surgical method, and NAC demonstrates limited efficacy against recurrence. Therefore, the present study was conducted to identify risk factors of recurrence of ESCC with surgery after NAC. The outcomes of 51 patients who underwent esophagectomy for ESCC after NAC from 2010 to 2017 at Kyushu University Hospital were retrospectively analyzed. A total of 52 patients with ESCC without NAC followed by esophagectomy from 2001 to 2017 were selected for comparison. Among patients who underwent NAC followed by surgery, only lymphatic invasion (LY; hazard ratio, 2.761; 95% CI, 1.86-6.43, P=0.018) was an independent factor significantly associated with 3-year recurrence-free survival in the multivariate analysis. In patients with pathologic lymph node metastasis (pN) and no LY after NAC, there was significantly less recurrence compared with patients with pN and LY (P=0.0085), whereas in patients without LY after NAC, the presence of pN was not significantly associated with recurrence (P=0.2401). There were significantly fewer LY (+) patients in the NAC (+) group (P=0.0158) compared with those in the NAC (-) group. The presence of LY was an independent risk factor for recurrence of ESCC after esophagectomy following NAC. Overall, adjuvant treatment after surgery may be required in cases with remnant LY after NAC.
Keywords: esophageal squamous cell carcinoma; lymphatic invasion; neoadjuvant chemotherapy; recurrence factor.

DOI： 10.3892/ol.2022.13457

Language：English  

Abstract
Background: Pancreatic ductal adenocarcinoma (PDAC) is characterized by abundant stroma in which microenvironmental (niche) factors promote PDAC progression. In mouse models, reduction of the stroma increased the proportion of poorly differentiated PDAC with a worse prognosis. Here, we aimed to clarify the effects of stroma on PDAC that may define the PDAC phenotype and induce distinct therapeutic responses.
Methods: The molecular features of PDAC based on differentiation grade were clarified by genome and transcriptome analysis using PDAC organoids (PDOs). We identified the dependency on niche factors that might regulate the differentiation grade. A three-dimensional co-culture model with cancer-associated fibroblasts (CAFs) was generated to determine whether CAFs provide niche factors essential for differentiated PDAC. PDOs were subtyped based on niche factor dependency, and the therapeutic responses for each subtype were compared.
Results: The expression profiles of PDOs differed depending on the differentiation grade. Consistent with the distinct profiles, well differentiated types showed high niche dependency, while poorly differentiated types showed low niche dependency. The three-dimensional co-culture model revealed that well differentiated PDOs were strongly dependent on CAFs for growth, and moderately differentiated PDOs showed plasticity to change morphology depending on CAFs. Differentiated PDOs upregulated the expression of mevalonate pathway-related genes correlated with the niche dependency and were more sensitive to simvastatin than poorly differentiated PDOs.
Conclusions: Our findings suggest that CAFs maintain the differentiated PDAC phenotype through secreting niche factors and induce distinct drug responses. These results may lead to the development of novel subtype-based therapeutic strategies.
Keywords: Cancer-associated fibroblast; Organoid; Statin; Stroma-targeting therapy; Subtype-based therapy; Tumor differentiation; Tumor microenvironment.

DOI： 10.1186/s13046-022-02301-9

Language：English  

Abstract
Background/objectives: Pancreatic ductal adenocarcinoma (PDAC) is characterized by excessive desmoplasia and autophagy-dependent tumorigenic growth. Pancreatic stellate cells (PSCs) as a predominant stromal cell type play a critical role in PDAC biology. We have previously reported that autophagy facilitates PSC activation, however, the mechanism remains unknown. We investigated the mechanism of autophagy in PSC activation.
Methods: We compared gene expression profiles between patient-derived PSCs from pancreatic cancer and chronic pancreatitis using a microarray. The stromal expression of target gene in specimen of PDAC patients (n = 63) was analyzed. The effect of target gene on autophagy and activation of PSCs was investigated by small interfering RNAs transfection, and the relationship between autophagy and ER stress was investigated. We analyzed the growth and fibrosis of xenografted tumor by orthotopic models.
Results: In analysis of gene expression microarray, endoplasmic reticulum aminopeptidase 2 (ERAP2) upregulated in cancer-associated PSCs was identified as the target gene. High stromal ERAP2 expression is associated with a poor prognosis of PDAC patients. Knockdown of ERAP2 inhibited unfolded protein response mediated autophagy, and led to inactivation of PSCs, thereby attenuating tumor-stromal interactions by inhibiting production of IL-6 and fibronectin. In vivo, the promoting effect of PSCs on xenografted tumor growth and fibrosis was inhibited by ERAP2 knockdown.
Conclusions: Our findings demonstrate a novel mechanism of PSCs activation regulated by autophagy. ERAP2 as a promising therapeutic target may provide a novel strategy for the treatment of PDAC.
Keywords: Endoplasmic reticulum stress; Stromal remodeling; Tumor-stromal interaction; Tunicamycin.

DOI： 10.1016/j.pan.2021.09.012

Language：English  

Abstract
Purpose: Preoperative sarcopenia worsens postoperative outcomes in various cancer types including colorectal cancer. However, we often experienced postoperative anastomotic leakage in muscular male patients such as Judo players, especially in rectal cancer surgery with lower anastomosis. It is controversial whether the whole skeletal muscle mass impacts the potential for anastomotic failure in male rectal cancer patients. Thus, the purpose of this study was to clarify whether skeletal muscle mass impacts anastomotic leakage in rectal cancer in men.
Methods: We reviewed the medical charts of male patients suffering from rectal cancer who underwent colo-procto anastomosis below the peritoneal reflection without a protective diverting stoma. We measured the psoas muscle area and calculated the psoas muscle index.
Results: One hundred ninety-seven male rectal cancer patients were enrolled in this study. The psoas muscle index was significantly higher in patients with anastomotic leakage (P<0.001). Receiver operating characteristic curve determined the optimal cut-off value of the psoas muscle index for predicting anastomotic leakage as 812.67 cm2/m2 (sensitivity of 60% and specificity of 74.3%). Multivariate analysis revealed that high psoas muscle index (risk ratio [RR], 3.933; P<0.001; 95% confidence interval [CI], 1.917-8.070) and super low anastomosis (RR, 2.792; P=0.015; 95% CI, 1.221-6.384) were independent predictive factors of anastomotic leakage.
Conclusion: This study showed that male rectal cancer patients with a large psoas muscle mass who underwent lower anastomosis had a higher rate of postoperative anastomotic leakage.
Keywords: Anastomotic leak; Lower colo-anorectal anastomosis; Psoas muscle index; ROC curve; Rectal neoplasms

DOI： 10.3393/ac.2022.00122.0017

Publisher：GCCE 2022 - 2022 IEEE 11th Global Conference on Consumer Electronics  

In recent years, computer-aided diagnosis based on deep learning concepts has become an attractive research topic in medical imaging. Most of these works utilized supervised learning methods that required prior pathological knowledge. However, it is necessary to extract potential features in images, based on unsupervised learning in order to obtain new pathological findings. For this reason, we implemented unsupervised cluster analysis based on maximization of mutual information to classify pathological images of pancreatic cancer into discrete categories.

DOI： 10.1109/GCCE56475.2022.10014057

Scopus

Language：English  

Abstract
Pancreatic ductal adenocarcinoma (PDAC) is characterized by notorious resistance to current therapies attributed to inherent tumor heterogeneity and highly desmoplastic and immunosuppressive tumor microenvironment (TME). Unique proline isomerase Pin1 regulates multiple cancer pathways, but its role in the TME and cancer immunotherapy is unknown. Here, we find that Pin1 is overexpressed both in cancer cells and cancer-associated fibroblasts (CAFs) and correlates with poor survival in PDAC patients. Targeting Pin1 using clinically available drugs induces complete elimination or sustained remissions of aggressive PDAC by synergizing with anti-PD-1 and gemcitabine in diverse model systems. Mechanistically, Pin1 drives the desmoplastic and immunosuppressive TME by acting on CAFs and induces lysosomal degradation of the PD-1 ligand PD-L1 and the gemcitabine transporter ENT1 in cancer cells, besides activating multiple cancer pathways. Thus, Pin1 inhibition simultaneously blocks multiple cancer pathways, disrupts the desmoplastic and immunosuppressive TME, and upregulates PD-L1 and ENT1, rendering PDAC eradicable by immunochemotherapy.
Keywords: Pin1; cancer immune evasion; cancer-associated fibroblasts; chemotherapy; combination therapy; immuni checkpoint therapy; pancreatic cancer; targeted therapy; tumor immune microenvironment; tumor microenvironment.

DOI： 10.1016/j.cell.2021.07.020

Language：English  

Abstract
Introduction: The proportion of patients aged 75 years and over who undergo minimally invasive surgery for gastric cancer is increasing. However, the safety and feasibility of laparoscopic gastrectomy (LG) in this age group is controversial. This study aimed to evaluate whether LG is safe and effective in patients aged 75 years and over.
Methods: The study included 728 patients with early and advanced gastric cancer who underwent curative LG between 2009 and 2017; 166 of these patients (22.8%) were aged 75 or over. All surgeries were performed laparoscopically. Selected clinical factors were compared between the 166 patients aged 75 years and over and the 562 patients aged under 75 years.
Results: There were significant differences in presence of comorbidity, respiratory function and American Society of Anesthesiologists physical status scores between the older and younger groups. The older patients more frequently developed complications than the younger ones, particularly postoperative pneumonia. According to multivariate analyses of all participants, age, chronic obstructive pulmonary disease (COPD), and D2 lymphadenectomy were independent risk factors for postoperative pneumonia. Advanced stage and D2 lymphadenectomy were independent risk factors in the older group, whereas only COPD was an independent risk factor in the younger group.
Conclusions: LG for gastric cancer can be safely performed in patients aged over 75 years with an acceptable complication rate. However, the present data suggest that care should be taken in selecting LG with D2 lymphadenectomy to treat advanced cancer in these patients because the risk of postoperative complications, especially postoperative pneumonia, increases.

DOI： 10.1111/ases.12883

Language：English  

Abstract
Pancreatic cancer cells (PCCs) are surrounded by an abundant stroma, which is produced by pancreatic stellate cells (PSCs). PSCs promote tumor cell proliferation and invasion. The objective of the current study was to identify compounds that suppress PSC activation. Gene expression profiles of cancer-derived fibroblasts and normal fibroblasts were used, and the pathway analysis suggested altered pathways that were chosen for validation. It was found that the 'neuroactive ligand-receptor interaction' pathway from the Kyoto Encyclopedia of Genes and Genomes pathway analysis was one of the altered pathways. Several compounds related with this pathway were chosen, and changes in PSC activity were investigated using fluorescence staining of lipid droplets, reverse transcription-quantitative PCR, western blotting, and invasion and migration assays. Among these candidates, duloxetine, a serotonin-noradrenaline reuptake inhibitor, was found to suppress PSC activation and disrupt tumor-stromal interaction. Thus, duloxetine may be a potential drug for suppressing PSC activation and pancreatic cancer growth.
Keywords: drug repositioning; duloxetine; pancreatic cancer; pancreatic stellate cells; tumor microenvironment.

DOI： 10.3892/ol.2021.13005

Language：English  

Abstract
Background/objectives: Pancreatic stellate cells (PSCs) are involved in abundant desmoplasia, which promotes cancer cell aggressiveness and resistance to anti-cancer drugs. Therefore, PSCs are suggested to be a promising therapeutic target by attenuating PSC activation to inhibit tumor-stromal interactions with pancreatic cancer cells. Here, we developed a screen to identify compounds that reduce the activity of PSCs and investigated the effect of candidates on pancreatic cancer.
Methods: Lipid droplet accumulation in PSCs was used to observe differences in PSC activity and a new high-throughput screening platform that quantified lipid droplets in PSCs was established. A library of 3398 Food and Drug Administration-approved drugs was screened by this platform. Validation assays were performed in vitro and in vivo.
Results: Thirty-two compounds were finally selected as candidate compounds by screening. These compounds decreased α-smooth muscle actin expression and inhibited autophagic flux in PSCs in vitro. Among the candidates, three drugs selected for validation assays inhibited the proliferation and migration of PSCs and invasion of cancer cells by disrupting tumor-stromal interactions. Production of extracellular matrix molecules was also decreased significantly by this treatment. In vivo testing in xenograft models showed that dopamine antagonist zuclopenthixol suppressed tumor growth; this suppression was significantly increased when combined with gemcitabine.
Conclusions: A new screening platform that focused on the morphological features of PSCs was developed. Candidate drugs from this screening suppressed PSC activation and tumor growth. This screening system may be useful to discover new compounds that attenuate PSC activation.
Keywords: Cell-based screen; Dopamine antagonist; Pancreatic cancer; Pancreatic stellate cell; Stromal targeting therapy.

DOI： 10.1016/j.pan.2021.04.002

Language：English  

Background: Pancreatic stellate cells (PSCs) occupy the majority of the pancreatic cancer microenvironment, contributing to aggressive behavior of pancreatic cancer cells (PCCs). Recently, anti-fibrotic agents have proven to be an effective strategy against cancer, but clinical trials have shown little efficacy, and the driving mechanism remains unknown. N-acetyl-cysteine (NAC) is often used for pulmonary cystic fibrosis. Pioglitazone, an agonist of peroxisome proliferator-activated receptor gamma, was habitually used for type II diabetes, but recently reported to inhibit metastasis of PCCs. However, few studies have focused on the effects of these two agents on cancer-stromal interactions.
Method: We evaluated the expression of α-smooth muscle actin (α-SMA) and the number of lipid droplets in PSCs cultured with or without NAC. We also evaluated changes in invasiveness, viability, and oxidative level in PSCs and PCCs after NAC treatment. Using an indirect co-culture system, we investigated changes in viability, invasiveness, and migration of PSCs and PCCs. Combined treatment effects of NAC and Pioglitazone were evaluated in PSCs and PCCs. In vivo, we co-transplanted KPC-derived organoids and PSCs to evaluate the effects of NAC and Pioglitazone's combination therapy on subcutaneous tumor formation and splenic xenografted mouse models.
Results: In vitro, NAC inhibited the viability, invasiveness, and migration of PSCs at a low concentration, but not those of PCCs. NAC treatment significantly reduced oxidative stress level and expression of α-SMA, collagen type I in PSCs, which apparently present a quiescent-like state with a high number of lipid droplets. Co-cultured PSCs and PCCs mutually promoted the viability, invasiveness, and migration of each other. However, these promotion effects were attenuated by NAC treatment. Pioglitazone maintained the NAC-induced quiescent-like state of PSCs, which were reactivated by PCC-supernatant, and enhanced chemosensitivity of PCCs. In vivo, NAC and Pioglitazone's combination suppressed tumor growth and liver metastasis with fewer stromal components and oxidative stress level.
Conclusion: NAC suppressed activated PSCs and attenuated cancer-stromal interactions. NAC induces quiescent-like PSCs that were maintained in this state by pioglitazone treatment.

DOI： 10.1186/s13046-021-01939-1

Language：English  

Purpose: T1 gastric cancer (GC) with seven or more metastatic lymph nodes is extremely rare, and very few clinical studies have been conducted to evaluate the clinicopathological features of their recurrence.

Methods: We retrospectively analyzed the outcomes of T1 GC and T2-4 GC patients who had multiple nodal metastases after radical surgery from 2006 to 2020. Propensity score matching was performed to compare the two groups of patients.

Results: After propensity score matching, 18 of 22 patients in the T1 group and 36 of 144 patients in the T2-4 group were selected. Recurrence occurred in six patients (33.3%) in the T1 group. In the T1 group, the most common site of initial recurrence was bone (15.0%). The prevalence of bone recurrence was significantly higher in the T1 group than in the T2-4 group (P = 0.02). The median interval time between radical surgery and bone recurrence was 24 months, and the median survival time after bone recurrence was 14 months.

Conclusion: Bone recurrence was more frequently identified as an initial recurrence site in T1 GC cases with multiple metastases after radical surgery compared with that in T2-4 GC cases. Careful attention should be paid to postoperative bone recurrence in the long-term postoperative course of these patients.

DOI： 10.1007/s00423-021-02231-8

Language：English  

Abstract
Pancreatic stellate cells (PSCs) play a key role in desmoplastic stroma, which is a characteristic of pancreatic ductal adenocarcinoma (PDAC), and they also enhance the malignancy of pancreatic cancer cells. Our previous study reported chloroquine's mitigating effects on PSC activation; however, the drug is known to induce adverse effects in clinical practice. The present study aimed to reduce chloroquine doses and develop a useful pre-treatment that targets PSCs using nanoparticles. Poly lactic-co-glycolic acid (PLGA) nanoparticles were used as carriers and loaded with indocyanine green (Nano-ICG) or chloroquine (Nano-CQ). Tumor accumulation of Nano-ICG was evaluated using an in vivo imaging system. The effects of chloroquine, Nano-CQ and/or chemotherapy drug gemcitabine were investigated in an orthotopic xenograft mouse model. Nano-ICG selectively accumulated in pancreatic tumors and persisted therein for over 7 days after administration. Additionally, Nano-ICG accumulated in the peritoneal metastasized regions, but not in the liver, kidney and normal pancreatic tissues. Nano-CQ reduced the density of activated PSCs at lower chloroquine doses and significantly restrained tumor progression in combination with gemcitabine. In conclusion, the PLGA nanosystem successfully delivered the drug to pancreatic tumors. Nano-CQ efficiently reduced PSC activation and may be a promising novel pre-treatment strategy for PDAC.
Keywords: chloroquine; combination therapy; desmoplasia; nanoparticles; orthotopic xenograft; pancreatic cancer; pancreatic ductal adenocarcinoma; pre-treatment; stroma.

DOI： 10.3892/ol.2021.12894

Language：English  

Abstract
Purpose: The aim of this study was to investigate the genetic mutation profiles of Japanese pancreatic ductal adenocarcinoma (PDAC) patients.
Methods: Next-generation sequencing was performed using FoundationOne® CDx on 17 PDAC patients who were treated by surgical resection at Kyushu University Hospital between February 2016 and January 2019. The tumor mutational burden and microsatellite instability status were also assessed.
Results: There were 16 patients (94%) with KRAS mutations, 13 (76%) with TP53 mutations, three (18%) with SMAD4 mutations, and one (6%) with a CDKN2A mutation. All patients had at least one pathogenic variant or a likely pathogenic variant. No patient had targeted therapies that matched with any clinical benefit according to FoundationOne® CDx. An unresectable PDAC patient with BRCA2-mutant disease was successfully treated by conversion surgery using platinum-based neoadjuvant chemotherapy.
Conclusions: Currently, FoundationOne® CDx might be difficult to use on PDAC patients, although further investigations with larger study populations are called for.
Keywords: FoundationOne; Gene profiling; Next-generation sequencing; Pancreatic cancer; Pancreatic ductal adenocarcinoma.

DOI： 10.1007/s00595-020-02123-2

Language：English  

DOI： 10.1371/journal.pone.0228015

Language：English  

Abstract
Purpose: To identify the incidence of extraction site incisional hernia following gastrectomy for gastric cancer and its significant risk factors, including the subcutaneous fat area.
Methods: We reviewed data gathered prospectively on patients with gastric cancer, who underwent gastrectomy between 2008 and 2012 at Kyushu University Hospital, Fukuoka, Japan. The subcutaneous fat area (SFA) and visceral fat area (VFA) were measured using axial computed tomography at the level of the L4 and L3 transverse processes, and the L2-L3 intervertebral disc. The primary endpoint of the rate of extraction site incisional hernia was based on the computed tomography and clinical data including hospital follow-up reports.
Results: After applying the inclusion and exclusion criteria, 320 patients were included in this retrospective analysis: 3.1% (10/320) had extraction site incisional hernias after a mean follow-up of 11 months. Multivariate analysis revealed that age and the SFA were independent risk factors (age ≥ 70.5 years: P = .013, odds ratio: 9.116, 95% confidence interval 1.581-52.553; L4 SFA ≥ 124 cm2: P = .004, odds ratio: 13.752, 95% confidence interval 2.290-82.582).
Conclusion: Age and the SFA were independent risk factors for extraction site incisional hernia in patients undergoing gastrectomy for gastric cancer.

DOI： 10.1007/s00595-020-02039-x

Language：English  

Abstract
Introduction: The proportion of patients aged 75 years and over who undergo minimally invasive surgery for gastric cancer is increasing. However, the safety and feasibility of laparoscopic gastrectomy (LG) in this age group is controversial. This study aimed to evaluate whether LG is safe and effective in patients aged 75 years and over.
Methods: The study included 728 patients with early and advanced gastric cancer who underwent curative LG between 2009 and 2017; 166 of these patients (22.8%) were aged 75 or over. All surgeries were performed laparoscopically. Selected clinical factors were compared between the 166 patients aged 75 years and over and the 562 patients aged under 75 years.
Results: There were significant differences in presence of comorbidity, respiratory function and American Society of Anesthesiologists physical status scores between the older and younger groups. The older patients more frequently developed complications than the younger ones, particularly postoperative pneumonia. According to multivariate analyses of all participants, age, chronic obstructive pulmonary disease (COPD), and D2 lymphadenectomy were independent risk factors for postoperative pneumonia. Advanced stage and D2 lymphadenectomy were independent risk factors in the older group, whereas only COPD was an independent risk factor in the younger group.
Conclusions: LG for gastric cancer can be safely performed in patients aged over 75 years with an acceptable complication rate. However, the present data suggest that care should be taken in selecting LG with D2 lymphadenectomy to treat advanced cancer in these patients because the risk of postoperative complications, especially postoperative pneumonia, increases.

DOI： 10.1111/ases.12883

Language：English  

Abstract:Progress in diagnostic modalities, surgical procedures, and multidisciplinary treatment for pancreatic diseases has increased the number of long-term survivors after pancreatic resection. Several reports have focused on high-risk lesions (HRLs), including high-grade pancreatic intraepithelial neoplasia (PanIN), pancreatic ductal adenocarcinoma, high-grade intraductal papillary mucinous neoplasm (IPMN), and IPMN with an associated invasive carcinoma, in the remnant pancreas after partial pancreatic resection for pancreatic cancer or IPMN. The etiology of HRLs in the remnant pancreas is thought to be either isolated local recurrence of the initial lesion in the remnant pancreas or a newly developed primary lesion. Although it is difficult to distinguish between local recurrence and a new primary lesion, comparison of genetic alterations between two lesions may help with this distinction. Early detection of HRLs in the remnant pancreas may improve the prognosis of patients, and several investigators have proposed predictive factors for HRLs in the remnant pancreas after partial pancreatic resection for pancreatic cancer or IPMN. The reported short- and long-term outcomes of surgical resection of HRLs in the remnant pancreas are relatively favorable. Life-long surveillance of the remnant pancreas is recommended after partial pancreatic resection for pancreatic cancer or IPMN.

DOI： 10.1007/s00595-019-01852-3

Language：English  

Abstract
Cancer‑associated fibroblasts (CAFs) promote the progression of pancreatic ductal adenocarcinoma (PDAC) via tumor‑stromal interactions. Neutrophil extracellular traps (NETs) are extracellular DNA meshworks released from neutrophils together with proteolytic enzymes against foreign pathogens. Emerging studies suggest their contribution to liver metastasis in several types of cancer. Herein, in order to investigate the role of NETs in liver metastasis in PDAC, the effects of NET inhibitors on spontaneous PDAC mouse models were evaluated. It was demonstrated that DNase I, a NET inhibitor, suppressed liver metastasis. For further investigation, further attention was paid to liver micrometastasis and an experimental liver metastasis mouse model was used that was generated by intrasplenic tumor injection. Furthermore, DNase I also suppressed liver micrometastasis and notably, CAFs accumulated in metastatic foci were significantly decreased in number. In vitro experiments revealed that pancreatic cancer cells induced NET formation and consequently NETs enhanced the migration of hepatic stellate cells, which was the possible origin of CAFs in liver metastasis. On the whole, these results suggest that NETs promote liver micrometastasis in PDAC via the activation of CAFs.

DOI： 10.3892/ijo.2019.4951

Language：English  

Abstract
Lymph node metastasis is an independent prognostic factor in pancreatic cancer. However, the mechanisms of lymph node colonization are unknown. As a mechanism of lymphatic metastasis, it has been reported for other types of cancer that spheroids from tumor cells cause circular chemorepellent‑induced defects (CCIDs) in lymphatic endothelial monolayers. In pancreatic cancer, such mechanisms of metastasis have not been elucidated. The present study evaluated the involvement of this new mechanism of metastasis in pancreatic cancer and investigated the associated factors. In human pancreatic cancer tissue, it was observed that clusters of cancer cells penetrated the wall of lymphatic ducts around the primary tumor. An in vitro co‑culture system was then used to analyze the mechanisms of tumor cell‑mediated disruption of lymphatic vessels. Time‑lapse microscopic imaging revealed that spheroids from pancreatic cancer cells caused circular defects in lymphatic endothelial monolayers. CCID formation ability differed depending on the cell line. Neither aggregation of spheroids nor adhesion to lymphatic endothelial cells (LECs) exhibited a significant correlation with this phenomenon. The addition of supernatant from cultured cancer cells enhanced CCID formation. Microarray analysis revealed that the expression of S100 calcium binding protein P (S100P) was significantly increased when LECs were treated with supernatant from cultured cancer cells. Addition of a S100P antagonist significantly suppressed the migration of LECs and CCID formation. The present findings demonstrated that spheroids from pancreatic cancer cells caused circular defects in lymphatic endothelial monolayers. These CCIDs in pancreatic cancer were partly regulated by S100P, suggesting that S100P may be a promising target to inhibit lymph node metastasis.

DOI： 10.3892/ijo.2019.4812

Language：English  

Abstract
BACKGROUND:Extracellular signal-regulated kinases (ERKs) have been related to multiple cancers, including breast cancer, hepatocellular cancer, lung cancer and colorectal cancer. ERK1/2 inhibitor can suppress growth of KRAS-mutant pancreatic tumors by targeting cancer cell. However, no studies have shown the expression of ERK1/2 on pancreatic stromal and its effect on pancreatic cancer-stromal interaction.
METHODS:Immunohistochemistry and western blotting were performed to detect the expression of p-ERK1/2 in pancreatic tissues and cells. Cell viability assay was used to study IC50 of ERK inhibitor on pancreatic cancer cells (PCCs) and primary cancer-associated pancreatic stellate cells (PSCs). Transwell migration, invasion, cell viability assay, senescence β-galactosidase staining were performed to determine the effect of ERK inhibitor on PCCs and PSCs in vitro and in vivo. The expression of key factors involved in autophagy and epithelial-to-mesenchymal transition (EMT) process were evaluated by western blotting. The expression of key factors related to cell invasiveness and malignancy were confirmed by qRT-PCR. Co-transplantation of PCC Organoid and PSC using a splenic xenograft mouse model was used to evaluated combined treatment of ERK inhibitor and autophagy inhibitor.
RESULTS:Immunohistochemical staining in pancreatic tumor samples and transgenetic mice detected p-ERK1/2 expression in both cancer cells and stromal cells. In pancreatic tissues, p-ERK1/2 was strongly expressed in cancer-associated PSCs compared with cancer cells and normal PSCs. PSCs were also significantly more sensitive to ERK1/2 inhibitor treatment. Inhibition of ERK1/2 suppressed EMT transition in HMPCCs, upregulated cellular senescence markers, activated autophagy in cancer-associated PSCs; and suppressed cancer-stromal interaction, which enhanced invasiveness and viability of cancer cells. We also found that chloroquine, an autophagy inhibitor, suppressed ERK inhibition-induced autophagy and promoted PSC cellular senescence, leading to significantly decreased cell proliferation. The combination of an ERK inhibitor and autophagy inhibitor suppressed liver metastasis in a splenic pancreatic cancer organoid xenograft mouse model.
CONCLUSIONS: These data indicate that inhibition of ERK1/2 in cancer-associated pancreatic stellate cells suppresses cancer-stromal interaction and metastasis.

DOI： 10.1186/s13046-019-1226-8

Language：English  

Abstract:Progress in diagnostic modalities, surgical procedures, and multidisciplinary treatment for pancreatic diseases has increased the number of long-term survivors after pancreatic resection. Several reports have focused on high-risk lesions (HRLs), including high-grade pancreatic intraepithelial neoplasia (PanIN), pancreatic ductal adenocarcinoma, high-grade intraductal papillary mucinous neoplasm (IPMN), and IPMN with an associated invasive carcinoma, in the remnant pancreas after partial pancreatic resection for pancreatic cancer or IPMN. The etiology of HRLs in the remnant pancreas is thought to be either isolated local recurrence of the initial lesion in the remnant pancreas or a newly developed primary lesion. Although it is difficult to distinguish between local recurrence and a new primary lesion, comparison of genetic alterations between two lesions may help with this distinction. Early detection of HRLs in the remnant pancreas may improve the prognosis of patients, and several investigators have proposed predictive factors for HRLs in the remnant pancreas after partial pancreatic resection for pancreatic cancer or IPMN. The reported short- and long-term outcomes of surgical resection of HRLs in the remnant pancreas are relatively favorable. Life-long surveillance of the remnant pancreas is recommended after partial pancreatic resection for pancreatic cancer or IPMN.

DOI： 10.1007/s00595-019-01852-3

Language：English  

Abstract
BACKGROUND: It is difficult to determine whether a second high-risk lesion, including pancreatic ductal adenocarcinoma or high-grade pancreatic intraepithelial neoplasm, is a metachronous multifocal lesion or represents local recurrence after resection of the first high-risk lesion. This study attempts to clarify the characteristics of second high-risk lesions in the remnant pancreas using genetic analyses.
METHODS: Clinicopathologic data were collected from 12 patients who underwent pancreatectomy for a second high-risk lesion in the remnant pancreas. We performed mutational and immunohistochemical analyses of 4 major genes-KRAS, TP53, CDKN2A, and SMAD4-associated with pancreatic ductal adenocarcinoma progression, as well as targeted next-generation sequencing.
RESULTS: Mutations in the four genes in the second high-risk lesion were consistent with the first lesion in four patients but were inconsistent in the remaining eight patients, and thus we considered that the latter eight patients likely had metachronous multifocal high-risk lesions and the other four patients had local recurrence. The estimated cumulative recurrence rate after resection of the second high-risk lesion was greater in the local recurrence group compared with the metachronous multifocal group, and the estimated cumulative disease-specific survival rate was greater in the metachronous multifocal group. Targeted next-generation sequencing demonstrated that the second lesions in the metachronous multifocal high-risk lesion group showed differences in founder mutations compared with the first lesion. In the local recurrence group, the founder mutations in the second lesion were common with those in the first lesion.
CONCLUSION: Genetic assessment might help discriminate metachronous multifocal high-risk lesions from local recurrence.

DOI： 10.1016/j.surg.2018.10.025

Language：English  

Abstract
PURPOSE:This study aimed at investigating the function and significance of CD110 expression in pancreatic cancer.
METHODS:We performed immunohistochemical staining for CD110 expression in tumor samples from 86 patients with pancreatic cancer. We evaluated clinical outcomes and other clinicopathological factors to determine the significance of CD110 on survival and liver metastasis. We examine thrombopoietin-CD110 signaling in cancer cell extravasation in vitro and in vivo. We investigated the effects of CD110 knockdown on liver metastasis in a splenic xenograft mouse model.
RESULTS:CD110 expression in cancer cells was associated with low-histological-grade invasive ductal carcinoma, and patients with high CD110 expression had poorer prognosis (P = 0.0003). High CD110 expression was an independent predictor of liver metastasis (P = 0.0422). Knockdown of CD110 expression significantly attenuated cell migration and invasion. Treatment with thrombopoietin promoted pancreatic cancer cell extravasation. In the presence of thrombopoietin, CD110 increased cell viability through the activation of the ERK-MYC signaling pathway. Knockdown of CD110 expression inhibited liver metastases in the mouse model.

DOI： 10.1007/s00432-019-02860-z

Language：English  

Abstract
The pancreas is an organ prone to inflammation, fibrosis, and atrophy because of an abundance of acinar cells that produce digestive enzymes. A characteristic of pancreatic cancer is the presence of desmoplasia, inflammatory cell infiltration, and cancer-associated acinar atrophy (CAA) within the invasive front. CAA is characterized by a high frequency of small ducts and resembles acinar-to-ductal metaplasia (ADM). However, the clinical significance of changes in acinar morphology, such as ADM with acinar atrophy, within the tumor microenvironment remains unclear. Here, we find that ADM within the invasive front of tumors is associated with cell invasion and desmoplasia in an orthotopic mouse model of pancreatic cancer. An analysis of resected human tumors revealed that regions of cancer-associated ADM were positive for TGFα, and that this TGFα expression was associated with primary tumor size and shorter survival times. Gene expression analysis identified distinct phenotypic profiles for cancer-associated ADM, sporadic ADM and chronic pancreatitis ADM. These findings suggest that the mechanisms driving ADM differ according to the specific tissue microenvironment and that cancer-associated ADM and acinar atrophy contribute to tumor cell invasion of the local pancreatic parenchyma.

DOI： 10.1016/j.canlet.2018.12.005

Language：English  

Abstract
BACKGROUND/AIM:
Metabolomics is widely used for biomarker discovery, but conventional mass-spectrometry extraction procedures lose the spatial localization of metabolites. In this study, we directly analyzed breast carcinoma tissues embedded in frozen tissue microarrays (fTMAs) using MALDI mass-spectrometry imaging (MALDI-MSI).
MATERIALS AND METHODS:
A total of 119 breast tissues (84 carcinoma and 35 normal) were used. MSI data were extracted from each tissue.
RESULTS:
Overall, 185 of 1,915 peaks which were commonly detected in 60% of target areas were subjected to further analysis. One hundred and fifty-two peaks of carcinoma showed significantly higher intensity than normal. Comparing metabolite profiles from carcinoma and normal tissues, energy charge (EC) and the sum of adenosine phosphate compound (AXP) indicated significantly higher intensities in cancerous tissues than normal. But comparisons of EC and AXP among lymph node metastasis, tumor size and tumor subtypes indicated no significant differences.
CONCLUSION:
Breast carcinoma tissues had higher EC and AXP values than normal. MALDI-MSI could be a tool for characterizing breast carcinoma.

DOI： doi:10.21873/anticanres.12723

Language：English  

DOI： 10.1016/j.biomaterials.2017.10.029.

Language：English  

DOI： 10.1245/s10434-018-6761-6

Language：English  

Abstract
It is important to estimate the postmortem interval in forensic autopsy. Many methods to estimate the postmortem interval have been reported, and are typically associated with internal examination. However, there are issues such as rejection of autopsy by the family and a lack of forensic doctor in internal examination. Therefore, it is necessary to develop new methods, such as autopsy imaging, that can substitute for internal examination. Here, we first evaluated whether gas volume in the body increased with postmortem interval. Time-dependent X-ray CT imaging of euthanized Crl:CD (SD) rats (n = 3) was performed immediately after euthanasia and at seven subsequent time points up to 168 h (7 days) at 24-hour intervals. The data revealed that gas volume in the body increased in a time-dependent manner. Next, we reconstructed 3D images of isolated gas and calculated the gas volume using Amira software. In all cases, the volume of both portal venous gas and intestinal gas increased in a time-dependent manner. The volume of portal venous gas increased exponentially, while the volume of intestinal gas increased in a linearly with time. These data might be suggested that the postmortem gas volume change is one of indicators for estimating the postmortem interval. In addition, it would be possible to estimate more accurate postmortem interval by combining not only gas volume changes at the above two sites but also gas volume changes of the other sites such as heart cavities, kidney parenchyma, or abdominal aorta.

DOI： 10.1016/j.legalmed.2017.12.010

Language：English  

Specific cell populations leading the local invasion of cancer are called “leading cells”. However, the underlying mechanisms are unclear. Here, we identified leading cells in pancreatic cancer and deter- mined how these cells lead and promote cancer cell invasion in the extracellular matrix (ECM). Using three-dimensional matrix remodeling assay, we found that pancreatic stellate cells (PSCs) frequently invaded the collagen matrix with pancreatic cancer cells (PCCs), which invaded behind the invading PSCs. In addition, invading PSCs changed the alignment of collagen fibers, resulting in ECM remodeling and an increase in the parallel fibers along the direction of invading PSCs. Endo180 expression was higher in PSCs than in PCCs, Endo180 knockdown in PSCs attenuated the invasive abilities of PSCs and co- cultured PCCs, and decreased the expression level of phosphorylated myosin light chain 2 (MLC2). In mouse models, Endo180-knockdown PSCs
suppressed tumor growth and changes in collagen fiber orientation in co-transplantation with PCCs. Our findings suggest that PSCs lead the local invasion of PCCs by physically remodeling the ECM, possibly via the function of Endo180, which reconstructs the actin cell skeleton by phosphorylation of MLC2.

DOI： 10.1016/j.canlet.2017.10.010

Language：English  

Abstract
BACKGROUND: It is difficult to determine whether a second high-risk lesion, including pancreatic ductal adenocarcinoma or high-grade pancreatic intraepithelial neoplasm, is a metachronous multifocal lesion or represents local recurrence after resection of the first high-risk lesion. This study attempts to clarify the characteristics of second high-risk lesions in the remnant pancreas using genetic analyses.
METHODS: Clinicopathologic data were collected from 12 patients who underwent pancreatectomy for a second high-risk lesion in the remnant pancreas. We performed mutational and immunohistochemical analyses of 4 major genes-KRAS, TP53, CDKN2A, and SMAD4-associated with pancreatic ductal adenocarcinoma progression, as well as targeted next-generation sequencing.
RESULTS: Mutations in the four genes in the second high-risk lesion were consistent with the first lesion in four patients but were inconsistent in the remaining eight patients, and thus we considered that the latter eight patients likely had metachronous multifocal high-risk lesions and the other four patients had local recurrence. The estimated cumulative recurrence rate after resection of the second high-risk lesion was greater in the local recurrence group compared with the metachronous multifocal group, and the estimated cumulative disease-specific survival rate was greater in the metachronous multifocal group. Targeted next-generation sequencing demonstrated that the second lesions in the metachronous multifocal high-risk lesion group showed differences in founder mutations compared with the first lesion. In the local recurrence group, the founder mutations in the second lesion were common with those in the first lesion.
CONCLUSION: Genetic assessment might help discriminate metachronous multifocal high-risk lesions from local recurrence.

DOI： 10.1016/j.surg.2018.10.025

Language：English  

Abstract
The aim of this study was to assess the feasibility of prophylactic pancreatojejunostomy after enucleation or limited pancreatic resection regarding the risk of postoperative pancreatic fistula (PF). We retrospectively reviewed the medical records of 32 patients who underwent enucleation or limited pancreatic resection and compared the clinical parameters between patients with (n = 10) and without (n = 22) prophylactic pancreatojejunostomy. Prophylactic pancreatojejunostomy was performed in patients with a possible high risk ofPF. No operation-related mortality occurred. Operation time was significantly longer (P < 0.01) and blood loss significantly greater (P < 0.01) in patients with pancreatojejunostomy. Overall complications were more frequent (P = 0.02) and postoperative hospital stay was significantly longer (P = 0.02) in patients with pancreatojejunostomy. However, other assessed factors including the prevalence of postoperative PF did not differ between groups. In conclusion, prophylactic pancreatojejunostomy is feasible, and its efficacy in preventing PF after enucleation or limited pancreatic resection in high-risk patients will require further study.

Language：English  

Abstract
Disturbed balance between immune surveillance and tolerance may lead to poor clinical outcomes in some malignancies. In paired analyses of adenocarcinoma and normal mucosa from 142 patients, we found a significant increase of the CD4/CD8 ratio and accumulation of regulatory T cells (Tregs) within the adenocarcinoma. The increased frequency of Tregs correlated with the local infiltration and extension of the tumor. There was concurrent maturation arrest, upregulation of programmed death-1 expression, and functional impairment in CD8+ T cells (CTLs) isolated from the adenocarcinoma. Adenocarcinoma-associated Tregs directly inhibit the function of normal human CTLs in vitro. With histopathological analysis, Foxp3+ Tregs were preferentially located in stroma. Concurrent transcriptome analysis of epithelial cells, stromal cells, and T cell subsets obtained from carcinomatous and normal intestinal samples from patients revealed a distinct gene expression signature in colorectal adenocarcinoma-associated Tregs, with overexpression of CCR1, CCR8, and TNFRSF9, whereas their ligands CCL4 and TNFSF9 were found upregulated in cancerous epithelium. Overexpression of WNT2 and CADM1, associated with carcinogenesis and metastasis, in cancer-associated stromal cells suggests that both cancer cells and stromal cells play important roles in the development and progression of colorectal cancer through the formation of a tumor microenvironment. The identification of CTL anergy by Tregs and the unique gene expression signature of human Tregs and stromal cells in colorectal cancer patients may facilitate the development of new therapeutics against malignancies.

DOI： 10.4049/jimmunol.1701222

Language：English  

Abstract
BACKGROUND AND AIMS:
Colonic endoscopic submucosal dissection (ESD) is time-consuming and bears a high risk of perforation. The aim of the present study was to compare the safety and efficacy between novel articulating devices and conventional ESD in live porcine colon models.
METHODS:
Thirty ESDs in ten pigs were carried out at three different locations (15, 25, and 35 cm from the anus) by the conventional method (n = 15) and by the new method (n = 15). Procedure times, adverse events (perforation, bleeding), and damage to the muscular layer were recorded, and the ESD time per unit area of the specimens was calculated.
RESULTS:
The perforation rate using the conventional method was 6.7% (1/15), whereas that using the new method was 0.0%. The number of sites of muscular damage was significantly lower in the new than conventional method (6 vs. 37, respectively; P = 0.024). The mean procedure time was significantly shorter in the new than conventional method (4.6 ± 2.0 vs. 7.0 ± 4.1 min/cm2, respectively; P = 0.042).
CONCLUSIONS:
Use of the new ESD method allows for reduced adverse events and a shortened resection time.

DOI： 10.1007/s00464-018-6408-5.

Language：English  

Stroma invasion is an important step in pancreatic cancer progression. However, how pancreatic ductal adenocarcinoma (PDAC) with ductal structure invades the surrounding stroma has not been clear. Here, we elucidated the mechanism of stromal invasion of PDAC, using organoids. From resected PDAC specimens, we established human PDAC organoids, which developed ductal and basement membrane (BM) structures. When the organoids were co-cultured with pancreatic stellate cells (PSCs) in a collagen matrix, organoids lost their BM and ductal structures, and invaded collagen matrix more frequently than did mono-cultured organoids. Interestingly, direct contact by PSCs to PDAC organoids was observed before BM destruction. Matrix metalloproteinase (MMP) 2 or membrane type-1 MMP (MT1MMP) knockdown in PSCs significantly attenuated BM destruction by PSCs, and retained the ductal structures in organoids. Our results imply that direct contact by PSCs induces BM destruct
ion and stromal invasion of PDAC via MMP2 which binds to MT1MMP on PSCs.

DOI： 10.1016/j.canlet.2018.03.031

Language：English  

Abstract
BACKGROUND:
Two ligation techniques can be applied in laparoscopy for left-sided colorectal cancer: (1) high-tie (HT), transection at the level of the inferior mesenteric artery (IMA); and (2) low-tie (LT), transection below the IMA, at the level of superior rectal artery (SRA), preserving the left colic artery (LCA). However, even with preoperative images, it can still be a challenge to identify these structures due to intraoperative individual conditions. In this study, we assess the use intraoperative ultrasonography (IOUS) to aid us in identifying the IMA and its branches to the SRA, LCA, and sigmoid artery.
METHODS:
We performed IOUS in 18 patients diagnosed with left-sided colorectal cancer. Preoperatively, a three-dimensional computed tomography (3D-CT) angiography was obtained in majority of the patients, to visualize the IMA and its branches. Two patients were contraindicated to receive a contrast study, hence, was unable to undergo 3D-CT angiography. The resected specimen was grossly examined for the study. The bifurcation types were identified and compared using different modalities: preoperative 3D-CT, IOUS, and gross examination of the resected specimen.
RESULTS:
The branching of the IMA revealed by IOUS was consistent to the findings preoperatively by the 3D-CT and postoperatively by the resected specimen. The IOUS result of the two patients without preoperative 3D-CT evaluation was also consistent with the post-operative bifurcation type.
CONCLUSIONS:
IOUS is an easy and feasible modality which aids in detecting the branching of the IMA during LT and HT ligation in laparoscopic left-sided colorectal surgery. It can serve as an adjunct modality for 3D-CT angiography and can also be considered a safe alternative option for cases wherein 3D-CT angiography is unavailable

DOI： 10.1007/s00464-018-6302-1

Language：English  

Abstract
Although recent studies revealed that adipose tissue accelerates pancreatic tumor progression with excessive extracellular matrix, key players for desmoplasia in the adipose microenvironment remains unknown. Here, we investigated the roles of adipose tissue-derived stromal cells (ASCs) in desmoplastic lesions and tumor progression by in vitro and in vivo experiments. In a three-dimensional (3-D) organotypic fat invasion model using visceral fat from CAG-EGFP mice, GFP-positive fibroblastic cells infiltrated toward cancer cells. When tumor cells were inoculated into transplanted visceral fat pads in vivo, tumor weights and stromal components were enhanced compared to subcutaneous and orthotopic tumor cells inoculated without fat pads. Expression of αSMA in established human ASCs was lower compared to cancer associated fibroblasts, and the 3-D collagen matrices produced by ASCs cultured in cancer cell-conditioned medium changed from loose to dense structures that affected the motility of cancer cells. Microarray analyses revealed upregulation of S100A4 in ASCs, while S100A4-positive stromal cells were observed at extrapancreatic invasion sites of human pancreatic cancer. The present findings indicate that ASCs are recruited to extrapancreatic invasion sites and produce dense collagen matrices that lead to enhanced tumor progression. Both inhibition of ASCs recruitment and activation could lead to a novel antistromal therapy.

DOI： 10.1002/ijc.31775

Language：English  

DOI： 10.1007/s00464-018-6188-y

Language：English  

Abstract
BACKGROUND:
Difficulties in endoscopic operations and therapeutic procedures seem to occur due to the complexity of operating the endoscope dial as well as difficulty in performing synchronized movements with both hands. We developed a prototype robotic-assisted flexible endoscope that can be controlled with a single hand in order to simplify the operation of the endoscope. The aim of this study was to confirm the operability of the robotic-assisted flexible endoscope (RAFE) by performing endoscopic submucosal dissection (ESD).
METHODS:
Study 1: ESD was performed manually or with RAFE by an expert endoscopist in ex vivo porcine stomachs; six operations manually and six were performed with RAFE. The procedure time per unit circumferential length/area was calculated, and the results were statistically analyzed. Study 2: We evaluated how smoothly a non-endoscopist can move a RAFE compared to a manual endoscope by assessing the designated movement of the endoscope.
RESULTS:
Study 1: En bloc resection was achieved by ESD using the RAFE. The procedure time was gradually shortened with increasing experience, and the procedure time of ESD performed with the RAFE was not significantly different from that of ESD performed with a manual endoscope. Study 2: The time for the designated movement of the endoscope was significantly shorter with a RAFE than that with a manual endoscope as for a non-endoscopist.
CONCLUSIONS:
The RAFE that we developed enabled an expert endoscopist to perform the ESD procedure without any problems and allowed a non-endoscopist to control the endoscope more easily and quickly than a manual endoscope. The RAFE is expected to undergo further development.

DOI： 10.1007/s00464-018-6188-y.

Language：Japanese  

Abstract：＜ポイント＞腹腔鏡下膵手術を保険診療として実施する場合には,厚生労働省や学会が示す適応と指針を熟知する必要がある.腹腔鏡下膵頭十二指腸切除術の保険診療下での運用に際しては,施設認定,学会との連携,National Clinical Database(NCD)への登録を行うことが義務付けられており,学会連携申請制度と,NCD術前登録制度が整備されている.保険診療として定められていない腹腔鏡下膵手術を実施する場合には,当該厚生局と内議のうえ保険診療準用の是非を確認するか,公費負担もしくは自費診療としなければならない.いずれの場合も自施設倫理委員会の認可を受けることが必須である.(著者抄録)

Language：English  

Abstract
OBJECTIVES:
The development of an effective screening method for pancreatic ductal adenocarcinoma (PDAC) is of paramount importance. This study assessed the diagnostic utility in pancreatic diseases of duodenal markers during upper gastrointestinal endoscopy (GIE) or endoscopic ultrasonography.

METHODS:
This study prospectively enrolled 299 consecutive participants, including 94 patients with PDACs, 144 patients with other pancreatic diseases, and 61 normal individuals as control subjects. All subjects underwent upper GIE or endoscopic ultrasonography either at Kyushu University Hospital (Fukuoka, Japan) or the Mayo Clinic (Jacksonville, Fla) from October 2011 to July 2014. Duodenal fluid (DF) was collected without secretin stimulation and of carcinoembryonic antigen and S100 calcium-binding protein P (S100P) concentrations were measured.

RESULTS:
Concentrations of S100P in DF were significantly higher in patients with PDAC and chronic pancreatitis than in control subjects (P < 0.01). A logistic regression model that included age found that the sensitivity and specificity of S100P concentration in diagnosing stages 0/IA/IB/IIA PDAC were 85% and 77%, respectively, with an area under the receiver operating characteristic curve of 0.82. Carcinoembryonic antigen concentrations in DF of patients with pancreatic disease did not differ significantly from control subjects.

CONCLUSIONS:
Analysis of S100P concentration in DF, in combination with routine screening upper GIE, may facilitate the detection of PDAC.

DOI： 10.1097/MPA.0000000000000940

Language：English  

Abstract
PURPOSE:
We developed a touchless display system that allows the user to control the medical imaging software via hand gestures in the air. We conducted this study to verify the effectiveness of this novel touchless display system as a tool for assisting with surgical imaging.
METHODS:
The patient's computed tomography (CT) data are generally observed on a display during surgery. The "Dr. aeroTAP" touchless display system was developed to generate virtual mouse events based on the position of one hand. We conducted comparative analyses of using the Dr. aeroTAP vs. using a regular mouse (control group) by measuring the time to select a 3D image from 24 thumbnail images on a screen (study 1) and to then see the CT image on the DICOM viewer (study 2).
RESULTS:
We used the Dr. aeroTAP in 31 hepato-biliary operative procedures performed at our hospital. In study 1, which measured the time required to select one of 24 thumbnails, there were significant differences between the mouse and Dr. aeroTAP groups for all five surgeons who participated (P < 0.001). In study 2, there were also significant differences in the time required for CT DICOM images to be displayed (P < 0.001).
CONCLUSIONS:
The touchless interface proved efficient for allowing the observation of surgical images while maintaining a sterile field during surgery.

DOI： 10.1007/s00595-017-1541-7

Language：English  

Abstract
BACKGROUND:
We previously reported the use of an inverted T-shaped method to obtain a suitable view for hand sewing to close the common entry hole when the linear stapler was fired for esophagojejunostomy after laparoscopic total gastrectomy (LTG). This conventional method involved insertion of the fixed cartridge fork to the Roux limb and the fine movable anvil fork to the esophagus to avoid perforation of the jejunum. However, insertion of the movable anvil fork to the esophagus during this procedure often requires us to strongly push down the main body of the stapler with the fixed cartridge fork to bring the direction of the anvil fork in line with the direction of the long axis of the esophagus while controlling the opening of the movable anvil fork. We therefore modified this complicated inverted T-shaped method using a linear stapler with a movable cartridge fork. This modified method involved insertion of the movable cartridge fork into the Roux limb followed by natural, easy insertion of the fixed anvil fork into the esophagus without controlling the opening of the movable cartridge fork.
METHODS:
We performed LTG in a total of 155 consecutive patients with gastric cancer from November 2007 to December 2015 in Kyushu University Hospital. After LTG, we performed the conventional inverted T-shaped method using a linear stapler with a fixed cartridge fork in 61 patients from November 2007 to July 2011 (fixed cartridge group). From August 2011, we used a linear stapler with a movable cartridge fork and performed the modified inverted T-shaped method in 94 patients (movable cartridge group). We herein compare the short-term outcomes in 94 cases of LTG using the modified method (movable cartridge fork) with those in 61 cases using the conventional method (fixed cartridge fork).
RESULTS:
We found no significant differences in the perioperative or postoperative events between the movable and fixed cartridge groups. One case of anastomotic leakage occurred in the fixed cartridge group, but no anastomotic leakage occurred in the movable cartridge group.
CONCLUSIONS:
Although there were no remarkable differences in the short-term outcomes between the movable and fixed cartridge groups, we believe that the modified inverted T-shaped method is technically more feasible and reliable than the conventional method and will contribute to the improved safety of LTG.

Language：English  

Abstract
Pancreatic cancer progression involves components of the tumor microenvironment, including stellate cells, immune cells, endothelial cells, and the extracellular matrix. Although peripancreatic fat is the main stromal component involved in extra-pancreatic invasion, its roles in local invasion and metastasis of pancreatic cancer remain unclear. This study investigated the role of adipose tissue in pancreatic cancer progression using genetically engineered mice (Pdx1-Cre; LSL-KrasG12D; Trp53R172H/+) and an in vitro model of organotypic fat invasion. Mice fed a high fat diet had significantly larger primary pancreatic tumors and a significantly higher rate of distant organ metastasis than mice fed a standard diet. In the organotypic fat invasion model, pancreatic cancer cell clusters were smaller and more elongated in shape and showed increased fibrosis. Adipose tissue-derived conditioned medium enhanced pancreatic cancer cell invasiveness and gemcitabine resistance, as well as inducing morphologic changes in cancer cells and increasing the numbers of lipid droplets in their cytoplasm. The concentrations of oleic, palmitoleic, and linoleic acids were higher in adipose tissue-derived conditioned medium than in normal medium, with these fatty acids significantly enhancing the migration of cancer cells. Mature adipocytes were smaller and the concentration of fatty acids in the medium higher when these cells were co-cultured with cancer cells. These findings indicate that lipolytic and fibrotic changes in peripancreatic adipose tissue enhance local invasiveness and metastasis via adipocyte-released fatty acids. Inhibition of fatty acid uptake by cancer cells may be a novel therapy targeting interactions between cancer and stromal cells.
KEYWORDS:
adipose microenvironment; extra-pancreatic invasion; fatty acids; lipolysis; pancreatic cancer

DOI： 10.18632/oncotarget.15430.

Language：English  

Abstract
Pancreatic cancer is characterized by increased hyperplasia of fibrotic tissue, termed desmoplasia, and lymph node metastasis is an independent prognostic factor in this disease. However, there are no reports focused on desmoplasia in pancreatic cancer lymph node metastases. The present study evaluated a range of factors and investigated their association with poor prognosis in pancreatic cancer cases with lymph node metastasis, including the degree of desmoplasia in lesions. To identify the poor prognostic factors associated with lymph node metastasis, the present study retrospectively reviewed the clinical data of 65 patients with lymph node metastases that underwent surgical pancreatic cancer resection between 2007 and 2012 at a single institution. The investigation focused on the degree of fibrosis in metastatic lesions in 216 lymph nodes, and investigated associations with prognosis or clinicopathological findings. The ratios of the fibrotic area in metastatic lymph node lesions were evaluated and classified into three categories, high (≥70%), moderate (10-70%) and low (<10%). Desmoplasia was not observed in cancer-free lymph nodes. The size of metastatic lymph node lesions was additionally measured, and a significant association between metastatic lesion size and the degree of desmoplasia was observed (P<0.001). The degree of desmoplasia was additionally associated with local extranodal invasion. In the analysis of 65 pancreatic cancer patients with metastatic lymph nodes, the presence of multiple metastatic lymph nodes with moderate or high desmoplasia was significantly associated with poor survival (high, P=0.0048; moderate/high, P=0.0075). Of several clinicopathological factors, the presence of multiple metastatic lymph nodes with high or moderate desmoplasia was associated with overall survival in univariate (P=0.0098) and multivariate (P=0.0466) analyses. The degree of desmoplasia in metastatic lymph nodes is associated with lesion size, and the presence of multiple metastatic lymph nodes with desmoplasia is an independent poor prognostic factor, suggesting that the desmoplasia may have an important role in the malignant progression of lymph node metastases.
KEYWORDS:
desmoplasia; locally extranodal invasion; lymph node metastasis; pancreatic cancer; prognostic factor

DOI： 10.3892/ol.2017.6549

Language：English  

Abstract
BACKGROUND:
Circumportal pancreas is a rare congenital pancreatic anomaly. The aim of this study was to clarify the clinical characteristics of patients with circumportal pancreases undergoing pancreatectomy.
METHODS:
The medical records of 508 patients who underwent pancreatectomy were retrospectively reviewed. The prevalence of circumportal pancreas and related anatomical variations were assessed. Surgical procedures and postoperative outcomes were compared in patients with and without circumportal pancreas.
RESULTS:
Circumportal pancreas was observed in 9 of the 508 patients (1.7%). In all nine patients, the portal vein was completely encircled by the pancreatic parenchyma above the level of the splenoportal junction, and the main pancreatic duct ran dorsal to the portal vein. The rate of variant hepatic artery did not differ significantly in patients with and without circumportal pancreas. Pancreatic fistula developed more frequently in patients with than without circumportal pancreas (44% vs. 14%, p = 0.03), but other clinical parameters did not differ significantly in these two groups.
CONCLUSIONS:
Despite being rare, circumportal pancreas may increase the risk of postoperative pancreatic fistula in patients undergoing pancreatectomy. However, a prospective, large-cohort study is necessary to determine the real incidence of relevant anatomical variations and the definitive clinical significance of this rare anomaly.

Language：English  

BACKGROUND:The clinical importance of intraoperative peritoneal cytology in patients with pancreatic cancer remains incompletely understood.METHODS:Peritoneal washing samples were collected from 411 consecutive patients with pancreatic ductal adenocarcinoma from 1996 to 2014. Of the 411 patients, 335 underwent macroscopically curative resection and 76 with noncurative factors did not undergo resection. We compared long-term outcomes between patients with positive cytology (cytology+) and those with negative cytology (cytology-) and investigated the importance of clinicopathologic factors.RESULTS:Of 335 patients with curative resection, 300 (89.6%) were cytology- and 35 (10.4%) were cytology+. The median overall survival of cytology+ patients was less than that of cytology- patients (16 vs 31 months, respectively; P &LT; .0001). The median overall survival of cytology+ patients with noncurative factors was significantly worse than that of cytology+ pat
ients with curative resection (6.9 vs 16.0 months, respectively; P = .0023). The median disease-free survival of cytology+ patients was less than that of cytology- patients (6.5 vs 16 months, respectively; P &LT; .0001). In the multivariate analysis, cytology+ was an independent prognostic factor for overall survival and disease-free survival.CONCLUSION:Cytology+ without noncurative factors was a predictive factor for a poor prognosis. Therefore, it is important to regard patients with pancreatic cancer characterized by cytology+ as a special group that may warrant more aggressive adjuvant therapy.

Language：English  

The interaction between the cancer cells and the peritoneal mesothelial cells (PMCs) plays an important role in the peritoneal dissemination in several types of cancer. However, the role of PMCs in the peritoneal dissemination of pancreatic cancer remains unclear. In the present study, we investigated the interaction between the pancreatic cancer cells (PCCs) and the PMCs in the formation of peritoneal dissemination in vitro and in vivo. The tumor-stromal interaction of PCCs and PMCs significantly enhanced their mobility and invasiveness and enhanced the proliferation and anoikis resistance of PCCs. In a 3D organotypic culture model of peritoneal dissemination, co-culture of PCCs and PMCs significantly increased the cells invading into the collagen gel layer compared with mono-culture of PCCs. PMCs pre-invaded into the collagen gel, remodeled collagen fibers, and increased parallel fiber orientation along the direction of cell invasion. In the tissues
of peritoneal dissemination of the KPC (LSL-KrasG12D/+; LSL-Trp53R172H/+;Pdx-1-Cre) transgenic mouse, the monolayer of PMCs was preserved in tumor-free areas, whereas PMCs around the invasive front of peritoneal dissemination proliferated and invaded into the muscle layer. In vivo, intraperitoneal injection of PCCs with PMCs significantly promoted peritoneal dissemination compared with PCCs alone. The present data suggest that the cancer-associated PMCs have important promoting roles in the peritoneal dissemination of PCCs. Therapy targeting cancer-associated PMCs may improve the prognosis of patients with pancreatic cancer.

Language：English  

BACKGROUND & AIMS: Pancreatic stellate cells (PSCs) changefrom a quiescent to activated state in the tumor environmentand secrete extracellular matrix (ECM) molecules and cytokinesto increase the aggressiveness of tumors. However, it is notclear how PSCs are activated to produce these factors, orwhether this process can be inhibited. PSCs have morphologicand functional similarities to hepatic stellate cells, whichundergo autophagy to promote fibrosis and tumor growth. Weinvestigated whether autophagy activates PSCs, which promotesdevelopment of the tumor stroma and growth ofpancreatic tumors in mice. METHODS: We used immunofluorescencemicroscopy and immunohistochemistry to analyzepancreatic tumor specimens from 133 patients who underwentpancreatectomy in Japan from 2000 to 2009. PSCs werecultured from pancreatic tumor tissues or tissues of patientswith chronic pancreatitis; these were analyzed by immunofluorescencemicroscopy, immunoblots, quantitative
reversetranscription polymerase chain reaction, and in assays forinvasiveness, proliferation, and lipid droplets. Autophagy wasinhibited in PSCs by administration of chloroquine or transfectionwith small interfering RNAs. Proteins were knockeddown in immortalized PSCs by expression of small hairpinRNAs. Cells were transplanted into pancreatic tails of nudemice, and tumor growth and metastasis were quantified. RESULTS:Based on immunohistochemical analyses, autophagywas significantly associated with tumor T category (P シ .018),histologic grade (P シ .001), lymph node metastases (P &LT; .001),stage (P シ .009), perilymphatic invasion (P シ .001), and perivascularinvasion (P シ .003). Autophagy of PSCs was associatedwith shorter survival times of patients with pancreatic cancer.PSC expression of microtubule-associated protein 1 lightchain 3, a marker of autophagosomes, was associated with pooroutcomes (shorter survival time, disease recurrence) forpati
ents with pancreatic cancer (relative risk of shorter survivaltime, 1.56). Immunoblots showed that PSCs from pancreatictumor samples expressed higher levels of markers of autophagythan PSCs from chronic pancreatitis samples. Inhibitorsof autophagy increased the number of lipid droplets of PSCs,indicating a quiescent state of PSCs, and reduced their productionof ECM molecules and interleukin 6, as well as theirproliferation and invasiveness in culture. PSCs exposedto autophagy inhibitors formed smaller tumors in nude mice(P シ .001) and fewer liver metastases (P シ .018) with lessperitoneal dissemination (P シ .018) compared to PSCs notexposed to autophagy inhibitors. CONCLUSIONS: AutophagicPSCs produce ECM molecules and interleukin 6 and areassociated with shorter survival times and disease recurrencein patients with pancreatic cancer. Inhibitors of PSC autophagymight reduce pancreatic tumor invasiveness by altering thetumor stroma.

DOI： 10.1053/j.gastro.2017.01.010

Language：English  

Background: Salinomycin has cytotoxic effects on various types of malignancy and induces autophagy.However, it has not been clarified whether autophagy induced by salinomycin treatment has a protectiveor cytotoxic role. We investigated whether salinomycin affects autophagy in pancreatic cancer cells andwhether autophagy induced by salinomycin treatment has a protective or cytotoxic role in these cells.Methods: We investigated the effect of salinomycin using three pancreatic cancer cell lines. We investigatedeffect on proliferation and the CD133 positive fraction using flow cytometry. In addition, wemonitored the change in autophagic activity after salinomycin treatment using fluorescent immunostaining,western blotting, and flow cytometry. Finally, knockdown of ATG5 or ATG7 by siRNA was usedto investigate the impact of autophagy inhibition on sensitivity to salinomycin.Results: Salinomycin suppressed the proliferation of pancreatic cancer cells in a co
ncentration dependentmanner, and reduced the CD133 positive fraction. Salinomycin enhanced autophagy activity in these cellsin a concentration dependent manner. Autophagy inhibition made pancreatic cancer cells more sensitiveto salinomycin.Conclusions: Our data provide the first evidence indicating that autophagy induced by salinomycin havea protective role in pancreatic cancer cells. A new therapeutic strategy of combining salinomycin,autophagy inhibitors, and anticancer drugs could hold promise for pancreatic cancer treatment.

DOI： 10.1016/j.pan.2017.08.009

Language：English  

Desmoplasia contributes to the aggressive behavior of pancreatic cancer. However, recent clinical trials testing several antifibrotic agents on pancreatic cancer have not shown clear efficacy. Therefore, further investigation of desmoplasia-targeting antifibrotic agents by another mechanism is needed. Calpeptin, an inhibitor of calpains, suppressed fibroblast function and inhibited fibrosis. In this study, we investigated the anticancer effects of calpeptin on pancreatic cancer. We investigated whether calpeptin inhibited tumor progression using a mouse xenograft model. We used quantitative RT-PCR to evaluate the expression of calpain-1 and calpain-2 mRNA in pancreatic cancer cells (PCCs) and pancreatic stellate cells (PSCs). We also undertook functional assays, including proliferation, migration, and invasion, to evaluate the inhibitory effects of calpeptin on PCCs and PSCs. Quantitative RT-PCR indicated that PCCs and PSCs expressed calpain-2 mRNA. C
alpeptin reduced tumor volume (P = 0.0473) and tumor weight (P = 0.0471) and inhibited the tumor desmoplastic reaction (P &LT; 0.001) in xenograft tumors in nude mice. Calpeptin also inhibited the biologic functions of PCCs and PSCs including proliferation (P = 0.017), migration (P = 0.027), and invasion (P = 0.035) in vitro. Furthermore, calpeptin reduced the migration of PCCs and PSCs by disrupting the cancer-stromal interaction (P = 0.0002). Our findings indicate that calpeptin is a promising antitumor agent for pancreatic cancer, due not only to its suppressive effect on PCCs and PSCs but also its disruption of the cancer-stromal interaction.

Language：Japanese  

Abstract：2012年改訂版IPMN/MCN国際診療ガイドラインでは、IPMNの手術適応の閾値が引き上げられ、経過観察が選択されることが多くなった。IPMNの認知度が高まり、画像診断技術も進歩し、IPMNは日常診療でも遭遇する頻度の高い疾患となったため、今後も経過観察されるIPMNが増加することが予想される。非切除IPMNの経過観察では、IPMN自体の悪性化だけでなく、IPMNとは別に発生する通常型膵癌にも注意が必要である。IPMNの進行は緩徐なため、短期間での経過観察を必要としない一方で、通常型膵癌の生物学的悪性度を考慮すると、短期間での経過観察を必要とする可能性もある。しかしいまだ至適経過観察方法は確立されていない。現在、日本膵臓学会主導で「分枝型IPMN前向き追跡調査」が進行中であり、この研究結果によりIPMNの自然史が明らかとなり、至適経過観察方法が確立されることが期待される。

Language：English  

Pancreatic stellate cells (PSCs) enhance the malignant behavior of pancreatic cancer by interacting with cancer cells and producing extracellular matrix (ECM). To date, several stroma-targeted therapies for pancreatic cancer have been attempted, but these therapies are still not in practical use. Integrins expressed in stromal cells are involved in fibrosis of several organs, as well as promoting tumor malignancy. We investigated whether CD51, also known as integrin αV, expressed in PSCs was associated with stromal formation of pancreatic cancer and enhancement of tumor malignancy. We also assessed the effects of suppression of CD51 in PSCs on pancreatic cancer. Immunohistochemistry for CD51 in resected pancreatic cancer tissues showed that high expression of CD51 in the tumor stroma was associated with lymph node metastasis (P=0.025), positive pathologic margin (P=0.025), and shorter patient survival times (P=0.043). Lentivirus-mediated short hairpin RNA knockdown of CD51 decreased the proliferation and migration of PSCs. Quantitative real-time polymerase chain reaction showed that expression levels of genes related with ECM and tumor-stromal interactions were decreased by CD51 knockdown in PSCs. In a co-implantation model of pancreatic cancer cells and PSCs, tumor growth in vivo was inhibited by CD51 knockdown in PSCs (P<0.05). We also found reduced tumor stroma and decreased proliferation of cancer cells in implanted cancer tissues with CD51-silenced PSCs (P<0.05). Our results showed that CD51 expression in pancreatic cancer stroma is associated with enhanced tumor malignancy and that CD51 may be a potential therapeutic target for pancreatic cancer.

Language：English  

Abstract

Metastasis is the main cause of cancer-associated death, and metastasis of pancreatic cancer remains difficult to treat because of its aggressiveness. MicroRNAs (miRNAs) play crucial roles in the regulation of various human transcripts, and many miRNAs have been reported to correlate with cancer metastasis. We identified an anti-metastatic miRNA, miR-5100, by investigating differences in miRNA profiling between highly metastatic pancreatic cancer cells and their parental cells. Overexpression of miR-5100 inhibited colony formation (P<0.05), cell migration (P<0.0001) and invasion (P<0.0001) of pancreatic cancer cells. In addition, we identified a possible target of miR-5100, podocalyxin-like 1 (PODXL), and demonstrated miR-5100 directly binds to the 3' untranslated region of PODXL and post-transcriptionally regulates its expression in pancreatic cancer cells. Silencing PODXL resulted in diminished cell migration (P<0.0001) and invasion (P<0.05). We also clarified the close relationship between expression of PODXL in human pancreatic cancer specimens and liver metastasis (P=0.0003), and determined that post-operative survival was longer in the low-PODXL expression group than in the high-PODXL expression group (P<0.05). These results indicate that miR-5100 and PODXL have considerable therapeutic potential for anti-metastatic therapy and could be potential indicators for cancer metastases in patients with pancreatic cancer.

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Desmoplasia and hypoxia in pancreatic cancer mutually affect each other and create a tumor-supportive microenvironment. Here, we show that microenvironment remodeling by hypoxic pancreatic stellate cells (PSCs) promotes cancer cell motility through alteration of extracellular matrix (ECM) fiber architecture. Three-dimensional (3-D) matrices derived from PSCs under hypoxia exhibited highly organized parallel-patterned matrix fibers compared with 3-D matrices derived from PSCs under normoxia, and promoted cancer cell motility by inducing directional migration of cancer cells due to the parallel fiber architecture. Microarray analysis revealed that procollagen-lysine, 2-oxoglutarate 5-dioxygenase 2 (PLOD2) in PSCs was the gene that potentially regulates ECM fiber architecture under hypoxia. Stromal PLOD2 expression in surgical specimens of pancreatic cancer was confirmed by immunohistochemistry. RNA interference-mediated knockdown of PLOD2 in PSCs abrogated para
llel fiber architecture of 3-D matrices, leading to decreased directional migration of cancer cells within the matrices. In conclusion, these findings indicate that hypoxia-induced PLOD2 in PSCs creates a permissive microenvironment for migration of cancer cells through architectural regulation of stromal ECM in pancreatic cancer.

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PURPOSE:Laparoscopic and open surgical skills differ distinctly from one another. Our institute provides laparoscopic surgical skills training for currently active surgeons throughout Japan. This study was performed to evaluate the effectiveness of our 2-day standardized laparoscopic surgical skills training program over its 10-year history.METHODS:We analyzed the data on trainee characteristics, outcomes of skills assessments at the beginning and end of the program, and self-assessment after 6 months using a questionnaire survey.RESULTS:From January 2004 to December 2013, 914 surgeons completed the 2-day training program. Peaks in postgraduate years of experience occurred at years 2, 8, and 17. Suturing and knot tying times were significantly shorter at the end than beginning of the program (p &LT; 0.001). However, the numbers of misplaced and loose sutures, maximum misplacement distance, and number of injuries to the rubber sheet were significantly
higher at the end of the program (p &LT; 0.001). A questionnaire at 6 months post-training revealed significant improvements in the overall skills and forceps manipulation (p &LT; 0.0001) and a significantly shorter mean operation time for laparoscopic cholecystectomy (p &LT; 0.001).CONCLUSION:Our 2-day training program for active Japanese surgeons is thus considered to be effective; however, continued voluntary training is important and further outcomes assessments are needed.

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OBJECTIVES:To clarify the usefulness of molecular biomarkers for distinguishing invasive carcinoma derived from intraductal papillary mucinous neoplasms (IPMNs [Inv-IPMN]) from concomitant pancreatic ductal adenocarcinoma (PDAC).METHODS:Data from 19 patients with resected concomitant PDAC were retrospectively reviewed. KRAS/GNAS mutations and immunohistochemical (IHC) expression of p53 and p16/CDKN2A were assessed in both IPMN and distinct PDAC. As controls, KRAS/GNAS mutations and IHC labeling were assessed between invasive and noninvasive components in 1 lesion of 22 independent patients.RESULTS:KRAS/GNAS mutation status of invasive and noninvasive components in Inv-IPMN was consistent in 18 (86%) of 21 patients. Conversely, mutational patterns in IPMN and distinct PDAC in the same pancreas differed from each other in 17 (89%) of 19. There were 10 (53%) and 8 (42%) of 19 patients who showed the same p53 and p16/CDKN2A staining between concomitant PDAC and d
istinct IPMN. In the Inv-IPMN cohort, 19 (86%) of 22 patients showed the same IHC expression pattern between the noninvasive and invasive components.CONCLUSIONS:It may be possible to distinguish Inv-IPMN from concomitant PDAC by assessing these molecular biomarkers. More precise distinction of Inv-IPMN and concomitant PDAC will lead to adequate recognition of the natural history of IPMNs and hence optimal management.

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BACKGROUND: Radical antegrade modular pancreatosplenectomy (RAMPS) is a modification of standard retrograde pancreatosplenectomy (SRPS) used to achieve the dissection of N1 lymph nodes, early vascular control, and negative posterior margins. However, there have been few comparative studies regarding the clinical outcomes of the RAMPS and SRPS procedures.METHODS: Ninety-three patients underwent distal pancreatectomy for the treatment of pancreas body and tail adenocarcinoma between 2000 and 2014. Clinicopathologic data were retrospectively analyzed in this study. We compared short- and long-term outcomes between RAMPS and SRPS. In addition, we investigated the significance of clinicopathologic factors in left-sided pancreatic cancers.RESULTS: Fifty-three patients underwent RAMPS and 40 patients underwent SRPS. RAMPS revealed a larger number of retrieved lymph nodes [28.4 ± 11.6 vs 20.7 ± 10.1; P = 0.0016], more frequent R0 resection [90.5 vs 67.5
%; P = 0.0053], less intraoperative bleeding than SRPS [485.4 ± 63.3 vs 682.3 ± 72.8 ml; P = 0.0444], and shorter operating time (267.3 ± 11.5 vs 339.4 ± 13.2 min; P &LT; 0.0001) as compared with SRPS. In comparing RAMPS and SRPS, RAMPS showed a tendency for improvement of the median survival times than SRPS (47 vs 34 months) (P = 0.1920). In the multivariate analysis, R1 resection, histologic grade, and vascular invasion for overall survival (OS) were found to be independent factors.CONCLUSIONS: There were a decrease of intraoperative bleeding and an increase in the number of retrieved lymph nodes and the R0 resection rate using RAMPS as compared with SRPS.

Language：English  

Language：English  

BACKGROUND:The number of operations performed by a surgeon may be an indicator of surgical skill. The hand motions made by a surgeon also reflect skill and level of expertise. We hypothesized that the hand motions of expert and novice surgeons differ significantly, regardless of whether they are familiar with specific tasks during an operation.METHODS:This study compared 11 expert surgeons, each of whom had performed &GT;100 laparoscopic procedures, and 27 young surgeons, each of whom had performed &LT;15 laparoscopic procedures. Each examinee performed a specific skill assessment task, in which instrument motion was monitored using magnetic tracking system. We analyzed the paths of the centers of gravity of the tips of the needle holders and the relative paths of the tips using two mathematical methods of detrended fluctuation analysis and unstable periodic orbit analysis.RESULTS:Detrended fluctuation analysis showed that the exponent in the function
describing the initial scaling exponent (α1) differed significantly for experts and novices, being close to 1.0 and 1.5, respectively (P &LT; 0.01). This indicated that the expert group had a greater long-range coherence with an intrinsic sequence and smooth continuity among a series of motions. Likewise, unstable periodic orbit analysis showed that the second period of unstable orbit was significantly longer for experts in comparison with novices (P &LT; 0.01). This demonstrates mathematically that the hands of experts are more stable when performing laparoscopic procedures.CONCLUSIONS:Objective evaluation of hand motion during a simulated laparoscopic procedure showed a significant difference between experts and novices.

Language：Japanese  

INTRODUCTION:We evaluated the differences in instrument manipulation skills between expert laparoscopic surgeons and novices.METHODS:Six expert surgeons who had performed more than 500 laparoscopic surgeries and one skilled instructor at Kyushu University Training Center for Minimally Invasive Surgery, and 20 medical students who had experienced no laparoscopic surgery were enrolled. A new skill assessment task was designed using zippers on an unstable, mobile platform in a box trainer. The examinees were asked to close the zippers, while trying to avoid moving the platform. The path lengths of the tips of the instruments and of the platform were measured, and the performance time was also recorded. Surgical skill score was calculated from the correlation between the path lengths of the instruments and that of the platform, in addition to the performance time.RESULTS:The path lengths of the tips of both instruments and of the platform were significant
ly shorter in the experts than in the novices (all p &LT; 0.05). The performance time was also significantly shorter for experts than novices (p &LT; 0.05). The surgical skill score was significantly higher for experts than novices (p &LT; 0.01).CONCLUSION:The differences in the instrument manipulation skills between expert laparoscopic surgeons and novices could therefore be evaluated using our surgical skill scoring system.

Language：English  

The cell surface protein Transmembrane 4 L6 family member 1 (TM4SF1) has been detected in various tumors, and its expression on tumor cells is implicated in cancer cell metastasis and patient prognosis. The role of TM4SF1 in malignant tumors remains poorly understood, particularly in pancreatic cancer. We performed immunohistochemical staining to analyze the expression of TM4SF1 in resected pancreatic tissues and investigated the correlation between TM4SF1 expression and prognosis. The function of TM4SF1 in the invasion and migration of pancreatic cancer cells was analyzed in vitro using an RNA interference technique. In pancreatic cancer tissues, TM4SF1 expression was detected in cancer cells, and patients with high tumor levels of TM4SF1 showed longer survival times than those with low TM4SF1 levels (P=0.0332). In vitro, reduced TM4SF1 expression enhanced the migration (P<0.05) and invasion (P<0.05) of pancreatic cancer cells partially via decreased E-cadherin expression. TM4SF1 protein levels were also reduced after TGF-β1-induced epithelial-mesenchymal transition (EMT).TM4SF1 expression is associated with better prognosis in pancreatic cancer. Loss of TM4SF1 contributes to the invasion and migration of pancreatic cancer cells.

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BACKGROUND:

Esophagogastric anastomosis after esophagectomy has been performed with a variety of techniques during the past decade. However, anastomotic leakage and stricture are still important clinical problems after esophagogastric anastomosis, causing burdensome symptoms and poor quality of life. Herein, we describe a novel cervical end-to-side triangulating esophagogastric anastomoasis using linear stapler.

METHODS:

A total of 90 patients (85 % male; mean age 63 years) with thoracic esophageal cancer who underwent cervical end-to-side esophagogastric triangular anastomosis using a linear stapler after minimally invasive esophagectomy between November 2006 and April 2013 were retrospectively reviewed.

RESULTS:

The median operation time was 602 min (range 424-936 min). The volume of blood loss during the entire operative procedure was 127 ml (range 0-700 ml). There were no cases of anastomotic leakage in this study, although four patients (4.4 %) developed dysphagia associated with benign anastomotic stricture formation. All patients with a benign anastomotic stricture underwent balloon dilation, which resulted in improvement in their symptoms.

CONCLUSIONS:

Considering the absence of anastomotic leakage and low rate of anastomotic stricture formation in this study, our modified triangular esophagogastric anastomosis technique appears promising and may contribute to reduced morbidity and mortality rates following esophagectomy.

DOI： 10.1007/s00268-014-2925-0

Language：English  

Abstract
BACKGROUND:
The purpose of this study was to develop a new objective assessment system for the suture ligature method employed in the laparoscopic intestinal anastomosis model. Suturing skills were evaluated objectively using this system.
METHODS:
This study compared 17 expert surgeons, each of whom had performed >500 laparoscopic procedures, with 36 novice surgeons, each of whom had performed <15 laparoscopic procedures. Each examinee performed a specific skill assessment task using an artificial model that mimics living tissue, which is linked with the Suture Simulator Instruction Evaluation Unit. The model used internal air pressure measurements and image processing to evaluate suturing skills. Five criteria were used to evaluate the skills of participants.
RESULTS:
The volumes of air pressure leak in the expert and novice groups were 21.13 ± 6.68 and 8.51 ± 8.60 kPa, respectively. The numbers of full-thickness sutures in the expert and novice groups were 2.94 ± 0.24 pairs and 2.47 ± 0.77 pairs, respectively. Suture tensions in the expert and novice groups were 60.99 ± 11.81 and 80.90 ± 16.63 %, respectively. The areas of wound-opening in the expert and novice groups were 1.76 ± 2.17 and 11.06 ± 15.37 mm(2), respectively. The performance times in the expert and novice groups were 349 ± 120 and 750 ± 269 s, respectively. Significant differences between the expert and novice groups for each criterion were observed. The acceptable range of values for each criterion except for the number of full-thickness sutures was statistically defined by the performance of the expert group.
CONCLUSIONS:
Our system is useful for the quantitative assessment of suturing skill in laparoscopic surgery. We believe that this system is a useful tool for training and assessment of laparoscopic surgeons.

Language：English  

Abstract
BACKGROUND AND STUDY AIMS:
The conventional procedure of endoscopic submucosal dissection (ESD) is technically demanding. This study investigated the efficiency of novel articulating devices (maximum diameter 2.6 mm), which can be used with commercially available, standard endoscopes.
PATIENTS AND METHODS:
In an ex vivo comparative study, eight endoscopists were divided into novices and experienced operators, and performed ESD using new devices and the conventional setup. An in vivo animal experiment was performed by two experts. Procedure times for incision and dissection were recorded, and unit times for circumferential length and area of specimens were calculated.
RESULTS:
All procedures were successfully completed with en bloc resection. In the ex vivo study, the unit procedure times for incision and dissection by novices were significantly shorter using the new system (P < 0.01 and P < 0.05), whereas there was no significant difference for experienced endoscopists. Perforation occurred during one procedure in which the new system was used. The in vivo experiments were successfully completed without adverse events.
CONCLUSIONS:
ESD using novel articulating devices was feasible. These devices were able to reduce the procedure time for novices.

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We describe the development of neuropilin 1-binding peptide (iRGD)-nanocages that specifically target human pancreatic cancer cells in which an iRGD is joined to the surface of naturally occurring heat shock protein (HSP) cages. Using a genetic engineering approach, the iRGD domain was joined to the C-terminal region of the HSP cage using flexible linker moieties. The characteristics of the interdomain linkages between the nanocage and the iRGD domain play an important role in the specificity and affinity of the iRGD-nanocages for their target cells. An engineered L30-iRGD-nanocage with 30 amino acid linkers, (GGS)10, showed greater binding affinity for pancreatic cancer cells relative to that of other linkers. Furthermore, a moderately hydrophobic anticancer drug, OSU03012, was successfully incorporated into the L30-iRGD-nanocage by heating the mixture. The OSU03012-loaded L30-iRGD-nanocage induced cell death of pancreatic cancer cells by activating
the caspase cascade more effectively than the same concentrations of free OSU03012. The iRGD-nanocages show great potential as a novel nanocarrier for pancreatic cancer-targeted drug delivery.

Language：English  

OBJECTIVE:

The aims of this study were to investigate the GNAS mutational status in pancreatic intraductal papillary mucinous neoplasm (IPMN) with and without distinct pancreatic ductal adenocarcinoma (PDAC) and to evaluate the significance of GNAS analysis using duodenal fluid (DF) in patients with IPMN.

METHODS:

The clinicopathologic features of 110 patients with IPMN including 16 with distinct PDAC were reviewed. The GNAS status in the IPMN tissue and 23 DF specimens was assessed by sensitive mutation scanning methods.

RESULTS:

The GNAS mutation rate in IPMN with distinct PDAC was significantly lower than that in IPMN without PDAC (4/16, 25%, vs 61/94, 65%; P = 0.0047). By multivariate analysis, GNAS wild-type and gastric type IPMNs were significantly associated with distinct PDAC. Of 45 GNAS wild-type IPMNs, 10 (43%) of 23 gastric type IPMNs had distinct PDAC, whereas only 2 (9%) of 22 non-gastric type IPMNs had distinct PDAC (P = 0.017). The GNAS status in DF was consistent with that in tissue in 21 (91%) of 23 patients.

CONCLUSIONS:

Distinct PDACs frequently develop in the pancreas with gastric type IPMN without GNAS mutations. Duodenal fluid DNA test would predict the GNAS status of IPMN, whereas the detection of the gastric subtype using noninvasive test remains to be determined.This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License, where it is permissible to download and share the work provided it is properly cited. The work cannot be changed in any way or used commercially. http://creativecommons.org/licenses/by-nc-nd/3.0.

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Background: Main duct intraductal papillary mucinous neoplasms (MD-IPMNs) may occur in one or multiple segments of the pancreatic duct. Unlike multifocal branch duct (BD)-IPMNs, the clonality of multisegmental MD-IPMNs remains unclear. GNAS mutations are common and specific for IPMNs, and mutational assessment might be useful to determine the clonality of IPMNs as well as to detect high-risk IPMN with distinct ductal adenocarcinoma (PDAC). Our aim was to clarify clonality using GNAS status in multisegmental MD-IPMNs.Methods: Medical records of 70 patients with MD-IPMN were retrospectively reviewed. Histological subtypes and KRAS/GNAS mutations were investigated, and the clonal relationships among multisegmental MD-IPMNs were assessed. Mutational analysis was performed using high-resolution melting analysis and subsequent Sanger/pyrosequencing.Results: Thirteen patients had multiple synchronous and/or metachronous lesions. Seven of these 13 patients had multip
le MD-IPMNs; three had multiple MD-IPMNs and distinct BD-IPMNs; one had multiple MD-IPMNs and a distinct PDAC; one had a solitary MD-IPMN, BD-IPMN, and PDAC; and one had a solitary MD-IPMN and PDAC. KRAS/GNAS mutations were consistent in 10 of 11 multisegmental MD-IPMNs, while MD-IPMNs, BD-IPMNs, and PDACs tended to show different mutational patterns. The frequency of malignant IPMNs was significantly higher in the multisegment cohort; malignant IPMNs constituted 90% (9/10) of the multiple cohort and 56% (32/57) of the solitary cohort (P=0.04). Mutant GNAS was more frequently observed in the intestinal subtype (94%) than the others.Conclusions: MD-IPMNs can be characterized by monoclonal skip progression. Close attention should be paid to the possible presence of skip areas during/after partial pancreatectomy.

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Anterior gradient 2 (AGR2), a member of the protein disulfide isomerase family, has been implicated in various cancers including pancreatic ductal adenocarcinoma (PDAC) and is known to promote cancer progression. However, the prognostic value of AGR2 expression and the interaction with epithelial-mesenchymal transition (EMT) remain unclear. We investigated the clinical significance of AGR2 and EMT markers in PDAC patients by immunohistochemical analyses. Although AGR2 expression was not observed in normal pancreas, all pancreatic precursor neoplastic lesions were positive for AGR2, even at the earliest stages, including pancreatic intraepithelial neoplasia-1A, AGR2 expression was reduced in 27.7% (54/195 cases) of PDAC patients. AGR2 downregulation correlated with EMT markers (vimentin overexpression and reduced membranous E-cadherin expression), high Union for International Cancer Control stage (P<0.0001), high histological cellular grade (P<0.0001), and adverse outcome (P<0.0001). In vitro, targeted silencing of AGR2 in cancer cells using siRNA reduced cell proliferation, colony formation, cell invasiveness, and migration, but did not alter EMT markers. To confer a more aggressive phenotype and induce EMT in PDAC cells, we co-cultured PDAC cell lines with primary-cultured pancreatic stellate cells (PSCs) and found that AGR2 was downregulated in co-cultured PDAC cells compared with PDAC monocultures. Treatment with transforming growth factor beta-1 (TGF-β), secreted from PSCs, decreased AGR2 expression, whereas inhibition of TGF-β signaling using recombinant soluble human TGF-β receptor type II and TGF-β-neutralizing antibodies restored AGR2 expression. We conclude that AGR2 downregulation is a useful prognostic marker, induced by EMT, and that secreted TGF-β from PSCs may partially contribute to AGR2 downregulation in PDAC patients. AGR2 downregulation does not induce EMT or a more aggressive phenotype, but is a secondary effect of these processes in advanced PDAC.Laboratory Investigation advance online publication, 24 November 2014; doi:10.1038/labinvest.2014.138.

Language：English  

OBJECTIVES:

To clarify the recurrence pattern after resection of main duct intraductal papillary mucinous neoplasms (MD-IPMNs) using molecular analyses and determine the most adequate treatment strategy.

BACKGROUND:

The most appropriate resection line for MD-IPMNs remains an unresolved issue.

METHODS:

Medical records of 56 patients with pancreatectomy were retrospectively reviewed. Histological subtypes and Kras/GNAS mutations were assessed in patients with recurrence in the remnant pancreas.

RESULTS:

Forty-nine patients underwent partial pancreatectomy and 7 underwent total pancreatectomy. Thirty-six patients (64%) had malignant MD-IPMNs. Recurrence was observed in 7 of 49 patients (14%), including 6 with malignant IPMNs and 1 with pancreatic ductal adenocarcinoma, all of whom underwent remnant pancreatectomy. The cumulative disease-specific survival rate of patients with pancreatic recurrence was greater than that of patients with extrapancreatic recurrence (P < 0.001). Although the pancreatic margin status at the initial operation did not affect the pancreatic recurrence rate, all 4 recurrent IPMNs examined had histological subtypes and Kras/GNAS mutations identical to those of the initial lesions. Four patients experienced recurrence in the remnant pancreas or systemic recurrence after resection of high-grade dysplasia of MD-IPMN. Three of the 56 patients had concomitant pancreatic ductal adenocarcinomas and MD-IPMNs.

CONCLUSIONS:

One-step total pancreatectomy can be avoided, and remnant total pancreatectomy would lead to favorable outcomes even in patients with pancreatic recurrence, some cases of which seem to involve residual lesions. Postoperative surveillance of high-grade dysplasia should be performed as if malignant, and close attention should be paid to the occurrence of concomitant pancreatic ductal adenocarcinomas in patients with MD-IPMNs.

Language：English  

Frozen human tissues are necessary for research purposes, but tissue banking methods have not changed for more than a decade. Many institutions use cryovial tubes or plastic molds with an optimal cutting temperature compound. However, these methods are associated with several problems, such as samples sticking to one another and the need for a larger storing space. We established an efficient tissue freezing and storing procedure ("tissue tablet method") applicable to both molecular analysis and frozen tissue microarray. Tissue samples were chopped into tiny fragments and embedded into tablet-shaped frozen optimal cutting temperature compound using our original tissue-freezing plate. These tablets can be sectioned and stored in cryovial tubes. We compared the tissue quality of tablet-shaped samples with that of conventional optimal cutting temperature blocks and found no significant difference between them. Tissue microarray is a key method to utilize tissue-banking specimens. However, most tissue microarrays require the coring out of cylindrically shaped tissues from formalin-fixed, paraffin-embedded tissue blocks. Antigenic changes and mRNA degradation are frequently observed with formalin-fixed, paraffin-embedded samples. Therefore, we have applied tablet-shaped samples to construct frozen tissue microarrays with our original mounting base. Constructed tissue microarray sections showed good morphology without obvious artifact and good immunohistochemistry and in situ hybridization results. These results suggest that the quality of arrayed samples was sufficiently appropriate for research purposes. In conclusion, the tissue tablet method and frozen tissue microarray procedure can save time, provides easy tissue handling and processing, and satisfies the demands of research methodologies and tissue banking.

Language：English  

BACKGROUND:

Since its widespread acceptance for the treatment of early gastric cancer, laparoscopic gastrectomy has been gaining popularity as a treatment option for advanced gastric cancer. However, laparoscopic total gastrectomy (LTG) with splenectomy is seldom performed, because of its difficulty of removal of station 10 lymph nodes; splenectomy is technically essential for complete removal of these lymph nodes. The purpose of this study was to describe the details of the LTG procedure and to evaluate the short- and long-term outcomes of LTG with splenectomy.

METHODS:

Of 725 consecutive patients with gastric cancer who underwent laparoscopic gastrectomy with lymph node dissection in our institution from January 1996 to December 2012, 18 consecutive patients who underwent LTG with splenectomy were enrolled in this study.

RESULTS:

No operative mortality occurred, and the pathological margins were free from cancer cells in all patients. The mean operation time was 388 min (range 324-566 min). The mean volume of blood loss was 45 ml (range 5-347 ml), and the mean number of dissected lymph nodes was 51 (range 40-105). Postoperative morbidity occurred in six patients (33.3 %) (each with grade B postoperative pancreatic fistula, postoperative bleeding, chylous ascites, atelectasis, ileus, and intra-abdominal infection). Five patients (27.8 %) developed recurrence (four in the peritoneum and one in the liver), and the overall 3- and 5-year survival rates were 83.0 and 72.6 %, respectively.

CONCLUSIONS:

Considering the 0 % mortality rate and low rates of postoperative morbidity and locoregional recurrence, LTG with splenectomy is technically and oncologically acceptable. This procedure can be expanded to include advanced gastric cancer, which generally requires splenectomy for lymph node dissection.

Language：English  

BACKGROUND:Metacognition is the knowledge about one's own methods of perceiving, remembering, thinking, and acting. This study determined the significance of metacognitive skills in laparoscopic surgery with the aim of applying the findings in a laparoscopic surgery training program.MATERIAL AND METHODS:Eighteen medical students with no experience in laparoscopic surgery (novice group) and eight expert surgeons who had each performed &GT;100 laparoscopic surgeries (expert group) were enrolled. The examinees in each group performed an evaluation task using a virtual reality simulator and answered questions about the task.RESULTS:The longest performance times, longest path lengths, and most frequent tissue damage occurred at 135° in the novice group and at 180° in the expert group. The greatest recognition of task difficulties, impatience, and irritation occurred at 135° in the novice group and at 180° in the expert group. Th
ere were statistically significant correlation coefficients between the instrument path length and task difficulty (metacognition) at 135° (R = 0.74, p = 0.03) and 180° (R = 0.79, p = 0.02) in the expert group, but there were no significant correlations in the novice group.CONCLUSION:We elucidated the significance of metacognitive skills in laparoscopic surgery. A training program should include recognition feedback systems.

Language：English  

A thickened, enhanced cyst wall on imaging examinations is one of the "worrisome features" described in the consensus guidelines for management of intraductal papillary mucinous neoplasm of the pancreas (IPMN). Podoplanin (PDPN) expression by cancer-associated fibroblasts is known to be an indicator of poor prognosis in some types of cancer. We performed immunohistochemical staining for alpha-smooth muscle actin (α-SMA) in IPMN lesions and determined the pathological wall thickness by measuring the thinnest and thickest α-SMA-positive parts of the wall of the largest cyst in each case, and the mean of these two values was recorded as the wall thickness. The thickness of the pathological wall increased with progression from IPMN with low-grade dysplasia to IPMN with an invasive carcinoma. The pathological wall was thicker in IPMN with main duct involvement, nongastric-type IPMN, and IPMN with mural nodules. We also stained for PDPN and assessed the thickness of cyst wall staining as for α-SMA. The thickness of the PDPN-positive cyst wall varied in a pattern similar to the thickness of the α-SMA-positive pathological cyst wall. PDPN-positive stromal fibroblasts in the invasive component of IPMN-IC were evaluated as a ratio to α-SMA-positive fibroblasts. A high ratio (>50 %) of PDPN-positive stromal fibroblasts was a predictor of poor outcome. PDPN expression in the cyst wall correlates with the progression of IPMN. PDPN may be a significant prognostic marker of IPMN-IC.

Language：English  

Myofibroblasts in the stroma of pancreatic cancers promote tumor proliferation, invasion and metastasis by increasing extracellular matrix and secretion of several growth factors. In contrast, the role of myofibroblasts at peritoneally disseminated sites of pancreatic cancer has not yet been determined. This study was designed to assess the role of myofibroblasts at peritoneally disseminated sites of pancreatic cancer. Three primary cultures of human peritoneal myofibroblasts (hPMFs) were established from disseminated sites of pancreatic cancer and their interactions with the SUIT-2 and CAPAN-1 human pancreatic cancer cell lines were analyzed in vitro. Using a model in BALB/c nu/nu mice, we compared the dissemination ability of intraperitoneally implanted pancreatic cancer cells, with and without hPMFs, and examined the presence of green fluorescent protein (GFP)-labeled hPMFs at peritoneally disseminated sites in mice. hPMFs significantly promoted the migration and invasion of pancreatic cancer cells (P<0.05), while the cancer cells significantly promoted the migration and invasion of hPMFs (P<0.05). In vivo, the number of peritoneally disseminated nodules, more than 3 mm in size, was significantly greater in mice implanted with cancer cells plus hPMFs compared to mice implanted with cancer cells alone, with GFP-labeled hPMFs surviving in the peritoneal cavity of the former. hPMFs promote the peritoneal dissemination of pancreatic cancer. The cancer-stromal cell interaction in the peritoneal cavity may be a new therapeutic target to prevent the dissemination of pancreatic cancer.

Language：English  

BACKGROUND:

Micro RNAs (miRNAs) are small noncoding RNAs that have gained attention as key molecules in the malignant characteristics of cancers, and several recent investigations also have identified some miRNAs as potential key regulators to inhibit the malignant characteristics of tumors. MiRNA-373 (miR-373) has recently been reported to induce E-cadherin, which is a key regulator of epithelial-mesenchymal transition (EMT). However, the role of miR-373 in the characteristics of cancer cells is not still well known.

METHODS:

We investigated the expression levels of miR-373 in pancreatic cancer cell lines and its effect on the invasiveness of pancreatic cancer by using in vitro and in vivo models. We also analyzed the expression of miR-373 using formalin-fixed paraffin-embedded (n = 152) and microdissected frozen (n = 57) samples from pancreatic tissues.

RESULTS:

The levels of miR-373 expression were low in pancreatic cancer cell lines. In formalin-fixed paraffin-embedded and microdissected frozen samples, miR-373 expression was significantly down-regulated in pancreatic cancer compared with that in healthy pancreas (P < 0.001 and P = 0.005, respectively). We also found that reexpression of miR-373 repressed transforming growth factor-β-induced EMT, leading to inhibition of invasiveness of cancer cells. Furthermore, reexpression of miR-373 significantly inhibited peritoneal dissemination in vivo (P < 0.001).

CONCLUSIONS:

MiR-373 is down-regulated in pancreatic cancer, and its reexpression represses the invasiveness of pancreatic cancer cells.

Language：English  

Abstract
OBJECTIVES:
Gemcitabine resistance (GR) is one of the critical issues for therapy for pancreatic cancer, but the mechanism still remains unclear. Our aim was to increase the understanding of GR by metabolic profiling approach.
METHODS:
To establish GR cells, 2 human pancreatic cancer cell lines, SUIT-2 and CAPAN-1, were exposed to increasing concentration of gemcitabine. Both parental and chemoresistant cells obtained by this treatment were subjected to metabolic profiling based on liquid chromatography-mass spectrometry.
RESULTS:
Multivariate statistical analyses, both principal component analysis and orthogonal partial least squares discriminant analysis, distinguished metabolic signature of responsiveness and resistance to gemcitabine in both SUIT-2 and CAPAN-1 cells. Among significantly different (P < 0.005) metabolite peaks between parental and GR cells, we identified metabolites related to several metabolic pathways such as amino acid, nucleotide, energy, cofactor, and vitamin pathways. Decreases in glutamine and proline levels as well as increases in aspartate, hydroxyproline, creatine, and creatinine levels were observed in chemoresistant cells from both cell lines.
CONCLUSIONS:
These results suggest that metabolic profiling can isolate distinct features of pancreatic cancer in the metabolome of gemcitabine-sensitive and GR cells. These findings may contribute to the biomarker discovery and an enhanced understanding of GR in pancreatic cancer.

Language：English  

BACKGROUND:

The benefits and feasibility of laparoscopic surgery for remnant gastric cancer are still unclear. The purpose of this study was to describe the detailed procedure and to evaluate the clinical short-term outcomes of laparoscopic total gastrectomy (LTG) compared with open total gastrectomy (OTG) for remnant gastric cancer (RGC).

METHODS:

Of 1,247 consecutive patients who underwent gastrectomy for gastric cancer in our department at Kyushu University Hospital from January 1996 to May 2012, 22 patients who underwent successful curative resection of RGC with precise nodal dissection were enrolled in this study. Twelve patients underwent LTG and the remaining ten patients underwent OTG. We analyzed the clinical short-term outcomes of LTG and compared the results between LTG and OTG groups to evaluate the safety and feasibility of LTG.

RESULTS:

Twelve patients with RGC successfully underwent LTG without open conversion and morbidity. The mean operation time of LTG, 362.3 ± 68.4 min, was significantly longer than that of OTG (p = 0.0176), but the mean blood loss of LTG, 65.8 ± 62 g, was smaller than that of OTG (p < 0.01). The mean postoperative times to resumption of water and food intake were significantly shorter in the LTG group than in the OTG group (p < 0.01). The overall 3-year survival rate was comparable between the LTG and OTG groups (77.8 vs. 100 %; p = 0.9406).

CONCLUSIONS:

This study shows that LTG is a feasible and reliable procedure for the treatment of RGC in terms of short-term outcomes.

Language：English  

OBJECTIVES:

Glucagon-like peptide 1 (GLP-1) interacts with its specific high-affinity receptor, glucagon-like peptide 1 receptor (GLP-1R), and induces cellular growth and inhibition of apoptosis in pancreatic β cells. The aim of this study was to investigate the significance of GLP-1R expression in pancreatic neuroendocrine tumors (PNETs).

METHODS:

Glucagon-like peptide 1 receptor expression was semiquantitatively evaluated by immunohistochemical staining in 50 resected PNETs, and the correlation between the GLP-1R expression and clinicopathologic features was investigated.

RESULTS:

There were 23 PNETs with positive expression and 27 PNETs with negative expression of GLP-1R. Positive expression of GLP-1R was more frequently observed in insulinoma than in gastrinoma and nonfunctioning tumor (P < 0.05). Although expression status of GLP-1R did not affect the prognosis of the patients with PNETs (P = 0.82), most of the metastatic sites such as lymph node and liver showed positive staining for GLP-1R (8 of 11 PNETs, 73%).

CONCLUSIONS:

Glucagon-like peptide 1 receptor would be a diagnostic marker of insulinoma and might become a molecular target for treatment of metastatic PNETs and hormonal regulation of insulin.

Language：English  

AbstractOBJECTIVES: Aberrant expression of several microRNAs (miRs) has been reported in various neoplasms including intraductal papillary mucinous neoplasms (IPMNs) of the pancreas. MicroRNA-196a (miR-196a) is up-regulated in Barrett esophagus (characterized by intestinal metaplasia) and in colorectal cancer; this relationship between intestinal characteristics and miR-196a might also be applicable to intestinal-type IPMNs. The aim of this study was to evaluate whether intestinal-type IPMNs can be discriminated from non-intestinal-type IPMNs by the expression level of miR-196a in tissue and pancreatic juice samples.METHODS: Thirty-seven formalin-fixed paraffin-embedded tissue samples (including 3 of normal pancreatic ducts) and 36 pancreatic juice samples were obtained. The expression level of miR-196a measured by quantitative reverse transcription-polymerase chain reaction assays was compared between intestinal-type and non-intestinal-type IPMNs.RESULTS: Mi
croRNA-196a expression in intestinal-type IPMN tissue samples (n = 18) was significantly higher than that of non-intestinal-type IPMNs (n = 16) (P &LT; 0.001). Similarly, miR-196a expression in pancreatic juice samples of intestinal-type IPMNs (n = 6) was significantly higher than that of non-intestinal-type IPMNs (n = 30) (P = 0.008), and the sensitivity and specificity for prediction of intestinal-type IPMNs using pancreatic juice samples were both 83%.CONCLUSIONS: Elevated expression of miR-196a in pancreatic juice samples is predictive of intestinal-type IPMNs.

Language：English  

BACKGROUND:

CD166, also known as activated leukocyte cell adhesion molecule (ALCAM), is expressed by various cells in several tissues including cancer. However, the role of CD166 in malignant tumors is controversial, especially in pancreatic cancer. This study aimed to clarify the role and significance of CD166 expression in pancreatic cancer.

METHODS:

We performed immunohistochemistry and flow cytometry to analyze the expression of CD166 in surgical pancreatic tissues and pancreatic cancer cell lines. The differences between isolated CD166+ and CD166- pancreatic cancer cells were analyzed by invasion and migration assays, and in mouse xenograft models. We also performed quantitative RT-PCR and microarray analyses to evaluate the expression levels of CD166 and related genes in cultured cells.

RESULTS:

Immunohistochemistry revealed high expression of CD166 in pancreatic cancer tissues (12.2%; 12/98) compared with that in normal pancreas controls (0%; 0/17) (p = 0.0435). Flow cytometry indicated that CD166 was expressed in 33.8-70.2% of cells in surgical pancreatic tissues and 0-99.5% of pancreatic cancer cell lines. Invasion and migration assays demonstrated that CD166- pancreatic cancer cells showed stronger invasive and migratory activities than those of CD166+ cancer cells (p<0.05). On the other hand, CD166+ Panc-1 cells showed a significantly stronger colony formation activity than that of CD166- Panc-1 cells (p<0.05). In vivo analysis revealed that CD166+ cells elicited significantly greater tumor growth than that of CD166- cells (p<0.05) in both subcutaneous and orthotopic mouse tumor models. mRNA expression of the epithelial-mesenchymal transition activator Zeb1 was over-expressed in CD166- cells (p<0.001). Microarray analysis showed that TSPAN8 and BST2 were over-expressed in CD166+ cells, while BMP7 and Col6A1 were over-expressed in CD166- cells.

CONCLUSIONS:

CD166+ pancreatic cancer cells are strongly tumorigenic, while CD166- pancreatic cancer cells exhibit comparatively stronger invasive and migratory activities. These findings suggest that CD166 expression is related to different functions in pancreatic cancer cells.

Language：English  

OBJECTIVES:

The aim of this study was to establish a minimally invasive and simple screening test for detection of pancreatic ductal adenocarcinoma (PDAC) using duodenal juice (DJ).

METHODS:

Duodenal juice was collected prospectively before endoscopic retrograde cholangiopancreatography in 46 patients. A protease inhibitor was not added to the samples collected during the initial 2.5 minutes but was added in the latter 2.5 minutes. Thereafter, secretin was administered intravenously, and DJ was subsequently collected for additional 10 minutes. The sensitivities of carcinoembryonic antigen (CEA), S100 calcium-binding protein P (S100P), and interleukin 8 in DJ and pancreatic juice were assessed.

RESULTS:

There were 30 patients with PDAC and 16 with benign lesions. It was possible to collect an adequate amount of DJ without secretin administration. In the PDAC group, CEA concentrations in DJ were significantly higher than those in the benign group, even without the use of a protease inhibitor. S100P levels in DJ in the PDAC group were significantly higher than those in the benign group in the presence of the protease inhibitor.

CONCLUSIONS:

Duodenal juice collection during routine upper endoscopy and assessments of CEA and S100P in DJ might become a useful screening test for detection of PDAC.

Language：English  

Abstract

BACKGROUND:

Pancreatic cancer (PC), a hypovascular tumor, thrives under hypoxic conditions. Pancreatic stellate cells (PSCs) promote PC progression by secreting soluble factors, but their functions in hypoxia are poorly understood. This study aimed to clarify the effects of hypoxic conditions on the interaction between PC cells and PSCs.

METHODS:

We isolated human PSCs from fresh pancreatic ductal adenocarcinomas and analyzed functional differences in PSCs between normoxia (21% O(2)) and hypoxia (1% O(2)), including expression of various factors related to tumor-stromal interactions. We particularly analyzed effects on PC invasiveness of an overexpressed molecule-connective tissue growth factor (CTGF)-in PSCs under hypoxic conditions, using RNA interference techniques.

RESULTS:

Conditioned media from hypoxic PSCs enhanced PC cell invasiveness more intensely than that from normoxic PSCs (P < 0.01). When co-cultured with PSCs, PC cell invasion was more enhanced under hypoxia than under normoxia (P < 0.05). Among various soluble factors, which were related to invasiveness, CTGF was one of the overexpressed molecules in hypoxic PSCs. A higher level of CTGF expression was also found in supernatant of hypoxic PSCs than in supernatant of normoxic PSCs. PC cell invasiveness was reduced by CTGF knockdown in hypoxic PSCs co-cultured with PC cells (P < 0.05).

CONCLUSION:

Hypoxia induces PSCs' secretion of CTGF, leading to enhancement of PC invasiveness. CTGF derived from hypoxia-stimulated PSCs may be a new therapeutic target for pancreatic cancer.

Language：English  

Abstract


PURPOSE:

The role of gastrectomy for patients with positive peritoneal cytology, but a negative macroscopic peritoneal implant (P-/cy+), remains unclear. The aim of this study was to evaluate laparoscopic gastrectomy for P-/cy+ patients.

METHODS:

This study reviewed a prospectively maintained gastric cancer database of gastric-cancer patients those underwent surgical resection. P-/cy+ gastric cancer that had invaded the subserosa, or deeper layers, of the stomach wall without distant organ metastases was considered operable in this institution. P-/cy+ patients underwent either open or laparoscopic gastrectomy with D2 lymphadenectomy. The short-term results were examined to assess differences in outcome between the two groups.

RESULTS:

Eighteen P-/cy+ patients without distant organ metastases underwent surgery between 2000 and 2010. Laparoscopic gastrectomy was performed in nine patients and open gastrectomy in nine patients. The estimated blood loss was significantly smaller, the resumption of food intake earlier, and the length of postoperative hospital stay shorter in the patients that underwent laparoscopic gastrectomy than in the patients that underwent open gastrectomy. There were no significant differences in the 2-year survival rates between the groups.

CONCLUSION:

Laparoscopic gastrectomy for P-/cy+ patients is a minimally invasive and safe oncologic procedure with good short-term results.

Language：English  

BACKGROUND:

Although laparoscopic distal gastrectomy has been widely accepted in clinical practice, laparoscopic total gastrectomy (LTG) is not yet familiar because of the difficulty in esophagojejunostomy. The purpose of this study was to evaluate perioperative and short-term outcomes of our procedure of intracorporeal gastrojejunostomy using linear staplers after LTG.

METHODS:

Of 98 consecutive patients who underwent LTG for gastric cancer in our department between August 2002 and December 2010, 94 patients underwent esophagojejunostomy with a linear stapling device. After October 2007, we modified the esophagojejunostomy; ie, the most recent 57 patients underwent transection of the esophagus in the ventrodorsal direction and insertion of a linear stapler from the anterior wall of the Roux limb to the posterior wall so as to make an inverted T-shaped anastomosis. We evaluated the results in these 57 patients (recent group) and compared them with the results in the earlier 37 patients (early group).

RESULTS:

The mean operative time in the recent group was 368 to 94.6 min, and the mean estimated blood loss was 57 to 33 g; both were comparable with those in the early group. Neither open conversion nor intraoperative complications were encountered. Two patients experienced anastomotic leakage in the earlier group, but anastomotic leakage did not occur in the recent group. No mortality was encountered.

CONCLUSION:

We herein report our procedure of intracorporeal gastrojejunostomy using linear staplers after LTG. Our procedure of esophagojejunostomy using linear staplers is safe and feasible and has acceptable morbidity.

Copyright © 2013 Mosby, Inc. All rights reserved.

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This study describes the applications of a naturally occurring small heat shock protein (Hsp) that forms a cage-like structure to act as a drug carrier. Mutant Hsp cages (HspG41C) were expressed in Escherichia coli by substituting glycine 41 located inside the cage with a cysteine residue to allow conjugation with a fluorophore or a drug. The HspG41C cages were taken up by various cancer cell lines, mainly through clathrin-mediated endocytosis. The cages were detected in acidic organelles (endosomes/lysosomes) for at least 48 hours, but none were detected in the mitochondria or nuclei. To generate HspG41C cages carrying doxorubicin (DOX), an anticancer agent, the HspG41C cages and DOX were conjugated using acid-labile hydrazone linkers. The release of DOX from HspG41C cages was accelerated at pH 5.0, but was negligible at pH 7.2. The cytotoxic effects of HspG41C-DOX against Suit-2 and HepG2 cells were slightly weaker than those of free DOX, but the ef
fects were almost identical in Huh-7 cells. Considering the relatively low release of DOX from HspG41C-DOX, HspG41C-DOX exhibited comparable activity towards HepG2 and Suit-2 cells and slightly stronger cytotoxicity towards Huh-7 cells than free DOX. Hsp cages offer good biocompatibility, are easy to prepare, and are easy to modify; these properties facilitate their use as nanoplatforms in drug delivery systems and in other biomedical applications.

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PURPOSE:In pediatric endoscopic surgery, a limited view and lack of tactile sensation restrict the surgeon's abilities. Moreover, in pediatric oncology, it is sometimes difficult to detect and resect tumors due to the adhesion and degeneration of tumors treated with multimodality therapies. We developed an augmented reality (AR) navigation system based on preoperative CT and MRI imaging for use in endoscopic surgery for pediatric tumors.METHODS:The patients preoperatively underwent either CT or MRI with body surface markers. We used an optical tracking system to register the reconstructed 3D images obtained from the CT and MRI data and body surface markers during surgery. AR visualization was superimposed with the 3D images projected onto captured live images. Six patients underwent surgery using this system.RESULTS:The median age of the patients was 3.5 years. Two of the six patients underwent laparoscopic surgery, two patients underwent thoracoscopi
c surgery, and two patients underwent laparotomy using this system. The indications for surgery were local recurrence of a Wilms tumor in one case, metastasis of rhabdomyosarcoma in one case, undifferentiated sarcoma in one case, bronchogenic cysts in two cases, and hepatoblastoma in one case. The average tumor size was 22.0±14.2 mm. Four patients were treated with chemotherapy, three patients were treated with radiotherapy before surgery, and four patients underwent reoperation. All six tumors were detected using the AR navigation system and successfully resected without any complications.CONCLUSIONS:The AR navigation system is very useful for detecting the tumor location during pediatric surgery, especially for endoscopic surgery.

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OBJECTIVE:: To identify a high-risk group of patients with pancreatic ductal adenocarcinoma (PDAC), independently arising in the pancreas with intraductal papillary mucinous neoplasm (IPMN), using histopathologic subtypes. BACKGROUND:: Pathologic features of IPMN with distinct PDAC, including histopathologic subtypes of IPMN and PDAC phenotypes, have not been well characterized. Mucin expression patterns and the mutational status of GNAS and KRAS are useful to explore the relationship between these 2 lesion types. METHODS:: Clinicopathologic data of 179 resected IPMNs and 180 resected PDACs without IPMNs as a control group were reviewed. IPMNs were classified into 4 grades (low-grade, intermediate-grade, high-grade dysplasia, and an associated invasive carcinoma) and 4 subtypes (gastric, intestinal, pancreatobiliary, and oncocytic). The expression of MUC1, MUC2, MUC5AC, MUC6, and CDX2 was investigated by immunohistochemistry in IPMNs and PDACs with an
d without IPMNs. The mutational status of GNAS and KRAS was evaluated by cycle sequencing in PDACs and pre-/coexisting IPMNs. RESULTS:: Twenty-six synchronous or metachronous PDACs were identified in 20 patients (11.2%) with IPMNs. Occurrence of concomitant PDACs was more frequently observed in gastric-type IPMNs (18/110, 16.4%) compared with intestinal (1/49, 2.0%), pancreatobiliary (1/17, 5.9%), or oncocytic-type (0/3, 0%) (P = 0.047). Both PDACs with and without IPMNs were frequently positive for MUC1, MUC5AC, and MUC6 expression, as assessed by immunohistochemistry, but were negative for MUC2 and CDX2. The mucin-staining patterns were similar to those of invasive tubular adenocarcinoma arising from gastric-type IPMNs. Mutation of GNAS within codon 201 was not detected in PDACs and gastric-type IPMNs, whereas most of these exhibited KRAS mutations. However, the R201H GNAS mutation was detected in 1 intestinal-type IPMN with distinct PDAC. CONCLUSIONS:: Mucin expression pa
tterns demonstrate that PDAC without GNAS mutations of an aggressive phenotype frequently arise in the pancreas with benign gastric-type IPMN in the absence of GNAS mutations.

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BACKGROUND:The purpose of this research was to investigate the usefulness of three-dimensional (3D) endoscopy compared with two-dimensional (2D) endoscopy in neuroendoscopic surgeries in a comparative study and to test the clinical applications.METHODS:Forty-three examinees were divided into three groups according to their endoscopic experience: novice, beginner, or expert. Examinees performed three separate tasks using 3D and 2D endoscopy. A recently developed 3D high-definition (HD) neuroendoscope, 4.7 mm in diameter (Shinko Optical Co., Ltd., Tokyo, Japan) was used. In one of the three tasks, we developed a full-sized skull model of acrylic-based plastic using a 3D printer and a patient's thin slice computed tomography data, and evaluated the execution time and total path length of the tip of the pointer using an optical tracking system. Sixteen patients underwent endoscopic transnasal transsphenoidal pituitary surgery using both 3D and 2D endoscop
y.RESULTS:Horizontal motion was evaluated using task 1, and anteroposterior motion was evaluated with task 3. Execution time and total path length in task 3 using the 3D system in both novice and beginner groups were significantly shorter than with the 2D system (p &LT; 0.05), although no significant difference between 2D and 3D systems in task 1 was seen. In both the novice and beginner groups, the 3D system was better for depth perception than horizontal motion. No difference was seen in the expert group in this regard. The 3D HD endoscope was used for the pituitary surgery and was found very useful to identify the spatial relationship of carotid arteries and bony structures.CONCLUSIONS:The use of a 3D neuroendoscope improved depth perception and task performance. Our results suggest that 3D endoscopes could shorten the learning curve of young neurosurgeons and play an important role in both general surgery and neurosurgery.

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Improving poor outcomes in patients with pancreatic cancer requires a greater understanding of the biological mechanisms contributing to radioresistance. We, therefore, sought to identify genes involved in the radioresistance of pancreatic cancer cells. Two pancreatic cancer cell lines, CFPAC-1 and Capan-1, were repeatedly exposed to radiation, establishing two radioresistant cell lines. Gene expression profiling using cDNA microarrays was performed to identify genes responsible for radioresistance. The levels of expression of mRNAs encoded by selected genes and their correlation with radiation dose resulting in 50% survival rate were analyzed in pancreatic cancer cell lines. The radiation dose resulting in a 50% survival rate was significantly higher in irradiated (IR) compared to parental CFPAC-1 cells (8.31 ± 0.85 Gy vs. 2.14 ± 0.04 Gy, P<0.0001), but was lower in IR compared with parental Capan-1 cells (2.66 ± 0.24 Gy vs. 2.25 ± 0.03 Gy, P=0.04). cDNA microarray analysis identified 4 genes, including S100 calcium binding protein A4 (S100A4), overexpressed and 23 genes underexpressed in the IR compared with the parental cell lines. The levels of S100A4 mRNA expression were correlated with radiation dose resulting in a 50% survival rate (Pearson's test, R2=0.81, P=0.0025). S100A4 mRNA expression may predict radioresistance of pancreatic cancer cells and may play an important role in the poor response of pancreatic cancer cells to radiation therapy.

Language：English  

BACKGROUND:

Interactions between cancer cells and surrounding cancer-associated fibroblasts (CAFs) play an important role in cancer progression. Invasive ductal carcinoma (IDC) of the pancreas is characterized by abundant fibrous connective tissue called desmoplasia. Podoplanin (PDPN) is a lymphatic vessel marker (D2-40), and expression of PDPN by stromal CAFs has been reported to be a prognostic indicator in various types of cancer.

METHODS:

Expression of PDPN in pancreatic IDCs was assessed by immunohistochemical examination in 105 patients who underwent pancreatic resection. Primary CAFs were established from pancreatic cancer tissue obtained by surgery. Quantitative reverse transcription-polymerase chain reaction and flow cytometric analysis were performed to investigate PDPN expression in CAFs. We sorted CAFs according to PDPN expression, and analyzed the functional differences between PDPN+ CAFs and PDPN- CAFs using indirect co-culture with pancreatic cancer cell lines. We also investigated the culture conditions to regulate PDPN expression in CAFs.

RESULTS:

PDPN expression in stromal fibroblasts was associated with lymphatic vessel invasion (P = 0.0461), vascular invasion (P = 0.0101), tumor size ≥ 3 cm (P = 0.0038), histological grade (P = 0.0344), Union for International Cancer Control classification T stage (P = 0.029), and shorter survival time (P < 0.0001). Primary CAFs showed heterogeneous PDPN expression in vitro. Moreover, migration and invasion of pancreatic cancer cell lines (PANC-1 and SUIT-2) were associated with PDPN expression in CAFs (P < 0.01) and expression of CD10, matrix metalloproteinase (MMP) 2, and MMP3. In cultured CAFs, PDPN positivity changed over time under several conditions including co-culture with cancer cells, different culture media, and addition of growth factor.

CONCLUSIONS:

PDPN-expressing CAFs enhance the progression of pancreatic IDC, and a high ratio of PDPN-expressing CAFs is an independent predictor of poor outcome. Understanding the regulation of the tumor microenvironment is an important step towards developing new therapeutic strategies.

Language：English  

Pancreatic stellate cells (PSCs), which are implicated in desmoplasia in pancreatic cancer, enhance the malignancy of cancer cells and confer resistance to established treatments. We investigated whether the antifibrotic agent pirfenidone can suppress desmoplasia and exert anti-tumor effects against pancreatic cancer. Primary PSCs were established from pancreatic cancer tissue obtained during surgery. In vitro, pirfenidone inhibited the proliferation, invasiveness, and migration of PSCs in a dose-dependent manner. Although supernatants of untreated PSCs increased the proliferation, invasiveness, and migration of pancreatic cancer cells (PCCs), supernatants of pirfenidone-treated PSCs decreased these effects. Exposure to PCC supernatant increased the production of platelet-derived growth factor-A, hepatic growth factor, collagen type I, fibronectin, and periostin in PSCs, which was significantly reduced by pirfenidone. Mice were subcutaneously implante
d with PCCs (SUIT-2 cells) andPSCs into the right flank and PCCs alone into the left flank. Oral administration of pirfenidone to these mice significantly reduced tumor growth of co-implanted PCCs and PSCs, but not of PCCs alone. Pirfenidone also decreased the proliferation of PSCs and the deposition of collagen type I and periostin in tumors. In mice with orthotopic tumors consisting of PCCs co-implanted with PSCs, pirfenidone suppressed tumor growth, reduced the number of peritoneal disseminated nodules, and reduced the incidence of liver metastasis. Pirfenidone in combination with gemcitabine more effectively suppressed orthotopic tumor growth compared with pirfenidone or gemcitabine alone. In conclusion, our findings indicate that pirfenidone is a promising antitumor agent for pancreatic cancer, owing to its suppression of desmoplasia through regulating PSCs.

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OBJECTIVES: CD105 expression correlates with prognosis for several cancers. However, its significance in pancreatic cancer is unclear.METHODS: We analyzed CD105 expression in resected pancreatic cancer tissue and pancreatic cancer cell lines, compared the properties of CD105 and CD105 cells using quantitative RT-PCR and migration assays, and evaluated the relationship between CD105 cells and pancreatic stellate cells (PSCs).RESULTS: Immunohistochemistry showed that the frequency of CD105 expression was higher in pancreatic cancer than that in normal tissue (8% vs 0%, respectively). In flow cytometry, CD105 was expressed in pancreatic cancer cells, whereas weak CD105 expression was detected in normal pancreatic ductal epithelial cells. Quantitative RT-PCR showed that E-cadherin mRNA expression was suppressed and vimentin mRNA was overexpressed in CD105 cells (P &LT; 0.05). Migration of CD105 cancer cells was strongly enhanced (more than that of CD105 c
ells) in coculture with PSCs (P &LT; 0.05). CD105 expression did not correlate to clinicopathologic characteristics or the Kaplan-Meier survival analysis.CONCLUSIONS: Suppression of an epithelial marker and overexpression of a mesenchymal marker suggest that epithelial-mesenchymal transition is induced in CD105 pancreatic cancer cells. CD105 pancreatic cancer cell migration is strongly enhanced by PSCs, suggesting that these cells play a role in the pancreatic cancer microenvironment.

Language：English  

BACKGROUND:

Pancreatic cancer is associated with a devastating prognosis, partially because of its aggressive metastatic ability. Identification of prognostic markers of metastasis would be useful in the clinical management of postoperative patients with pancreatic cancer. Mal, T-cell differentiation protein 2 (MAL2) has been identified as a molecule predictive of metastases; the clinical relevance of MAL2 in pancreatic cancer is unknown.

METHODS:

Orthotopic human pancreatic cancer xenografts from the pancreatic cancer cell line SUIT-2 were established in nude mice. Only liver metastasis was harvested and cultured. These metastatic cycles were repeated 5 times to establish a highly metastatic cell line, termed metastatic SUIT-2 (MS). We investigated proliferation and motility of MS cells compared with those of the parent SUIT-2. Microarray analysis was performed to investigate differences in gene expression. We also performed immunohistochemical analysis of 89 formalin-fixed, paraffin-embedded human pancreatic cancer tissue samples to investigate the clinical significance of MAL2 expression.

RESULTS:

MS cells showed a greater metastatic rate after orthotopic implantation than parental SUIT-2. MS cells had increased motility but decreased proliferation compared with parental SUIT-2. Microarray analyses showed that 26 genes were significantly upregulated (>10-fold) in MS cells compared with parental SUIT-2, particularly MAL2 expression. Immunohistochemical analysis showed that high expression of MAL2 was associated with a lesser survival of postoperative patients (P = .03) and a high rate of distant metastasis (P = .008).

CONCLUSION:

We characterized a newly established pancreatic cancer cell line with highly metastatic potential. MAL2 is a promising predictive marker for distant metastasis and short survival in patients with resected pancreatic cancer.

Copyright © 2013 Mosby, Inc. All rights reserved.

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Abstract
Kindlin-1 is a novel focal adhesion protein that belongs to the kindlin family. Expression of kindlin-1 has recently been reported in lung and colon cancers, but there have been no studies on its expression in pancreatic cancer. This study aimed to investigate the expression and function of kindlin-1 in pancreatic cancer. Quantitative RT-PCR of Kindlin-1 mRNA was performed in various pancreatic cancer cell lines as well as normal pancreatic epithelial cells and fibroblasts. Immunohistochemical analysis of kindlin-1 was performed for pancreatic cancer tissues. The effects of kindlin-1 on the proliferation, migration and invasion of pancreatic cancer cells were investigated using an RNA interference technique. Kindlin-1 mRNA was highly expressed in the pancreatic cancer cell lines, but only slightly expressed in normal pancreatic epithelial cells and fibroblasts. The Kindlin-1 protein was heterogeneously expressed in the cytoplasm and membrane of pancreatic cancer cells, while normal ductal epithelial cells and stromal cells showed no expression. In vitro experiments involving knockdown of kindlin-1 in AsPC-1 and KP-2 cells revealed that the migratory and invasive abilities of the cells were significantly decreased (P<0.001), while the proliferation abilities were not affected. The present findings suggest that kindlin-1 expression is involved in the progression of pancreatic cancer via enhancement of cell migration and invasion.

Language：English  

PURPOSE:In endoscopic surgery, limited views and lack of tactile sensation restrict the surgeon's abilities and cause stress to the surgeon. Therefore, an intra-operative navigation system is strongly recommended. We developed an augmented reality (AR) navigation system based on preoperative CT imaging. The purpose of this study is to evaluate the usefulness, feasibility, and accuracy of this system using laparoscopic splenectomy in children.METHODS:Volume images were reconstructed by three-dimensional (3D) viewer application. We used an optical tracking system for registration between volume image and body surface markers. The AR visualization was superimposed preoperative 3D CT images onto captured laparoscopic live images. This system was applied to six cases of laparoscopic splenectomy in children. To evaluate registration accuracy, distances from the marker position to the volume data were calculated.RESULTS:The operator recognized the hidden vas
cular variation of the splenic artery and vein, accessory spleen, and pancreatic tail by overlaying an image onto a laparoscopic live image. The registration accuracy of six cases was 5.30 ± 0.08, 5.71 ± 1.70, 10.1 ± 0.60, 18.8 ± 3.56, 4.06 ± 1.71, and 7.05 ± 4.71.CONCLUSION:This navigation system provides real-time anatomical information, which cannot be otherwise visualized without navigation. The registration accuracy was acceptable in clinical operation.

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BACKGROUND/AIMS:

Endoscopic submucosal dissection (ESD) is gaining wider acceptance for the treatment of early gastric carcinoma (EGC) and its indication has been extended to mucosal gastric carcinoma with undifferentiated component in some institutes. Our aims were to confirm the frequency of lymph node metastasis in such cases and clarify the demarcation in indications for ESD.

METHODOLOGY:

We evaluated medical data of 287 patients with mucosal gastric carcinoma who underwent surgical resection between 1996 and 2008. The tumours were histologically classified into purely differentiated (PD), differentiated-predominant mixed (DPM), undifferentiated-predominant mixed (UPM) and purely undifferentiated (PU) types.

RESULTS:

Lymph node metastasis was identified in seven (2.4%) of the 287 patients and was detected more frequently in UPM (10%, two of 20) and PU (4%, four of 98), compared with PD (none of 148) (p=0.01 and 0.02, respectively). In mixed-type carcinoma, size was a significant risk factor for lymph node metastasis (p=0.04).

CONCLUSIONS:

It might be better to select gastrectomy rather than ESD for the treatment of mucosal gastric carcinoma with an undifferentiated component.

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Abstract
In this study, we examined the cytotoxic effects of combination therapy with zoledronic acid (ZOL) and gemcitabine (GEM) on pancreatic cancer cells in vitro and in vivo. Four human pancreatic cancer cell lines were treated with ZOL, GEM or a combination of both, and the effects of the respective drug regimens on cell proliferation, invasion and matrix metalloproteinase (MMP) expression were examined. A pancreatic cancer cell line was also intrasplenically or orthotopically implanted into athymic mice and the effects of these drugs on tumor metastasis and growth in vivo were evaluated by histologic and immunohistochemical analyses. Combination treatment with low doses of ZOL and GEM efficiently inhibited the proliferation (P < 0.001) and invasion (P < 0.001) of pancreatic cancer cells in vitro. Western blotting assay revealed that MMP-2 and MMP-9 expression levels were decreased after ZOL treatment. In vivo, combined treatment significantly inhibited tumor growth (P < 0.05) and the development of liver metastasis (P < 0.05). These data revealed that, when used in combination, ZOL and GEM have significant antitumor, anti-metastatic and anti-angiogenic effects on pancreatic cancer cells. In the present study, we first reported the significance of the combination treatment of ZOL and GEM in pancreatic cancer using an in vivo model. These data show promise for the future application of this drug regimen in patients with pancreatic cancer.

© 2011 Japanese Cancer Association.

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Abstract


BACKGROUND:

Extracellular matrix (ECM) remodeling is predominantly mediated by fibroblasts using intracellular and extracellular pathways. Although it is well known that extracellular degradation of the ECM by proteases derived from cancer cells facilitates cellular invasion, the intracellular degradation of ECM components by cancer cells has not been clarified. The aim of this study was to characterize collagen internalization, which is the initial step of the intracellular degradation pathway in pancreatic cancer cells, in light of epithelial-mesenchymal transition (EMT).

METHODOLOGY/PRINCIPAL FINDINGS:

We analyzed the function of collagen internalization in two pancreatic cancer cell lines, SUIT-2 and KP-2, and pancreatic stellate cells (PSCs) using Oregon Green 488-gelatin. PSCs had a strong ability for collagen uptake, and the pancreatic cancer cells also internalized collagen although less efficiently. The collagen internalization abilities of SUIT-2 and KP-2 cells were promoted by EMT induced by human recombinant transforming growth factor β1 (P<0.05). Expression of Endo180, a collagen uptake receptor, was high in mesenchymal pancreatic cancer cell lines, as determined by EMT marker expression (P<0.01). Quantitative RT-PCR and western blot analyses showed that Endo180 expression was also increased by EMT induction in SUIT-2 and KP-2 cells. Endo180 knockdown by RNA interference attenuated the collagen uptake (P<0.01) and invasive abilities (P<0.05) of SUIT-2 and KP-2 cells.

CONCLUSIONS/SIGNIFICANCE:

Pancreatic cancer cells are capable of collagen internalization, which is enhanced by EMT. This ECM clearance system may be a novel mechanism for cellular invasion and a potential therapeutic target in pancreatic cancer.

Language：English  

Abstract
BACKGROUND:
There is increasing evidence that microRNAs are differentially expressed in many types of cancers. Despite progress in analyses of microRNAs in several types of cancers, the functional contributions of microRNAs to pancreatic cancer remain unclear.
METHODS:
In the present study, the expression levels of specific microRNAs identified by microarray analyses were examined in a panel of 15 pancreatic cancer cell lines. We then investigated the functional roles of these microRNAs in the proliferation and invasion of pancreatic cancer cells.
RESULTS:
Based on the microarray data, we found frequent and marked overexpression of miR-10a, miR-92, and miR-17-5p in pancreatic cancer cell lines. Microdissection analyses revealed that miR-10a was overexpressed in pancreatic cancer cells isolated from a subset of primary tumors (12 of 20, 60%) compared with precursor lesions and normal ducts (P<.01). In vitro experiments revealed that miR-10a inhibitors decreased the invasiveness of pancreatic cancer cells (P<.01), but had no effect on their proliferation. Inhibition of HOXA1, a target of miR-10a, promoted the invasiveness of pancreatic cancer cells (P<.01).
CONCLUSIONS:
The present data suggest that miR-10a is overexpressed in a subset of pancreatic cancers and is involved in the invasive potential of pancreatic cancer cells partially via suppression of HOXA1.

Language：English  

Protein nanocages are self-organized complexes of oligomers whose three-dimensional architecture can been determined in detail. These structures possess nanoscale inner cavities into which a variety of molecules, including therapeutic or diagnostic agents, can be encapsulated. These properties yield these particles suitable for a new class of drug delivery carrier, or as a bioimaging reagent that might respond to biochemical signals in many different cellular processes. We report here the design, synthesis, and biological characterization of a hepatocyte-specific nanocage carrying small heat-shock protein. These nanoscale protein cages, with a targeting peptide composed of a preS1 derivative from the hepatitis B virus on their surfaces, were prepared by genetic engineering techniques. PreS1-carrying nanocages showed lower cytotoxicity and significantly higher specificity for human hepatocyte cell lines than other cell lines in vitro. These results sug
gested that small heat-shock protein-based nanocages present great potential for the development of effective targeted delivery of various agents to specific cells.

Language：English  

BACKGROUND:

A dilated orifice of the duodenal papilla found during screening endoscopy or ERCP is well-known as one of the specific findings of intraductal papillary mucinous neoplasm (IPMN). However, its clinical significance is still unclear.

OBJECTIVE:

To assess the diagnostic significance of a dilated orifice of the duodenal papilla and evaluate whether this could be a factor predictive of malignancy or a subtype of IPMN.

DESIGN:

Retrospective study.

SETTING:

University hospital.

PATIENTS:

This study involved 149 patients who underwent pancreatectomy for IPMN between January 1987 and June 2011.

INTERVENTION:

ERCP.

MAIN OUTCOME MEASUREMENTS:

The rate of malignant and intestinal type IPMNs in patients with and without papillary dilation.

RESULTS:

A dilated orifice of the duodenal papilla was significantly associated with intestinal type IPMN (P < .001), but this finding could not predict the malignant grade of IPMN (P = .13). Multivariate analysis revealed that a dilated orifice was a significant factor for predicting intestinal type in both main duct (P = .01) and branch duct IPMNs (P < .001).

LIMITATIONS:

The validity of the definition of papillary dilation, selection bias, and a retrospective study.

CONCLUSION:

A dilated orifice of the duodenal papilla could be a significant factor for predicting intestinal type IPMN. This may lead to better clinical management of patients with IPMN.

Copyright © 2012 American Society for Gastrointestinal Endoscopy. Published by Mosby, Inc. All rights reserved.


Comment in
Pancreatic cancer: Dilated orifice of the duodenal papilla predicts intestinal type IPMN. [Nat Rev Gastroenterol Hepatol. 2012]

Language：English  

Abstract
Pancreatic stellate cells (PSCs) play a crucial role in the aggressive behavior of pancreatic cancer. Although heterogeneity of PSCs has been identified, the functional differences remain unclear. We characterized CD271⁺ PSCs in human pancreatic cancer. Immunohistochemistry for CD271 was performed for 31 normal pancreatic tissues and 105 pancreatic ductal adenocarcinomas (PDACs). We performed flow cytometry and quantitative RT-PCR, and assessed CD271 expression in PSCs isolated from pancreatic tissues and the changes in CD271 expression in PSCs cocultured with cancer cells. We also investigated the pattern of CD271 expression in a SCID mouse xenograft model. In the immunohistochemical analyses, the CD271-high staining rates in pancreatic stroma in normal pancreatic tissues and PDACs were 2/31 (6.5%) and 29/105 (27.6%), respectively (p = 0.0069). In PDACs, CD271⁺ stromal cells were frequently observed on the edge rather than the center of the tumors. Stromal CD271 high expression was associated with a good prognosis (p = 0.0040). Flow cytometric analyses demonstrated CD271-positive rates in PSCs were 0-2.1%. Quantitative RT-PCR analyses revealed that CD271 mRNA expression was increased in PSCs after coculture with pancreatic cancer cells. However, the level of CD271 mRNA expression subsequently decreased after the transient increase. Furthermore, CD271 mRNA expression was decreased in PSCs migrating toward pancreatic cancer cells through Matrigel. In the xenograft model, CD271⁺ PSCs were present at tumor margins/periphery and were absent in the tumor core. In conclusion, CD271 was expressed in PSCs around pancreatic tumors, but not in the center of the tumors, and expression decreased after long coculture with pancreatic cancer cells or after movement toward pancreatic cancer cells. These findings suggest that CD271⁺ PSCs appear at the early stage of pancreatic carcinogenesis and that CD271 expression is significantly correlated with a better prognosis in patients with PDAC.

DOI： 0.1371/journal.pone.0052682

Language：English  

BACKGROUND: Although gemcitabine is the standard treatment for pancreatic cancer, this particular type of cancer develops rapidly and has intrinsic chemoresistance. Chemoresistance plays a critical role in tumor progression, invasion and migration. Nevertheless, the effect of adenoviral therapy on chemoresistant cancer cells has not been studied. In this study, we compared the efficacy of adenoviral therapy in parental and chemoresistant pancreatic cancer cells.

MATERIALS AND METHODS: To establish gemcitabine-resistant cells, pancreatic cancer SUIT2 cells were exposed to increasing concentrations of gemcitabine. Both parental and chemoresistant cells were infected with adenoviruses expressing either green fluorescent protein (Ad-GFP) or the hepatocyte growth factor antagonist, NK4 (Ad-NK4). To investigate the transduction efficacy, GFP expression and NK4 concentrations were measured and an invasion assay was used to investigate the efficacy of the adenoviral therapy.

RESULTS: The 50% inhibitory concentration of gemcitabine was <10 nM in the parental SUIT-2 cells, while it was >1 μM in gemcitabine-resistant cells. A large number of gemcitabine-resistant cells were GFP-positive compared with only a small number of parental cells (p<0.05). The NK4 expression level was significantly higher in gemcitabine-resistant cells than in parental cells (p<0.05). The supernatant from Ad-NK4-infected gemcitabine-resistant cells significantly inhibited the invasion of cancer cells compared with that from Ad-NK4-infected parental cells (p<0.05).

CONCLUSION: Both the efficiency of transduction and the therapeutic efficacy of adenoviral therapy were higher in gemcitabine-resistant cells than in parental cells, suggesting that adenoviral gene therapy is more effective in patients with gemcitabine-resistant pancreatic cancer.

Language：Japanese  

Abstract：鏡視下手術における2次元映像の奥行き情報の欠如は,技術精度が要求される外科医にとって大きなストレスである.今回,われわれはドーム型立体内視鏡映像(3DD)提示システムを臨床症例に導入し評価したので報告する.2010年1月までに15例に使用し,9名の熟練した術者にアンケート調査を行った.3DDは奥行き情報を提示することで縫合結紮など煩雑な操作を容易にし,また実質臓器の病変などの凹凸の認識にも有効であった.また,屈曲可能鉗子の使用や単孔式手術など,奥行き情報が重要な場面で特に有用であった.高度な技術をもつ外科医がより難度の高い手術を行う際の手術支援システムとして3DDは今後普及していくと考えられる.(著者抄録)

Language：English  

Many preclinical studies have shown the potential of adenovirus-based cancer gene therapy. However, successful translation of these promising results into the clinic has not yet been achieved. Pancreatic ductal adenocarcinoma (PDAC) is characterized by abundant desmoplastic stroma, and tumor-stromal cell interactions play a critical role in tumor progression. Therefore, we hypothesized that tumor-stroma interactions reduce the efficacy of adenoviral therapy. We investigated the effect of fibroblasts on adenovirus-based gene therapy using SUIT-2 and PANC-1 pancreatic cancer cells cultured with or without fibroblast-conditioned culture supernatant then infected with Ad-LacZ. After 48 h, the cells were stained for β-galactosidase. The results showed that the number of β-galactosidase-positive cells was significantly reduced after culture with fibroblast-conditioned supernatant (P < 0.05). Because the hepatocyte growth factor (HGF)/MET pathway plays an important role in tumor-stroma interactions we next investigated the involvement of this pathway in tumor-stroma interactions leading to the decreased efficacy of adenoviral therapy. SUIT-2 cells were cultured with or without SU11274 (a MET inhibitor) and/or fibroblast-conditioned culture supernatant, then infected with Ad-GFP. After 48 h, GFP-positive cells were counted. The number of GFP-positive cells in cultures containing fibroblast-conditioned supernatant plus SU11274 was significantly greater than in cultures without SU11274. In conclusion, our results suggest that stromal cells in PDAC reduce the efficacy of adenoviral therapy through a mechanism involving the HGF/MET pathway. Control of such tumor-stroma interactions may lead to improvements in adenoviral gene therapy for PDAC.

Language：English  

Abstract
CD24 is a molecule involved in cell adhesion and tumor metastasis. The aims of this study were (1) to evaluate the association between CD24 expression and the progression of intraductal papillary mucinous neoplasms of the pancreas and (2) to investigate the association between CD24 expression in pancreatic cancer and the prognosis of patients who underwent curative pancreatectomy. Immunohistochemical analysis of CD24 was performed for 95 intraductal papillary mucinous neoplasms of the pancreas and 83 pancreatic cancers. We investigated the association between CD24 expression and the histologic grade of intraductal papillary mucinous neoplasms of the pancreas, the clinicopathologic parameters of pancreatic cancers, and the survival time of pancreatic cancer patients who underwent pancreatectomy. The positive rates of CD24 expression in intraductal papillary mucinous adenoma, borderline intraductal papillary mucinous neoplasm, noninvasive intraductal papillary mucinous carcinoma, and invasive intraductal papillary mucinous carcinoma were 5 (20%) of 24, 12 (48%) of 25, 10 (43%) of 23, and 15 (65%) of 23, respectively. The CD24-positive rates were significantly higher in borderline intraductal papillary mucinous neoplasm and intraductal papillary mucinous carcinoma compared with intraductal papillary mucinous adenoma (P = .046 and P = .007, respectively). The staining scores, which were determined from the percentage of stained cells and the staining intensity, were significantly higher in invasive intraductal papillary mucinous carcinoma than in noninvasive intraductal papillary mucinous carcinoma (P = .043). In the pancreatic cancers, higher tumor stage (P = .007), nodal metastasis (P = .021), and higher-grade tumors (P < .001) were more frequent in the CD24-positive group compared with the CD24-negative group. CD24 expression was associated with shorter survival in univariate analysis (P = .028) However, based on the multivariate analysis, the CD24 expression was not associated with survival. In conclusion, CD24 is involved in the progression of intraductal papillary mucinous neoplasms of the pancreas and in the malignant behavior of pancreatic cancers.

Language：English  

Abstract
Although CD133 has been reported to be a promising colon cancer stem cell marker, the biological functions of CD133+ colon cancer cells remain controversial. In the present study, we investigated the biological differences between CD133+ and CD133- colon cancer cells, with a particular focus on their interactions with cancer-associated fibroblasts, especially CD10+ fibroblasts. We used 19 primary colon cancer tissues, 30 primary cultures of fibroblasts derived from colon cancer tissues and 6 colon cancer cell lines. We isolated CD133+ and CD133- subpopulations from the colon cancer tissues and cultured cells. In vitro analyses revealed that the two populations showed similar biological behaviors in their proliferation and chemosensitivity. In vivo analyses revealed that CD133+ cells showed significantly greater tumor growth than CD133- cells (P=0.007). Moreover, in cocultures with primary fibroblasts derived from colon cancer tissues, CD133+ cells exhibited significantly more invasive behaviors than CD133- cells (P<0.001), especially in cocultures with CD10+ fibroblasts (P<0.0001). Further in vivo analyses revealed that CD10+ fibroblasts enhanced the tumor growth of CD133+ cells significantly more than CD10- fibroblasts (P<0.05). These data demonstrate that the in vitro invasive properties and in vivo tumor growth of CD133+ colon cancer cells are enhanced in the presence of specific cancer-associated fibroblasts, CD10+ fibroblasts, suggesting that the interactions between these specific cell populations have important roles in cancer progression. Therefore, these specific interactions may be promising targets for new colon cancer therapies.

Language：English  

Abstract
Gemcitabine (GEM) is the standard treatment for advanced/metastatic pancreatic cancer. However, there is a substantial subset of patients in whom the efficacy of GEM, when used as a single agent, is inadequate. Recently, the 5-fluorouracil (5-FU) prodrugs capecitabine and S-1 have been used as an alternative, either alone or in combination with GEM. The aim of the present study was to investigate the expression pattern of genes that render pancreatic cancer cells sensitive to GEM and 5-FU, and to identify markers for individualized chemotherapy, even in patients who have developed resistance. We investigated the correlation between the expression of genes associated with the metabolism of GEM and 5-FU, and sensitivity to these drugs in 15 human pancreatic cancer cell lines. We also established GEM- and 5-FU-resistant pancreatic cancer cell lines to investigate changes in the expression levels of these genes and the effects of one drug on cells resistant to the other. We found no correlation between pancreatic cancer cell sensitivity to either GEM- or 5-FU. GEM-resistant cells did not become resistant to 5-FU and vice versa. High expression of RRM1 (P=0.048) and TS x DPD (P=0.035) correlated significantly with sensitivity to GEM and 5-FU, respectively. 5-FU-resistant cells expressed significantly higher levels of TP than parental cells (P<0.05). In conclusion, pancreatic cancer cells showed no cross-resistance to GEM and 5-FU. Quantitative analyses of RRM1, TP, DPD and TS mRNA levels in pancreatic cancer cells may be useful for predicting their sensitivity to GEM and 5-FU.

Language：English  

BACKGROUND:

MicroRNAs (miRNAs) have been gaining attention as new, key molecules that contribute to carcinogenesis. In pancreatic cancer, previous profiling analyses of miRNA expression have shown that several miRNAs are differently expressed in normal and cancerous tissues. Several pancreatic cancer-specific miRNAs differed, however, in each analysis.

METHODS:

We investigated the miRNA expression profiles of the pancreatic cancer cell lines CAPAN-1 and CFPAC1 and an immortalized human normal pancreatic ductal epithelial cell line (HPDE) using a high-throughput, TaqMan, qRT-PCR array analysis. We also analyzed the expression levels of this miRNA in microdissected (n = 15) and formalin-fixed, paraffin-embedded (FFPE) (n = 115) samples from pancreatic cancers by quantitative RT-PCR. Finally, we investigated the effects of this miRNA on the invasiveness of pancreatic cancer cells.

RESULTS:

Based on the microarray analysis, miR-372, miR-146a, miR-204, miR-10a, and miR-10b showed particularly large differences (>10-fold changes) between both pancreatic cell lines and HPDE cells. Thirteen of the 15 pancreatic cancer cell lines showed 2.1- to 36.4-fold (median, 15.3-fold) greater levels of miR-10b than HPDE cells. Microdissection analysis revealed that miR-10b exhibited greater expression levels in pancreatic cancer cells (n = 5) than in normal pancreatic ductal cells (n = 10) (P < .020). Analysis of FFPE samples showed that high miR-10b expression was associated with a lesser overall survival (P = .014). Furthermore, miR-10b correlated with the invasiveness of pancreatic cancer cells (P < .01).

CONCLUSION:

miR-10b is overexpressed in pancreatic cancer and may be involved in the invasiveness in pancreatic cancer cells, thereby leading to a poor prognosis.

Language：English  

Abstract
BACKGROUND: To improve the prognosis of patients after resection of pancreatic cancer, the most appropriate and efficient treatment should be provided to specific subsets of patients. Our aim was to identify promising microRNAs as markers to predict responses to gemcitabine in patients with resected pancreatic cancer.

METHODS: Two gemcitabine-resistant pancreatic cancer cell lines were established, and global microRNA expression analyses was performed by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Eleven miRNAs were selected as putative predictive markers and analyzed by means of macrodissected formalin-fixed, paraffin-embedded samples obtained from 90 patients with or without gemcitabine treatment after resection of pancreatic cancer.

RESULTS: We identified 24 microRNAs whose expression was altered in gemcitabine-resistant cells. qRT-PCR analyses showed that patients with high miR-142-5p and miR-204 expression had significantly longer survival times than those with low miR-142-5p (P = 0.0077) and miR-204 (P = 0.0054) expression in the gemcitabine-treated group. This was not seen in the nontreated group. Multivariate analyses showed that miR-142-5p expression was an independent prognostic marker only in patients treated with gemcitabine (P = 0.034).

CONCLUSIONS: miR-142-5p is a promising predictive marker for gemcitabine response in patients with resected pancreatic cancer.

Language：English  

Abstract
OBJECTIVES: Although intestinal-type intraductal papillary mucinous carcinoma (IPMC) is reported to have a better prognosis, few studies have addressed its invasive pattern. The meaning of "minimal invasion" (MI) in IPMC also remains unclear. We investigated the prognosis of intraductal papillary mucinous neoplasm (IPMN) focusing on MI and subtypes.

METHODS: We evaluated 71 patients with IPMC among a total of 179 patients with resected IPMN.

RESULTS: Although 2 of 10 MI-IPMC patients had lymph node metastasis, there were no disease-specific deaths among the MI-IPMC patients. Minimally invasive IPMCs were more frequently observed in intestinal-type IPMC (23/33 cases) than in non-intestinal-type IPMCs (16/38 cases; P = 0.019). Among 32 patients with massively invasive IPMC, the prognosis was significantly better for patients with intestinal-type IPMC than for patients with non-intestinal-type IPMC (P = 0.013). When confined to massively invasive IPMC, tubular invasion (P < 0.001) and lymphatic (P = 0.001) or serosal (P = 0.021) invasion were less frequently observed in intestinal-type IPMC than in non-intestinal-type IPMC.

CONCLUSIONS: Invasive carcinoma derived from intestinal-type IPMN is associated with MI, colloid carcinoma, and less invasive behavior.

Language：English  

Abstract
A hypoxic microenvironment is a characteristic feature of pancreatic cancer, and induces the expressions of various genes involved in malignant behaviors. Insulin-induced gene 2 (Insig2) has recently been shown to be correlated with cellular invasion in colon cancer. However, there have been no reports regarding its expression in pancreatic cancer. In this study, we evaluated Insig2 mRNA expression and the biological function of Insig2 in pancreatic cancer. We measured Insig2 mRNA expression in cultured pancreatic cancer cell lines and invasive ductal carcinoma (IDC) cells, normal pancreatic epithelial cells, and pancreatic intraepithelial neoplasia cells obtained by laser-capture microdissection. We also investigated the effects of Insig2-targeting siRNAs on the cell proliferation and cell invasion of pancreatic cancer cell lines. All pancreatic cancer cell lines expressed Insig2 mRNA. The PANC-1 and MIA PaCa-2 pancreatic cancer cell lines showed >2-fold higher Insig2 mRNA expression levels under hypoxic conditions (1% O2) than under normoxic conditions (21% O2 ). Cell proliferation was significantly decreased in SUIT-2 cells and cell invasion was significantly decreased in SUIT-2, Capan-2, and CFPAC-1 cells after transfection of the Insig2-targeting siRNAs. In analyses of microdissected cells, cells from IDC tissues expressed significantly higher levels of Insig2 mRNA than normal pancreatic cells (P < 0.001) and pancreatic intraepithelial neoplasia cells (P = 0.082). In analyses of IDC cells, the levels of Insig2 mRNA expression were significantly higher in late-stage patients than in early-stage patients. The present data suggest that Insig2 is associated with the malignant potential of pancreatic cancer under hypoxic conditions.

Language：English  

Abstract
BACKGROUND: Advanced gastric cancers often metastasize to distant organs and the peritoneum, leading to a poor prognosis. Both invasiveness and resistance to anchorage-independent cell death (anoikis) are important factors in the process of metastasis. p600 (600-kDa protein), recently identified from a cervical cancer cell line, plays a role in both anoikis resistance and cell migration. In this study, we examined whether p600 is involved in the progression of gastric cancer.

METHODS: We used both normal gastric mucosal cells and cancer cells laser-microdissected from 42 gastric cancers and their normal counterparts, and compared their p600 mRNA expression levels with quantitative reverse transcriptase-polymerase chain reaction. We inhibited p600 expression in two gastric cancer cell lines with siRNA and examined its effect on the invasiveness and anoikis resistance both in vitro and in vivo.

RESULTS: Expression of p600 mRNA was significantly higher in gastric cancer cells than in normal mucosal cells (P = 0.027). The invasion assay revealed that invasiveness was significantly reduced by inhibition of p600 (P < 0.01). In vitro experiments revealed that cell viability and colony-formation capacity under anchorage-independent conditions were significantly reduced by inhibition of p600 (P < 0.05). In vivo experiments also showed that the establishment of intraperitoneal disseminated tumors was significantly suppressed by transient inhibition of p600 (P < 0.001).

CONCLUSIONS: Our results strongly suggest that p600 is involved in gastric cancer progression, and has a potential to be a new molecular target for gastric cancer therapy.

Language：English  

Pancreatic ductal adenocarcinoma (PDAC) still presents a major therapeutic challenge and a phase III clinical trial has revealed that the combination of gemcitabine and a human epidermal growth factor receptor type I (HER1/EGFR) targeting agent presented a significant benefit compared to treatment with gemcitabine alone. The aim of this study was to investigate EGFR mRNA expression in resected PDAC tissues and its correlation with patient prognosis. We obtained formalin-fixed paraffin-embedded (FFPE) tissue samples from 88 patients with PDAC who underwent pancreatectomy, and measured EGFR mRNA levels by quantitative real-time reverse transcription-polymerase chain reaction. The high-level EGFR group had significantly shorter disease-free-survival (p=0.029) and overall-survival (p=0.014) as shown by univariate analyses, although these did not reach statistical significance, as shown by multivariate analyses. However, we found that high EGFR expression was an independent prognostic factor in patients receiving gemcitabine-based adjuvant chemotherapy (p=0.023). Furthermore, we measured EGFR mRNA levels in 20 endoscopic ultrasound-guided fine needle aspiration (EUS-FNA) cytological specimens. Altered EGFR levels were distinguishable in microdissected neoplastic cells from EUS-FNA cytological specimens compared to those in whole cell pellets. In conclusion, quantitative analysis of EGFR mRNA expression using FFPE tissue samples and microdissected neoplastic cells from EUS-FNA cytological specimens could be useful in predicting prognosis and sensitivity to gemcitabine in PDAC patients.

Language：English  

Claudin-4, encoding a protein for tight junction formation and function, is highly overexpressed in pancreatic ductal adenocarcinoma and is also associated with invasive adenocarcinomas arising in intraductal papillary mucinous neoplasms of the pancreas. However, the expression pattern of claudin-4 during neoplastic progression of intraductal papillary mucinous neoplasms remains unknown. Using quantitative real-time reverse transcription-PCR, we analyzed claudin-4 mRNA in a panel of 14 pancreatic cancer cell lines and in formalin-fixed paraffin-embedded tissues from 80 patients with intraductal papillary mucinous neoplasms of different histological grades and papillary subtypes. Increased expression of claudin-4 was confirmed in all the pancreatic cancer cell lines tested as compared with normal ductal epithelial cells and fibroblast cultures. The claudin-4 expression was significantly higher in high-grade intraductal papillary mucinous neoplasms (borderline neoplasm and carcinoma) than in low-grade intraductal papillary mucinous neoplasms (adenoma) (P<0.0001). In addition, claudin-4 mRNA levels were significantly higher in intestinal-type intraductal papillary mucinous neoplasms than in non-intestinal-type intraductal papillary mucinous neoplasms based on papillary subclassification (P<0.0001). Our findings suggest that claudin-4 expression is associated with neoplastic progression of intraductal papillary mucinous neoplasms and, especially, with a distinct pathway to intestinal differentiation.

Language：English  

Abstract
BACKGROUND: Identification of prognostic markers would be useful in the clinical management of patients with pancreatic ductal adenocarcinoma (PDAC). The clinical relevance of claudin-4 (CLDN4), recently identified as overexpressed in PDAC, is unknown.

METHODS: Using quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR), we analyzed CLDN4 mRNA expression in a panel of 9 pancreatic cancer cell lines and formalin-fixed paraffin-embedded (FFPE) tissues from 100 patients with PDAC. The CLDN4 expression levels were then correlated with clinicopathological variables and patient outcome. We also performed immunohistochemical analysis in 20 FFPE samples of PDAC to investigate the expression of CLDN4 protein.

RESULTS: Increased expression of CLDN4 was confirmed in all the pancreatic cancer cell lines tested compared with normal ductal epithelial cells and fibroblasts. We found that low expression of CLDN4 was significantly associated with shorter survival in patients with PDAC (hazard ratio; 1.362, 95% confidence interval; 1.011-1.873, P = 0.0419). Patients with high CLDN4 expression survived longer for a median of 63.0 months, compared with 14.7 months in patients with low CLDN4 expression (P = 0.0067). In immunohistochemical analysis, the level of CLDN4 mRNA expression was significantly correlated with the expression of CLDN4 protein (P = 0.0168).

CONCLUSION: Increased expression of CLDN4 mRNA predicts better prognosis in PDAC.

Language：English  

Abstract
BACKGROUND & AIMS: Pancreatic stellate cells (PSCs) promote the progression of pancreatic cancer by producing extracellular matrix and soluble factors. However, the functional heterogeneity of PSCs has not been identified until now. Detailed characterization of the PSCs in human pancreatic cancer would provide a set of potential targets for stroma-directed therapy.

METHODS: We isolated PSCs from fresh pancreatic ductal adenocarcinoma tissue and sorted them by flow cytometry according to cell surface expression of CD10, which is a stromal prognostic marker for various tumors. We analyzed the functional differences between CD10(+) PSCs and CD10(-) PSCs.

RESULTS: Immunohistochemical analysis showed that the frequency of CD10 expression by PSCs was markedly higher in tumor tissue than in normal tissue (33.7% vs 0%, respectively, P = .028). In pancreatic ductal adenocarcinoma, CD10 expression by PSCs was associated with positive nodal metastases (P = .011) and a shorter survival time (P < .001). In vitro coculture experiments showed that CD10(+) PSCs promoted the invasiveness of pancreatic cancer cell lines, SUIT-2 and Panc-1 cells more intensively than CD10(-) PSCs. CD10(+) PSCs significantly increased the tumor growth and invasiveness of SUIT-2 cells in a murine cotransplantation model. CD10(+) PSCs secreted higher levels of matrix metalloproteinase 3 than CD10(-) PSCs, and knockdown of matrix metalloproteinase 3 in cocultured PSCs reduced the invasion of SUIT-2 and Panc-1 cells.

CONCLUSIONS: CD10(+) PSCs enhance the progression of pancreatic cancer cells. CD10(+) PSCs may be a candidate for selective therapeutic targeting in the treatment of pancreatic cancer.

Copyright © 2010 AGA Institute. Published by Elsevier Inc. All rights reserved.

Language：English  

OBJECTIVES:: Pancreatic ductal adenocarcinoma (PDAC) is often characterized by a prominent desmoplastic stroma that is induced partially by alpha-smooth muscle actin (SMA)-expressing activated pancreatic stellate cells (PSCs). This study aimed to investigate the significance of alpha-SMA expression in PDAC and the correlation between alpha-SMA mRNA levels and the patient prognosis. METHODS:: We obtained formalin-fixed, paraffin-embedded tissue samples from 109 patients with PDAC, who underwent pancreatectomy at our institution from 1992 to 2007. We measured alpha-SMA mRNA levels by quantitative real-time reverse transcription-polymerase chain reaction and investigated the association of alpha-SMA mRNA expression with clinicopathologic parameters and survival time. We also assessed the influence of activated PSCs on malignant behaviors of pancreatic cancer cells using in vitro experiments. RESULTS:: alpha-SMA immunoreactivity was detected exclusively in the stroma of PDAC. The group with high alpha-SMA expression showed a significantly shorter survival, as shown by univariate analysis (P = 0.005) and multivariate analysis (P < 0.0001). alpha-SMA-expressing activated PSCs enhanced the invasiveness, proliferation, and colony formation of pancreatic cancer cells. CONCLUSIONS:: Quantitative analysis of alpha-SMA mRNA expression using formalin-fixed, paraffin-embedded tissue samples was useful to predict the prognosis of patients with PDAC. Activated PSCs may regulate the malignant behavior of pancreatic cancer cells.

Language：English  

BACKGROUND: Secreted protein acidic and rich in cysteine (SPARC) is a protein which governs diverse cellular functions and matrix remodeling systems. It was reported that SPARC expression of peritumoral fibroblasts was correlated with poor prognosis in resectable pancreatic cancer patients in immunohistochemical analysis. However, the significance of SPARC mRNA expression in pancreatic adenocarcinoma remains unclear. MATERIALS AND METHODS: SPARC mRNA expression was evaluated in 104 formalin-fixed paraffin-embedded tissue samples of pancreatic adenocarcinoma patients with macro-dissection technique. RESULTS: High SPARC mRNA expression was associated with a poorer prognosis than low SPARC. In univariate Cox's proportion hazard model, depth of tumor invasion, lymph node metastasis, histopathological tumor grade, lymphatic invasion, vascular invasion, surgical margin, no chemotherapy and high SPARC expression were significant prognostic factors. In multivariate analysis, histopathological tumor grade, surgical margin, no chemotherapy and high SPARC expression were independent significant prognostic factors. CONCLUSION: SPARC mRNA expression is a prognostic marker for pancreatic adenocarcinoma patients.

Language：English  

Education and training to maintain medical safety are very important within clinical settings. We have established a training center for endoscopic surgery and we regularly hold a unique training course, which focuses on the development of fundamental skills. One hundred and ninety-four surgeons who participated in our training course were divided into four groups according to their experience in performing laparoscopic procedures. Group 1: 0-19 laparoscopic procedures (n=44). Group 2: 20-49 laparoscopic procedures (n=53). Group 3: 50-99 laparoscopic procedures (n=46). Group 4: more than 100 laparoscopic procedures (n=55). All subjects underwent evaluation for

Language：English  

Backgrounds: Intraductal papillary mucinous neoplasms of the pancreas (IPMNs) are subclassified based on morphological features and different immunohistochemical profiles have been identified in association with the subtypes. We previously reported that S100P was an early developmental marker of pancreatic carcinogenesis, and that there was higher S100P expression in IPMNs than in normal pancreatic ductal epithelium. However, there have been no reports on novel diagnostic markers to distinguish IPMN from non-neoplastic lesions. Materials and Methods: Surgical specimens of IPMN obtained from 105 patients were investigated using immunohistochemistry. Results: S100P expression was not detected in normal pancreatic ductal epithelium, but was detected in all IPMN cells (100%) with diffuse nuclear or nuclear/cytoplasmic staining. MUC5AC was also expressed in most of the IPMNs (102 of 105, 97%). S100P was clearly detected in minimally invasive area
s!
of IPMNs. Furthermore, S100P was diffusely expressed in the invasive component of IPMNs (32 of 32, 100%), including perineural and lymphatic invasion. On the other hand, MUC5AC was expressed in only 23 cases of 32 invasive components (P &LT; 0.01). Conclusions: These data suggest that the S100P antibody may be a useful marker for detecting all types of IPMNs including invasive lesions.

Language：English  

BACKGROUND: Recently, the microRNA-200 family was reported to affect cancer biology by regulating epithelial to mesenchymal transition (EMT). Especially, the expression of miR-200c has been shown to be associated with upregulating the expression of E-cadherin, a gene known to be involved in pancreatic cancer behavior. However, the significance of miR-200c in pancreatic cancer is unknown. METHODS: In the present study, we investigated the relationship between E-cadherin and miR-200c expression in a panel of 14 pancreatic cancer cell lines and in macro-dissected formalin-fixed paraffin-embedded (FFPE) tissue samples obtained from 99 patients who underwent pancreatectomy for pancreatic cancer. We also investigated the effects of miR-200c on the proliferation and invasion of pancreatic cancer cells. RESULTS: We found that patients with high levels of miR-200c expression had significantly better survival rates than those with low levels of miR-200c expression. We also found a remarkably strong correlation between the levels of miR-200c and E-cadherin expression. CONCLUSIONS: These data indicate that miR-200c may play a role in the pancreatic cancer biology and may be a novel marker for the prognosis of pancreatic cancer.

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BACKGROUND: Detection of aberrant microRNA (miR) expression may contribute to diagnosis and prognosis of various cancers. The aim of this study is to evaluate the correlation between miR-203 expression and prognosis of patients with pancreatic adenocarcinoma after curative resection.

METHODS: A total of 113 formalin-fixed paraffin-embedded tissue samples of pancreatic adenocarcinoma, 20 samples of chronic pancreatitis, and 8 samples of normal pancreas were obtained. We investigated the association of miR-203 expression measured by quantitative reverse-transcription polymerase chain reaction assays with clinicopathological parameters and survival times.

RESULTS: miR-203 was overexpressed in pancreatic adenocarcinoma samples compared with chronic pancreatitis (P < 0.001) and normal pancreas (P = 0.001) samples. An association between miR-203 expression and clinicopathological factors of pancreatic adenocarcinoma was not observed. On univariate analysis, the high-miR-203 group and the subgroup (20%) of cases with the highest miR-203 overexpression had significantly shorter survival time (P = 0.048 and P = 0.024, respectively). Multivariate analysis revealed that miR-203 expression was an independent predictor of poor prognosis in cases with no residual tumor (relative risk 2.298, P = 0.027).

CONCLUSIONS: miR-203 expression is a new prognostic marker in pancreatic adenocarcinoma patients.

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The microRNA-17-92 cluster is an oncogene in human B cell lymphomas and lung cancers. Previous microRNA microarray data revealed that miR-17-5p, a member of the miR-17-92 cluster, is upregulated in pancreatic cancer. However, the involvement of miR-17-5p expression in pancreatic carcinogenesis has not well been studied. In the present study, we measured the miR-17-5p expression levels in pancreatic cancer cell lines, primary cultures of normal human pancreatic ductal cells, formalin-fixed paraffin-embedded (FFPE) tissue samples derived from 80 patients who underwent pancreatectomy for pancreatic cancer and microdissected cells (including normal ductal epithelial, pancreatic intraepithelial neoplasia-1B and invasive ductal carcinoma cells) by qRT-PCR. Furthermore, we investigated the effects of upregulation of miR-17-5p expression on the proliferation and invasion of pancreatic cancer cells. We found that pancreatic cancer cells expressed higher levels of miR-17-5p than primary cultured normal ductal cells. miR-17-5p was also overexpressed in pancreatic cancer in FFPE and microdissected samples. Furthermore, analysis of macrodissected FFPE samples revealed that high miR-17-5p expression was associated with a poor prognosis (p = 0.03). In addition, in vitro experiments revealed that SUIT-2 and KP-2 pancreatic cancer cells transfected with the miR-17-5p precursor showed significantly higher cell growth ratios than the corresponding control cells (p < 0.001 and p = 0.012, respectively), as well as significantly higher numbers of invading cells (p < 0.0001 for both). The present findings suggest that miR-17-5p plays important roles in pancreatic carcinogenesis and cancer progression, and is associated with a poor prognosis in pancreatic cancer.

Language：English  

Abstract
CONTEXT: Cytological assessment of pancreatic juice is commonly used to diagnose pancreatic ductal adenocarcinoma; however, the sensitivity of cytological assessment has been reported to be low. MicroRNAs are small RNAs regulating various cellular processes and have recently been identified as possible markers of malignant diseases including pancreatic ductal adenocarcinoma.

OBJECTIVE: The purposes of this study were to prove the existence of microRNAs in pancreatic juice and to determine whether specific microRNAs in pancreatic juice could be used for detecting pancreatic ductal adenocarcinoma.

METHODS: Relative expression levels of microRNA-21 and microRNA-155 in formalin-fixed paraffin-embedded tissues of resected specimens (no. 13) and pancreatic juice samples collected using preoperative endoscopic retrograde cholangiopancreatography (no. 21) were quantified and their expression levels were then compared to pancreatic ductal adenocarcinoma and chronic pancreatitis.

RESULTS: Relative expression levels of microRNA-21 in tissue and pancreatic juice samples were significantly higher in pancreatic ductal adenocarcinoma than those in chronic pancreatitis (P=0.009 and P=0.021, respectively). The same results were obtained in the expression levels of microRNA-155 in tissue and pancreatic juice between pancreatic ductal adenocarcinoma and chronic pancreatitis (P=0.014 and P=0.021, respectively). Expression levels of microRNA-21 and microRNA-155 did not correlate with the preoperative cytological results of pancreatic juice.

CONCLUSION: MicroRNA-21 and microRNA-155 in pancreatic juice have the potential of becoming biomarkers for diagnosing pancreatic ductal adenocarcinoma.

Language：English  

BACKGROUND: Intraductal papillary mucinous neoplasm (IPMN) of the pancreas is divided into 4 subtypes: an intestinal type, a gastric type, a pancreatobiliary type, and an oncocytic type. The purposes of this study were to clarify the outcomes and the characteristics of invasive carcinoma derived from IPMN (invasive IPMC) by focusing on these subtypes with a comparison to conventional invasive ductal carcinoma (IDC) of the pancreas. METHODS: A total of 30 patients with invasive IPMC were reviewed, and the tumors were divided into 2 pathologic subtypes, intestinal and nonintestinal type. The prognosis and characteristics of the 2 subtypes were evaluated. Furthermore, the prognosis of 119 patients with conventional IDC was compared with that of patients with invasive carcinoma derived from the intestinal or nonintestinal type IPMN. RESULTS: The 5-year survival rate of patients with the nonintestinal type (0.0%) was as poor as that of patients with conventional IDC (19.9%; P = .67). The patients with the intestinal type (66.7%) had a more favorable prognosis than patients with conventional IDC (P < .001). The nonintestinal type was characterized by positive lymphatic invasion and tubular invasive pattern. CONCLUSION: Invasive carcinoma derived from the nonintestinal type IPMN characterized by lymphatic invasion and tubular invasive pattern is associated with a poor prognosis. Copyright 2010 Mosby, Inc. All rights reserved.

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Adenovirus-mediated gene therapy shows promise for cancer therapy, but transgene expression of replication-defective adenovirus may be low and transient in clinical settings. Recent reports have shown that the use of a conditionally replication-competent adenovirus (CRAd) enhanced the gene transduction of a replication-defective adenovirus vector. The control of tumor-stromal interactions has also been determined to be important in cancer therapy. In this study, we investigated the effect of the human telomerase reverse transcriptase (hTERT)-CRAd, Ad5/3hTERTE1, which possesses the tumor-specific hTERT promoter with the chimeric fiber 5/3, on the transgene expression and therapeutic efficacy of a replication-defective adenovirus vector expressing NK4 under the control of the CMV promoter, Ad-NK4. In addition, we established a new strategy to target both cancer cells and cancer-stromal interactions. Human pancreatic cancer cells were infected with Ad-NK4 and either Ad5/3hTERTE1 (CRAd-combination group) or Ad5/3hTERTLuc (control-combination group). In the CRAd-combination group, Ad-NK4-delivered transgene expression was increased, leading to an enhanced inhibitory effect on the invasion of cancer cells. In in vivo experiments, NK4 expression within tumors and its inhibitory effect on tumor growth, angiogenesis, and metastasis were enhanced in the CRAd-combination group. These results suggest that hTERT-CRAd enhances the transgene expression and therapeutic efficacies of Ad-NK4, possibly through the in-trans replication of Ad-NK4 induced by adenovirus E1 derived from co-infected hTERT-CRAd. This approach may be a promising combination therapy against advanced pancreatic cancer.

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Abstract
BACKGROUND AND AIMS: The standard palliative chemotherapy for pancreatic ductal adenocarcinoma (PDAC) is gemcitabine-based chemotherapy; however, PDAC still presents a major therapeutic challenge. The aims of this study were to investigate the expression pattern of genes involved in gemcitabine sensitivity in resected PDAC tissues and to determine correlations of gene expression with treatment outcome.

MATERIALS AND METHODS: We obtained formalin-fixed paraffin-embedded (FFPE) tissue samples from 70 patients with PDAC. Of the 70 patients, 40 received gemcitabine-based adjuvant chemotherapy (AC). We measured hENT1, dCK, CDA, RRM1, and RRM2 messenger RNA (mRNA) levels by quantitative real-time reverse transcription-polymerase chain reaction and determined the combined score (GEM score), based on the expression levels of hENT1, dCK, RRM1, and RRM2, to investigate the association with survival time. By determining the expression levels of these genes in endoscopic ultrasound-guided fine needle aspiration (EUS-FNA) cytologic specimens, we investigated the feasibility of individualized chemotherapy.

RESULTS: High dCK (P = .0067), low RRM2 (P = .003), and high GEM score (P = .0003) groups had a significantly longer disease-free survival in the gemcitabine-treated group. A low GEM score (<2) was an independent predictive marker for poor outcome to gemcitabine-based AC as shown by multivariate analysis (P = .0081). Altered expression levels of these genes were distinguishable in microdissected neoplastic cells from EUS-FNA cytologic specimens.

CONCLUSIONS: Quantitative analyses of hENT1, dCK, RRM1, and RRM2 mRNA levels using FFPE tissue samples and microdissected neoplastic cells from EUS-FNA cytologic specimens may be useful in predicting the gemcitabine sensitivity of patients with PDAC.

PMID: 20927319 [PubMed - in process]PMCID: PMC2950330Free PMC Article



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Figure 1
Quantitative analyses of mRNA associated with cellular uptake and metabolism of gemcitabine in gemcitabine-resistant cells. Quantitative analyses of hENT1 (A), dCK (B), RRM1 (C), RRM2 (D), and CDA (E) mRNA in gemcitabine-resistant cell lines (SUIT-2-GR and Capan-1-GR) and parental cells. SUIT-2-GR cells expressed sign...
Gene Expression Levels as Predictive Markers of Outcome in Pancreatic Cancer after Gemcitabine-Based Adjuvant Chemotherapy12
Neoplasia. 2010 October;12(10):807-817.Figure 2
Correlation between gemcitabine-based AC and survival time. The patients who received gemcitabine-based AC (GEMgroup) showed a significantly prolongedOStime compared with the non-AC group (P = .0017). *P < .05.
Gene Expression Levels as Predictive Markers of Outcome in Pancreatic Cancer after Gemcitabine-Based Adjuvant Chemotherapy12
Neoplasia. 2010 October;12(10):807-817.Figure 3
Correlation between the expression of each mRNA and DFS. Low dCK (P= .0067) and high RRM2 (P=.003) levels, normalized to β-actin, were associated with a shorter DFS in the GEM group (A, C, E, G). In contrast, there was no significant correlation between these gene expression levels and DFS in the non-AC group (B, D...
Gene Expression Levels as Predictive Markers of Outcome in Pancreatic Cancer after Gemcitabine-Based Adjuvant Chemotherapy12
Neoplasia. 2010 October;12(10):807-817.Figure 4
Correlation between the expression of each mRNA and OS. Low hENT1 (P = .011), low dCK (P = .0095), high RRM1 (P = .041), and high RRM2 (P = .030) levels, normalized to β-actin, were associated with a shorterOSin theGEMgroup (A, C, E, G). In contrast, there was no significant correlation between these gene expression levels an...
Gene Expression Levels as Predictive Markers of Outcome in Pancreatic Cancer after Gemcitabine-Based Adjuvant Chemotherapy12
Neoplasia. 2010 October;12(10):807-817.Figure 5
Correlation between GEM score and survival time. DFS time (A) and OS time (B) after resection of PDAC categorized by combined GEM score (hENT1 score x dCK score x RRM1 score x RRM2 score) in GEM group patients. High GEM scores were well correlated with prolonged DFS time (A) and OS time (B). *P < .05.
Gene Expression Levels as Predictive Markers of Outcome in Pancreatic Cancer after Gemcitabine-Based Adjuvant Chemotherapy12
Neoplasia. 2010 October;12(10):807-817.Figure 6
Quantitative analyses of mRNA associated with gemcitabine sensitivity in EUS-FNA cytologic specimens. Representative micrographs of cytologic specimens obtained from patients with PDAC who underwent EUS-FNA cytologic examination (A, B). Most samples consisted of a large amount of blood and inflammat...
Gene Expression Levels as Predictive Markers of Outcome in Pancreatic Cancer after Gemcitabine-Based Adjuvant Chemotherapy12
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Abstract

Adenovirus-mediated gene therapy shows remarkable promise as a new strategy for advanced pancreatic cancer, but satisfactory clinical results have not yet been obtained. To improve this gene therapy, we investigated the effects of gemcitabine (GEM) on transgene expression by adenoviral vectors and their biological effects. We used Ad-lacZ and adenoviral vector-expressing NK4 (Ad-NK4) as representative adenoviral vectors. These vectors express beta-galactosidase (beta-gal) and NK4 (which inhibits the invasion of cancer cells), respectively, under the control of the CMV promoter. Cells were infected with the individual adenoviruses and then treated with GEM. GEM increased beta-gal mRNA expression and beta-gal activity, and increased NK4 expression in both culture media and within infected cells, in dose-dependent manners. The increased expression of NK4 delivered by Ad-NK4 had biological effects by inhibiting the invasion of cancer cells. GEM also enhanced NK4 expression in SUIT-2 cells transfected with an NK4-expressing plasmid, suggesting that GEM enhanced CMV promoter activity. In in vivo experiments, NK4 expression within subcutaneously implanted tumors was increased in GEM-treated mice compared with control mice. These results suggest that adenovirus-mediated gene therapy with GEM may be a promising approach for treating pancreatic cancer, and that this combination therapy may decrease the risks of side effects.

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BACKGROUND: Recently, cancer stem cells have been reported as a new therapeutic target in pancreatic cancer as well as other cancers, but the specific role of these cells is unknown. METHODS: The authors investigated the functional roles of CD133+ cells, 1 of the putative cancer stem cell candidates in pancreatic cancer. CD133 expression was assessed in human pancreatic cancer and cancer cell lines by quantitative real-time reverse transcriptase polymerase chain reaction and flow cytometry. Next, they compared the ability of CD133+ and CD133- cells to proliferate, migrate, and invade using 2 pancreatic cancer cell lines. In particular, they evaluated the relationship between CD133+ cells and primary pancreatic stromal cells. RESULTS: CD133 was expressed in primary human pancreatic cancer tissues and some cancer cell lines, whereas there was little expression in primary normal pancreatic epithelial cells and primary pancreatic stromal cells. CD133+ cells, isolated by flow cytometry, showed increased cell proliferation under anchorage-independent conditions (P<.01), and enhanced migration and invasion, particularly when cocultured with primary pancreatic stromal cells (P<.001). Chemokine-related receptor-4 (CXCR4), markedly overexpressed in CD133+ cells, may be responsible for the increased invasive ability of the cells cocultured with pancreatic stromal cells, which express stromal derived factor-1, the ligand to CXCR4. CONCLUSIONS: These data suggest that CD133+ cells exhibit more aggressive behavior, such as increased cell proliferation, migration, and invasion, especially in the presence of pancreatic stromal cells. The targeting therapy for the interaction between CD133+ cancer cells and stromal cells may be a new approach for the treatment of pancreatic cancer. Copyright (c) 2010 American Cancer Society.

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PURPOSE:Pediatric surgeons require highly advanced skills when performing endoscopic surgery, but their experience with such cases tend to be limited in comparison to general surgeons. The aim of this study was to evaluate the effectiveness of basic endoscopic surgery training for less-experienced young pediatric surgeons and then compare their skills with those of general surgeons.METHODS:The surgeons (n = 477) subjected to this study underwent a 2-day endoscopic skill training program, consisting of lectures, box training, VR simulator training, tissue training, and live tissue training. The trainees were divided into two groups: P (pediatric surgeons, n = 33) and G (general surgeons, n = 444). The trainees were required to make a continuous suture along a circle measuring 2.5 cm in diameter and the findings were evaluated both before and after training. A statistical analysis was conducted using the unpaired t test.RESULTS:The number of experienced
cases totaled 20.8 ± 23.9 in P and 60.6 ± 80.5 in G (p &LT; 0.001). The number of completed sutures before training was 1.4 ± 1.1 in P and 1.9 ± 1.5 in G (p &LT; 0.05). The number of completed sutures after training was 4.1 ± 1.3 in P and 3.9 ± 1.9 in G (p &GT; 0.05). The economy and speed of the forceps improved, however, the number of errors increased.CONCLUSION:Less-experienced pediatric surgeons improved their surgical skill and ability until reaching almost the same level as that observed in more experienced general surgeons during training, however, the number of errors after training increased in comparison to before training. As a result, this program needs to be modified to reduce the number of errors while enabling pediatric surgeons to master the safe and precise surgical techniques needed in this field.

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Abstract

To overcome the limited clinical efficacy of conditionally replicative adenoviruses (CRAds), we investigated the effects of combination therapy with gemcitabine (GEM) and the hTERT-promoter-dependent CRAd (hTERT-CRAd), Ad5/3hTERTE1. This combination therapy exhibited enhanced cytotoxic effects on pancreatic cancer both in vitro and in vivo. Furthermore, we revealed that this enhancement effect was due to the multiple bidirectional interactions between hTERT-CRAd and GEM. The GEM-sensitizing effect of E1 expression derived from hTERT-CRAd, and the enhancement effect by GEM on hTERT promoter activity which led to the increase of adenovirus E1 and viral infectivity. This combination therapy may be a promising therapeutic approach for pancreatic cancer.

Copyright 2010 Elsevier Ireland Ltd. All rights reserved.

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CD24 is a molecule involved in cell adhesion and tumor metastasis. The aims of this study were (1) to evaluate the association between CD24 expression and the progression of intraductal papillary mucinous neoplasms of the pancreas and (2) to investigate the association between CD24 expression in pancreatic cancer and the prognosis of patients who underwent curative pancreatectomy. Immunohistochemical analysis of CD24 was performed for 95 intraductal papillary mucinous neoplasms of the pancreas and 83 pancreatic cancers. We investigated the association between CD24 expression and the histologic grade of intraductal papillary mucinous neoplasms of the pancreas, the clinicopathologic parameters of pancreatic cancers, and the survival time of pancreatic cancer patients who underwent pancreatectomy. The positive rates of CD24 expression in intraductal papillary mucinous adenoma, borderline intraductal papillary mucinous neoplasm, noninvasive intraductal papillary mucinous carcinoma, and invasive intraductal papillary mucinous carcinoma were 5 (20%) of 24, 12 (48%) of 25, 10 (43%) of 23, and 15 (65%) of 23, respectively. The CD24-positive rates were significantly higher in borderline intraductal papillary mucinous neoplasm and intraductal papillary mucinous carcinoma compared with intraductal papillary mucinous adenoma (P = .046 and P = .007, respectively). The staining scores, which were determined from the percentage of stained cells and the staining intensity, were significantly higher in invasive intraductal papillary mucinous carcinoma than in noninvasive intraductal papillary mucinous carcinoma (P = .043). In the pancreatic cancers, higher tumor stage (P = .007), nodal metastasis (P = .021), and higher-grade tumors (P < .001) were more frequent in the CD24-positive group compared with the CD24-negative group. CD24 expression was associated with shorter survival in univariate analysis (P = .028) However, based on the multivariate analysis, the CD24 expression was not associated with survival. In conclusion, CD24 is involved in the progression of intraductal papillary mucinous neoplasms of the pancreas and in the malignant behavior of pancreatic cancers.

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OBJECTIVES: The purpose of this study was to examine
CD44v6 expression in intraductal papillary mucinous neoplasms (IPMNs)
and clarify the role of CD44v6 in progression, invasion, metastasis, and
morphogenesis of IPMNs. METHODS: One hundred fifty-one samples of IPMNs
and 30 normal controls were subjected to immunohistochemical analysis
for CD44v6. The IPMNs were divided into 4 groups according to the grade
of atypia (adenoma, borderline IPMN, noninvasive carcinoma, and invasive
carcinoma) and 5 subtypes according to histological phenotype (gastric,
intestinal, pancreatobiliary, oncocytic, and unclassified). Correlations
were investigated between CD44v6 expression and clinicopathological
characteristics including grade of atypia, subtype, lymph node
metastasis, and invasion pattern. RESULTS: Whereas normal ductal
epithelium did not express CD44v6, CD44v6 expression was observed from
the early stage of IPMNs and up-regulated in the progression of IPMNs
to invasive carcinoma. CD44v6 expression in intestinal-type IPMNs was
significantly lower compared with that in other subtypes. Whereas no
correlation was observed between lymph node metastasis and CD44v6
expression in invasive IPM carcinomas, the invasion pattern was
significantly correlated to CD44v6 expression. CONCLUSIONS: The present
data indicate that CD44v6 expression determines the morphology and
aggressiveness of IPMNs and is involved in development and invasion of
IPMNs.

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Abstract

Adenovirus-mediated gene therapy combined with chemotherapeutic agents is expected to represent a new approach for treating pancreatic cancer. However, there have been no reports of definitive effects of chemotherapeutic agents on adenovirus-mediated gene therapies. In the present study, we investigated the effects of chemotherapeutic agents on the transduction efficiency of an adenovirus-based gene therapy. Adenovirus (Ad-NK4) expressing NK4, which acts as a hepatocyte growth factor antagonist, was used as a representative gene therapy. Pancreatic cancer cells infected with Ad-NK4 were treated with chemotherapeutic agents (5-fluorouracil [5FU], cisplatin or etoposide), and the NK4 levels in their culture media were measured. To examine the effects of chemotherapeutic agents in vivo, Ad-NK4 was administered to subcutaneous tumors in mice after treatment with the agents, and the tumor NK4 levels were measured. The NK4 levels in culture media from cells treated with 5FU, cisplatin and etoposide were 5.2-fold (P = 0.026), 6-fold (P < 0.001) and 4.3-fold (P < 0.001) higher than those of untreated cells, respectively. The chemotherapeutic agents also increased Ad-NK4 uptake. The NK4 levels in tumors treated with 5FU, cisplatin and etoposide were 5.4-fold (P = 0.006), 11.8-fold (P < 0.001) and 4.9-fold (P = 0.017) higher than those in untreated tumors, respectively. The present findings suggest that chemotherapeutic agents significantly improve the efficiency of adenovirus-mediated gene transfer in pancreatic cancer. Furthermore, they will contribute to decreases in the adenovirus doses required for gene transfer, thereby controlling the side-effects of adenovirus infection in normal tissues.

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Adenovirus-mediated gene therapy is a promising approach for the treatment of pancreatic cancer. We previously reported that radiation enhanced adenovirus-mediated gene expression in pancreatic cancer, suggesting that adenoviral gene therapy might be more effective in radioresistant pancreatic cancer cells. In the present study, we compared the transduction efficiency of adenovirus-delivered genes in radiosensitive and radioresistant cells, and investigated the underlying mechanisms. We used an adenovirus expressing the hepatocyte growth factor antagonist, NK4 (Ad-NK4), as a representative gene therapy. We established two radioresistant human pancreatic cancer cell lines using fractionated irradiation. Radiosensitive and radioresistant pancreatic cancer cells were infected with Ad-NK4, and NK4 levels in the cells were measured. In order to investigate the mechanisms responsible for the differences in the transduction efficiency between these cells, we measured expression of the genes mediating adenovirus infection and endocytosis. The results revealed that NK4 levels in radioresistant cells were significantly lower (P < 0.01) than those in radiosensitive cells, although there were no significant differences in adenovirus uptake between radiosensitive cells and radioresistant cells. Integrin beta3 was up-regulated and the Coxsackie virus and adenovirus receptor was down-regulated in radioresistant cells, and inhibition of integrin beta3 promoted adenovirus gene transfer. These results suggest that inhibition of integrin beta3 in radioresistant pancreatic cancer cells could enhance adenovirus-mediated gene therapy.

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Pancreatic ductal adenocarcinoma is often characterized by an abundant desmoplastic stroma that is partially induced by activated pancreatic stellate cells (PSCs). Indirect co-culture has often been used to investigate the effects of cancer-stromal interactions on the proliferation of cancer cells, but the effects of cell-cell adhesion and juxtacrine signaling between cancer and stromal cells cannot be evaluated using this method. This study aimed to establish a simplified direct co-culture system that could be used to quantify populations of cancer cells in co-culture with PSCs, and to evaluate the effects of direct cell contact on the proliferation of cancer cells. We established three green fluorescent protein (GFP)-expressing pancreatic cancer cell lines and were able to quantify them with high reliability and reproducibility, even when co-cultured directly with PSCs, using a color plate reader. We assessed the differential effects of direct and indirect co-culture with PSCs on the proliferation of cancer cells, and found that the proliferation of GFP-expressing pancreatic cancer cell lines was dramatically enhanced by direct co-culture with PSCs, compared with the indirect co-culture system. We also found that direct co-culture of cancer cells and PSCs activated the Notch signaling pathway in both cell types. Direct cell contact between cancer cells and PSCs plays an important role in the control of cancer cell proliferation, and is essential to the understanding of tumor-stromal interactions

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BACKGROUND: To date, several training and evaluation systems for endoscopic surgery have been developed, such as virtual-reality simulators and box trainers. However, despite current advances in these objective assessments, no functional brain studies during learning of endoscopic surgical skills have been carried out. In the present study, we investigated cortical activation using near-infrared spectroscopy (NIRS) during endoscopic surgical tasks. STUDY DESIGN: A total of 21 right-handed subjects, comprising 4 surgical experts, 4 trainees, and 13 novices, participated in the study. Suturing and knot-tying tasks were performed in a box trainer. Cortical activation was assessed in all subjects by task-related changes in hemoglobin (Hb) oxygenation using NIRS. RESULTS: In surgical experts and novices with no experience of endoscopic surgical training, we found no changes in oxy-Hb, deoxy-Hb or total-Hb levels in any of the frontal channels. In surgical trainees and one novice with experience of endoscopic surgical training, we found significant increases in oxy-Hb and total-Hb levels in most of the frontal channels. There were significant differences in oxy-Hb and total-Hb levels in CH-19 between surgical experts and trainees (p = 0.02 for both), and between surgical trainees and novices with no experience of endoscopic surgical training (p = 0.008 for both). Furthermore, additional training increased oxy-Hb levels in the frontal cortex of novices with no experience of endoscopic surgical training but had no such effect on surgical experts. CONCLUSIONS: The present data suggest that NIRS is a feasible tool for assessing brain activation during endoscopic surgical tasks, and may have a large impact on the future development of teaching, training, and assessment methods for endoscopic surgical skills.

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BACKGROUND: Laparoscopic surgeons require extended experience of cases to overcome the lack of depth perception on a two-dimensional (2D) display. Although a three-dimensional (3D) display was reported to be useful over two decades ago, 3D systems have not been widely used. Recently, we developed a novel 3D dome-shaped display (3DD) system, CyberDome.

STUDY DESIGN: In the present study, a total of 23 students volunteered. We evaluated the effects of the 3DD system on depth perception and laparoscopic procedures in comparison with the 2D, a conventional 3D (3DP) or the 2D high definition (HD) systems using seven tasks.

RESULTS: The 3DD system significantly improved depth perception and laparoscopic performance compared with the 2D system in six new tasks. We further found that the 3DD system shortened the execution time and reduced the number of errors during suturing and knot tying. The 3DD system also provided more depth perception than the 3DP and 2D HD systems.

CONCLUSIONS: The novel 3DD system is a promising tool for providing depth perception with high resolution to laparoscopic surgeons.

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OBJECTIVE: The aim of this study is to evaluate the clinical significance of S100A4 mRNA expression in pancreatic cancer. MATERIALS AND METHODS: We obtained invasive ductal carcinoma (IDC) cells from ten lesions, intraductal papillary mucinous neoplasm (IPMN) cells from 20 lesions, and normal ductal cells from 20 normal pancreatic tissues by laser microdissection of frozen tissues. S100A4 expression was examined in the microdissected cells and in formalin-fixed paraffin-embedded (FFPE) samples of 87 pancreatic cancers by quantitative reverse transcription-polymerase chain reaction. RESULTS: IDC cells expressed higher levels of S100A4 than IPMN cells (P = 0.002) and normal ductal cells (P < 0.001), although the difference between IPMN cells and normal ductal cells was not statistically significant (P = 0.070). Analysis of FFPE samples revealed that high S100A4 expression was significantly associated with a shorter overall survival (P = 0.023). In immunohistochemical analysis, the extent of S100A4 mRNA expression was significantly correlated with the expression of S100A4 protein (P = 0.028). CONCLUSION: S100A4 could be a marker for malignancy in pancreatic tumors and for poor prognosis in patients with pancreatic cancer.

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Subclassification of intraductal papillary mucinous neoplasms of the pancreas (IPMNs), based on morphological features and immunohistochemical profiles, has been proposed. Intestinal-type IPMNs frequently show moderate to severe dysplasia. Regenerating islet-derived family, member 4 (REG4) is associated with the adenoma-carcinoma sequence in colon cancer and it is also associated with intestinal phenotype. Therefore, to identify REG4 expression in IPMNs may be helpful to detect high-grade IPMNs. We also investigated REG4 expression and CDX2 expression in IPMNs. To investigate the expressions of REG4 and CDX2 in IPMNs and in invasive ductal adenocarcinoma derived from IPMN, we used immunohistochemical staining and microdissection-based quantitative real-time reverse transcription-polymerase chain reaction. Among 125 IPMNs, 43 (34%) were positive for REG4 and most of the intestinal-type IPMNs showed its expression (35/38). The positive ratio of REG4 expression in colloid carcinoma (5/7) was significantly higher than that in tubular carcinoma (1/17; P=0.003). Most of CDX2-positive cases (31/33) expressed REG4 protein, whereas only 12 of 92 CDX2-negative cases did (P<0.001). The levels of REG4 mRNA in intestinal-type IPMN were significantly higher compared to those in gastric-type IPMN or to normal pancreatic ductal epithelium (P=0.005, P=0.004, respectively). REG4 expression was observed more frequently in borderline lesions (14/28) and carcinoma (21/45) compared to adenoma (8/52). Using the Ki-67 labeling index, REG4 expression was significantly correlated with proliferative activity in borderline lesions. We conclude that REG4 is involved in the 'intestinal' pathway of carcinogenesis in IPMN.Modern Pathology advance online publication, 9 January 2009; doi:10.1038/modpathol.2008.205.

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Abstract

Due to the poor prognosis of pancreatic cancer, novel diagnostic modalities for early diagnosis and new therapeutic strategy are urgently needed. Recently, microRNA-21 (miR-21) was reported to be strongly overexpressed in pancreatic cancer as well as in other solid cancers. We investigated the functional roles of miR-21, which have not been fully elucidated in pancreatic cancer. miR-21 expression was assessed in pancreatic cancer cell lines (14 cancer cell lines, primary cultures of normal pancreatic epithelial cells and fibroblasts, and a human normal pancreatic ductal epithelial cell line) and pancreatic tissue samples (25 cancer tissues and 25 normal tissues) by quantitative real-time reverse transcription-PCR amplification. Moreover, we investigated the proliferation, invasion, and chemoresistance of pancreatic cancer cells transfected with miR-21 precursor or inhibitor. miR-21 was markedly overexpressed in pancreatic cancer cells compared with nonmalignant cells, and miR-21 in cancer tissues was much higher than in nonmalignant tissues. The cancer cells transfected with the miR-21 precursor showed significantly increased proliferation, Matrigel invasion, and chemoresistance for gemcitabine compared with the control cells. In contrast, inhibition of miR-21 decreased proliferation, Matrigel invasion, and chemoresistance for gemcitabine. Moreover, miR-21 positively correlated with the mRNA expression of invasion-related genes, matrix metalloproteinase-2 and -9, and vascular endothelial growth factor. These data suggest that miR-21 expression is increased in pancreatic cancer cells and that miR-21 contributes to the cell proliferation, invasion, and chemoresistance of pancreatic cancer.

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PURPOSE: LIM domain only 2 (LMO2) has been identified as a novel oncogene associated with carcinogenesisand better prognosis in several malignant tumors. We investigate the involvement of LMO2 in pancreatic cancer.EXPERIMENTAL DESIGN: We evaluated LMO2 expression in cultured cells, bulk tissues, and microdissected cellsfrom pancreatic cancers by quantitative reverse transcription朴olymerase chain reaction and immunohistochemistry.RESULTS: Of 164 pancreatic cancers, 98 (60%) were positive for LMO2 expression. LMO2 was more frequently detectedin high-grade pancreatic intraepithelial neoplasia (PanIN) lesions (PanIN-2 and -3) than in low-grade PanIN lesions(PanIN-1A and -1B; P &LT; .001) and was not detected in normal pancreatic ductal epithelium. The LMO2messenger RNA levels were significantly higher in invasive ductal carcinoma cells than in normal pancreatic cellsas evaluated by quantitative reverse transcription朴olymerase chain reaction
a!
nalyses of microdissected cells (P =.036). We also found higher incidence of LMO2 expression in histologic grade G1/G2 cancers than in grade G3cancers (P &LT; .001). The median survival time of LMO2-positive patients was significantly longer than that ofLMO2-negative patients (P &LT; .001), and multivariate analyses revealed that high LMO2 expression was an independentpredictor of longer survival (risk ratio, 0.432, P &LT; .001). Even among patients with a positive operative margin,LMO2-positive patients had a significant survival benefit compared with LMO2-negative patients. We furtherperformed a large cohort study (n = 113) to examine the LMO2 messenger RNA levels in formalin-fixed paraffinembeddedsamples and found similar results. CONCLUSIONS: LMO2 is a promising marker for predicting a betterprognosis in pancreatic cancer.Neoplasia (2009) 11, 712・19IntroductionPancreatic cancer is the fourth leading cause of cancer-related deathin Western countries and has the lowest pat
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ent survival rate of anysolid cancer [1・]. Recently, although the cancer death rates of mostmalignancies have decreased owing to improvements in early detectionand treatment, the overall 5-year survival of patients with pancreaticcancer has only slightly increased from 3% to 5% [1] becauseof difficulties in the diagnosis of pancreatic cancer at early stages. Surgicalresection is the only curative treatment of pancreatic cancer, andthe survival rate for patients with a negative operative margin status(R0) is significantly higher than that for patients with positive operativemargin status (R1 and R2) [4]. However, some patients with apositive operative

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AbstractLaparoscopic gastrectomy is widely used as minimally invasive surgery for gastric carcinoma. Billroth I or Roux-en-Y reconstruction is commonly performed after laparoscopic distal gastrectomy (LDG). Roux-en-Y reconstruction after LDG is one of the best methods for reconstruction of the alimentary tract when Billroth I reconstruction is difficult. There are few reports of intracorporeal Roux-en-Y reconstruction after LDG because of the technical difficulties of such a procedure. In particular, in the case of a very small gastric remnant, gastrojejunostomy using endoscopic linear staplers becomes more complicated. We developed a new technique for intracorporeal Roux-en-Y reconstruction: a modified stapling technique to allow the gastrojejunostomy to be made on the stomach transecting line that is applicable even when the residual stomach is very small. Roux-en-Y reconstruction with our modified technique was performed in six patients. There was
no intraoperative complication or conversion to minilaparotomy or conventional celiotomy in any patient. Oral intake was easy and adequate after surgery. The present Roux-en-Y reconstruction procedure is feasible. Herein we describe an intraabdominal Roux-en-Y reconstruction with a modified stapling technique after LDG.

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BACKGROUND: Laparoscopic surgeons require extended experience of cases to overcome the lack of depth perception on a two-dimensional (2D) display. Although a three-dimensional (3D) display was reported to be useful over two decades ago, 3D systems have not been widely used. Recently, we developed a novel 3D dome-shaped display (3DD) system, CyberDome. STUDY DESIGN: In the present study, a total of 23 students volunteered. We evaluated the effects of the 3DD system on depth perception and laparoscopic procedures in comparison with the 2D, a conventional 3D (3DP) or the 2D high definition (HD) systems using seven tasks. RESULTS: The 3DD system significantly improved depth perception and laparoscopic performance compared with the 2D system in six new tasks. We further found that the 3DD system shortened the execution time and reduced the number of errors during suturing and knot tying. The 3DD system also provided more depth perception than the 3DP and 2D HD systems. CONCLUSIONS: The novel 3DD system is a promising tool for providing depth perception with high resolution to laparoscopic surgeons.

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BACKGROUND: The functional roles of deoxycytidine kinase (dCK) in acquired resistance to gemcitabine remain unknown in pancreatic cancer. Here, the functional involvement of dCK in gemcitabine-resistance of pancreatic cancer was investigated. MATERIALS AND METHODS: The levels of the dCK gene as well as other gemcitabine-related genes (hENT1, RRM1 and RRM2) were analyzed in gemcitabine-resistant pancreatic cancer cells (GR cells) using quantitative real-time reverse transcription polymerase chain reaction. The effects of inhibition of these genes on sensitivity to gemcitabine were evaluated. RESULTS: In GR cells, expression of dCK was significantly reduced compared with that of parental cells (p < 0.05). The dCK-targeting siRNA significantly reduced gemcitabine sensitivity (p < 0.01) without affecting cell proliferation. The RRM1- and RRM2-targeting siRNAs increased gemcitabine sensitivity (p < 0.05) and reduced cell proliferation even without gemcitabine treatment. The hENT-targeting siRNA did not affect gemcitabine sensitivity or cell proliferation. CONCLUSION: Down-regulation of dCK specifically enhanced acquired resistance

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PURPOSE: Proliferative diabetic retinopathy (PDR) results from the formation of fibrovascular membranes (FVMs) in the posterior fundus that can lead to a severe decrease of vision. Tumor endothelial marker 7 (TEM7) is a protein that is highly expressed in the endothelial cells of tumors, but whether it plays a role in FVMs is unknown. The purpose of this study was to determine whether TEM7 is associated with the formation of FVMs. METHODS: FVMs were obtained during vitrectomy from patients with PDR. RT-PCR was performed to determine the level of expression of the mRNA of TEM7. The splice variants of TEM7 were identified by direct sequencing. Immunohistochemical analyses and in situ hybridization was performed to determine the sites of TEM7 in the FVMs. RESULTS: The level of the mRNA of TEM7 was high in 10 of 10 FVMs but was barely detectable in the five idiopathic epiretinal membranes. Direct sequencing of subcloned TEM7 PCR products revealed several splice variants (intracellular, secreted, and membrane-bound forms of TEM7) in the FVMs. Immunohistochemical analysis showed a colocalization of TEM7 and CD34, an endothelial cell marker, in most of the neovascular endothelial cells in the FVMs. Immunoelectron microscopy revealed that membrane-bound TEM7 was expressed on the luminal surfaces of the vascular endothelial cells of FVMs. CONCLUSIONS: This study indicates that TEM7 may play a significant role in the proliferation and maintenance of neovascular endothelial cells in the FVMs. If correct, TEM7 may be a molecular target for new diagnostic and therapeutic strategies for PDR.

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PURPOSE: Adenovirus-mediated gene therapy combined with radiation is expected to be a new approach to treat pancreatic cancer. However, there are no reports of definitive effects of radiation on adenovirus-mediated gene therapies. In the present study, we investigated the effect of radiation on the transduction efficiency of an adenovirus-based gene therapy. EXPERIMENTAL DESIGN: We used adenovirus expressing NK4 (Ad-NK4), an antagonist for hepatocyte growth factor, as a representative gene therapy. Pancreatic cancer cells preinfected with Ad-NK4 were irradiated, and NK4 levels in culture media of these cells were measured. We investigated cytomegalovirus (CMV) promoter activity and uptake of adenovirus in these cells. To examine the effect of radiation in vivo, Ad-NK4 was given to irradiated subcutaneous tumors in nude mice, and NK4 levels in tumors were measured. RESULTS: NK4 levels in culture media of irradiated cells were 4.5-fold (P < 0.01) higher than those of nonirradiated cells. Radiation enhanced activation of the CMV promoter and adenovirus uptake (P < 0.01), leading to increased levels of NK4. We found that activation of p38 mitogen-activated protein kinase and up-regulation of dynamin 2 may be involved in the radiation-induced activation of the CMV promoter and adenovirus uptake, respectively. NK4 levels in irradiated tumors were 5.8-fold (P = 0.017) higher than those in nonirradiated tumors. CONCLUSIONS: The present findings suggest that radiation significantly improves the efficiency of adenovirus-mediated gene transfer in pancreatic cancer and probably contributes to decreasing the dose of adenovirus required for gene transfer and controlling side effects of adenovirus infection in nonirradiated normal tissue.

PMID: 18347189 [PubMed - indexed for MEDLINE]

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Clinicians frequently require cytopathological assessment of tumor samples for preoperative diagnosis, but in some specimens, diagnosis remains inconclusive after cytological examination. To date, several molecular markers, including human telomerase reverse transcriptase (hTERT), have been assessed for the ability to detect malignancy. However, analyses using whole cytological specimens are generally affected by contamination of untargeted cells. The present study investigated the feasibility of more sensitive examination by quantitative mRNA analysis of target cells microdissected from cytological specimens. Laser capture microdissection (LCM) was used to obtain target cells from cytological specimens. hTERT mRNA levels were then measured in target cells by quantitative real-time RT-PCR (qRT-PCR). The effect of RNA fragmentation on qRT-PCR was also assessed. Total RNA from cytological specimens was sometimes fragmented to a large degree. To avoid the effect of RNA fragmentation, gene specific priming and PCR primers generating short PCR products were used and no difference in delta Ct values between fragmented and non-fragmented RNA were found. hTERT mRNA levels were measured in cells microdissected from 33 cytological specimens. The levels of hTERT mRNA were significantly higher in malignant cases compared to those in non-malignant cases (P = 0.0003). The sensitivity was 96.2%, even when the specificities were 100%. High levels of hTERT mRNA were also found in three cases that were not diagnosed as malignant by cytological examination. Quantitative assessment of hTERT mRNA levels in cells microdissected from cytological specimens is a potential diagnostic tool to potentiate cytological examination in diagnosing malignancy.

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Purpose Midkine (MK) has been reported to be a possible molecular marker for the diagnosis of pancreatic cancer. We investigated the feasibility of quantitative analysis of MK mRNA by quantitative real-time RT-PCR (qRT-PCR) as a promising tool for the diagnosis of pancreatic cancer. Results We found that pancreatic cancer tissues expressed significantly higher levels of MK mRNA than intraductal pancreatic mucinous neoplasm (IPMN) and non-neoplastic pancreatic tissues (P < 0.05); in contrast, we did not find any differences in MK mRNA expression between IPMN and non-neoplastic pancreatic tissues. Additionally, we observed that poorly differentiated carcinoma samples expressed higher levels of MK mRNA than well-differentiated carcinoma samples, although a significant difference was not observed. Conclusions The present data suggests that quantitative analysis of MK mRNA provides an objective and sensitive evaluation and may be a promising modality for the diagnosis of pancreatic cancer and the prediction of its prognosis.

PMID: 18712601 [PubMed - as supplied by publisher]

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BACKGROUND: LIM-only 4 (LMO4), a member of the LIM-only (LMO) subfamily of LIM domain-containing transcription factors, was initially reported to have an oncogenic role in breast cancer. We hypothesized that LMO4 may be related to pancreatic carcinogenesis as it is in breast carcinogenesis. If so, this could result in a better understanding of tumorigenesis in pancreatic cancer. METHODS: We measured LMO4 mRNA levels in cultured cells, pancreatic bulk tissues and microdissected target cells (normal ductal cells; pancreatic intraepithelial neoplasia-1B [PanIN-1B] cells; PanIN-2 cells; invasive ductal carcinoma [IDC] cells; intraductal papillary-mucinous adenoma [IPMA] cells; IPM borderline [IPMB] cells; and invasive and non-invasive IPM carcinoma [IPMC]) by quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR). RESULTS: 9 of 14 pancreatic cancer cell lines expressed higher levels of LMO4 mRNA than did the human pancreatic ductal epithelial cell line (HPDE). In bulk tissue samples, expression of LMO4 was higher in pancreatic carcinoma than in intraductal papillary-mucinous neoplasm (IPMN) or non-neoplastic pancreas (p < 0.0001 for both). We carried out microdissection-based analyses. IDC cells expressed significantly higher levels of LMO4 than did normal ductal epithelia or PanIN-1B cells (p < 0.001 for both) or PanIN-2 cells (p = 0.014). IPMC cells expressed significantly higher levels of LMO4 than did normal ductal epithelia (p < 0.001), IPMA (p < 0.001) and IPMB cells (p = 0.003). CONCLUSION: Pancreatic carcinomas (both IDC and IPMC) expressed significantly higher levels of LMO4 mRNA than did normal ductal epithelia, PanIN-1B, PanIN-2, IPMA and IPMB. These results suggested that LMO4 is overexpressed at late stages in carcinogenesis of pancreatic cancer.

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The hedgehog (Hh) signaling pathway has been reported to be associated with the growth of pancreatic cancer, but its role in the invasive phenotype is poorly understood. Therefore, we investigated the role of the Hh pathway in pancreatic cancer cell invasiveness using a Matrigel invasion assay. Blockade of the Hh pathway by cyclopamine inhibited pancreatic cancer cell invasion in association with a decreased expression of matrix metalloproteinase (MMP)-9. By contrast, activation of the Hh pathway by the addition of exogenous Sonic hedgehog increased cell invasion and MMP-9 expression. Stable transfection of pancreatic cancer cells with Gli1 increased their invasiveness, which was associated with activation of MMP-9. We also showed that inhibition of MMP-9 by small interfering RNA blocked the increased invasiveness of Gli1-transfected cells. Furthermore, inhibition of Gli1 by small interfering RNA suppressed the invasiveness and MMP-9 expression of pancreatic cancer cells. Taken together, these findings suggest that members of the Hh pathway, especially Gli1, play an important role in the invasiveness of pancreatic cancer cells through the regulation of MMP-9 expression.

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Intraductal papillary mucinous neoplasm (IPMN) is a well-established entity in pancreatic neoplasms and a precursor of infiltrating adenocarcinoma. Fascin, an actin-bundling protein involved in cellular motility, is upregulated in many human neoplasms. Its overexpression in pancreatic intraepithelial neoplasia, a pre-cancerous lesion sharing many characteristics with IPMN, has been reported. However, fascin expression in IPMN remains unknown. The aim of this study was to investigate fascin expression in IPMNs and to elucidate its relationship to clinicopathological features, including histological grade and phenotypic subclassification. We evaluated fascin expression by immunohistochemistry in 116 surgical specimens, followed by quantitative analysis of fascin mRNA expression using a laser microdissection system and real-time reverse-transcriptase polymerase chain reaction in eight frozen samples. Fascin expression was significantly higher in borderline neoplasms (25/29, 86%) and carcinomas (37/42, 88%) than in adenomas (23/45, 51%) (P<0.05, respectively), but no difference was observed between borderline neoplasms and carcinomas. With regard to the subclassification, intestinal-type neoplasms (35/39, 90%) were more frequently positive for fascin than gastric-type neoplasms (36/59, 61%) (P<0.05). Two oncocytic-type neoplasms were both fascin-negative. Fascin mRNA expression seemed to be higher in moderately to severely dysplastic epithelium than in mildly dysplastic epithelium (not statistically significant), supporting the immunohistochemical experiments. Our findings suggest that fascin overexpression is involved in the progression of IPMN. Fascin could become a new therapeutic target for inhibition of their progression.Modern Pathology (2007) 20, 552-561. doi:10.1038/modpathol.3800763; published online 30 March 2007

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Despite evidence that Twist, a highly conserved basic helix-loop-helix transcription factor, is a novel oncogene, there are no reports describing Twist expression in pancreatic cancer. Intraductal papillary mucinous neoplasm (IPMN) and pancreatic intraepithelial neoplasia (PanIN) are precursor lesions of pancreatic cancer. To clarify involvement of Twist expression in pancreatic cancer, we used quantitative reverse transcription-polymerase chain reaction and examined Twist expression in pancreatic cancer, IPMN, and non-neoplastic pancreas using bulk tissues (11 cancers, 18 IPMNs, and 15 non-neoplastic pancreata), microdissected cells (cancer from 22 sections, IPMN from 19 sections, PanIN from 6 sections, and pancreatitis-affected epithelial cells from 14 sections), and pancreatic juice (16 from cancer, 28 from IPMN, and 17 from pancreatitis). Twist expression differed significantly between cancer and IPMN bulk tissues (p < 0.0001) but not between cancer and non-neoplastic tissues. Twist expressions differed significantly between microdissected cancer cells, IPMN cells, and pancreatitis-affected cells (all comparisons, p < 0.017). PanIN cells expressed significantly lower levels of Twist than did IDC cells (p = 0.016). Twist expression differed significantly between cancer and IPMN juice samples (p = 0.0002) but not between cancer and pancreatitis juice samples. Receiver operation characteristic curve analyses revealed that measurement of Twist was more useful for discriminating cancer from IPMN than from chronic pancreatitis (p = 0.009). Our results suggest that Twist is involved in tumor progression of pancreatic cancer and that measurement of Twist in pancreatic juice may be useful to differentiate pancreatic cancer from nonmalignant neoplasms such as IPMN. (c) 2007 Wiley-Liss, Inc.

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PURPOSE: Intraductal papillary mucinous neoplasms (IPMN) are known to show a transition from adenoma to carcinoma accompanied by several molecular abnormalities. ATM-Chk2-p53 DNA damage checkpoint activation, which is involved in prevention of the progression of several tumors, was analyzed to evaluate the role of the DNA damage checkpoint in the progression of IPMNs. EXPERIMENTAL DESIGN: One hundred and twenty-eight IPMNs were classified into four groups (intraductal papillary mucinous adenoma, borderline IPMN, noninvasive intraductal papillary mucinous carcinoma, and invasive intraductal papillary mucinous carcinoma) and stained immunohistochemically using antibody for Thr(68)-phosphorylated Chk2. Expression of ATM, Chk2, and p21(WAF1) and accumulation of p53 were also analyzed. RESULTS: Chk2 phosphorylation was shown in all adenomas and showed a significant decreasing trend with the progression of atypia (P < 0.0001 by the Cochran-Armitage test for trend). Expression of p21(WAF1) also exhibited a decreasing tendency (P < 0.0001), reflecting DNA damage checkpoint inactivation. p53 accumulation was mostly detected in malignant IPMNs. It was suggested that the DNA damage checkpoint provides a selective pressure for p53 mutation. CONCLUSION: Our findings indicate that DNA damage checkpoint activation occurs in the early stage of IPMNs and prevents their progression. It is suggested that disturbance of the DNA damage checkpoint pathway due to Chk2 inactivation or p53 mutation contributes to the carcinogenesis of IPMNs.

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There are few reports describing the diagnostic significance of S100A6 expression in clinical samples obtained from patients with pancreatic disease. In the present study, we measured S100A6 expression in pancreatic tissues and juice to evaluate its involvement in pancreatic carcinogenesis. We did quantitative real-time reverse transcription-PCR to measure mRNA expression in microdissected cells and pancreatic juice samples. Microdissected invasive ductal carcinoma and intraductal papillary mucinous neoplasm (IPMN) cells expressed significantly higher levels of S100A6 than did microdissected pancreatitis-affected epithelial and normal cells (all comparison; P < 0.008). Median levels of S100A6 in invasive ductal carcinoma were higher than those in IPMN, and those in pancreatitis-affected epithelial cells tended to be higher than those in normal cells, although these differences were not statistically significant. In analyses of pancreatic juice, IPMN and pancreatic cancer samples expressed significantly higher levels of S100A6 than did chronic pancreatitis samples (both; P < 0.017), but levels in pancreatic cancer and IPMN samples did not differ form each other. Receiver operating characteristic (ROC) curve analysis revealed that measurement of S100A6 was useful for discriminating cancer (area under the ROC curve, 0.864) or IPMN (area under the ROC curve, 0.749) from chronic pancreatitis. The present data suggest that expression of S100A6 is increased in a stepwise manner during pancreatic carcinogenesis and may be a biomarker for evaluating malignant potential. Measurement of S100A6 in pancreatic juice may be useful to detect early pancreatic cancer or identify individuals with high-risk lesions that may progress to pancreatic cancer.

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Controversy exists regarding the clinical significance of S100A2 in the progression of tumours. In pancreatic cancer, little is known about the role of S100A2. The aim of this study was to clarify the clinical significance of S100A2 expression in pancreatic carcinogenesis. We microdissected invasive ductal carcinoma (IDC) cells from 22 lesions, pancreatic intraepithelial neoplasia (PanIN) cells from five lesions, intraductal papillary mucinous neoplasm (IPMN) cells from 38 lesions, pancreatitis-affected epithelial (PAE) cells from 16 lesions, and normal ductal cells from 18 normal pancreatic tissues. S100A2 expression in 14 pancreatic cancer cell lines, microdissected cells and formalin-fixed paraffin-embedded (FFPE) samples was examined by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Microdissection analyses revealed that IDC cells expressed higher levels of S100A2 than did IPMN, PAE or normal cells (all comparisons, p < 0.007). Cell lines from metastatic sites expressed higher levels of S100A2 than those from primary sites. PanIN cells expressed higher levels of S100A2 than normal cells (p = 0.018). IDC cells associated with poorly differentiated adenocarcinoma expressed higher levels of S100A2 than did IDC cells without poorly differentiated adenocarcinoma (p = 0.006). Analyses of FFPE samples revealed that levels of S100A2 were higher in samples from patients who survived < 1000 days after surgery than in those from patients who survived > 1000 days (p = 0.043). Immunohistochemical analysis was consistent with qRT-PCR. S100A2 may be a marker of tumour progression or prognosis in pancreatic carcinogenesis and pancreatic cancer. Copyright 2007 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

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BACKGROUND: A disintegrin and metalloproteases (ADAMs) comprise a multifunctional family of membrane-anchored proteins. ADAM 9 and ADAM 15 are involved in cell migration and invasion. Expression of ADAM 9 and ADAM 15 was reported to be altered in several types of cancer. MATERIALS AND METHODS: Quantitative real-time reverse transcription-polymerase chain reaction was performed to measure the expression of ADAM 9 mRNA in bulk pancreatic tissues. Results showed no significant difference in the expression of ADAM 9 mRNA between pancreatic cancer and non-neoplastic pancreas. Primary cultured pancreatic fibroblasts also expressed ADAM 9 mRNA. Therefore, a laser microdissection and pressure catapulting technique was employed to isolate cancer cells from tumor tissues. The expression of ADAM 9 and ADAM 15 mRNA was measured in microdissected samples (cancer cells, n = 11; normal epithelial cells, n = 13 for ADAM 9; cancer cells, n = 9; normal epithelial cells, n = 9 for ADAM 15). RESULTS: Pancreatic cancer cells expressed significantly higher levels of ADAM 9 and ADAM 15 mRNA than did normal pancreatic epithelial cells (p = 0.016 for ADAM 9; p = 0.004 for ADAM 15). CONCLUSION: ADAM 9 and ADAM 15 are involved in pancreatic cancer. Microdissection-based analysis appears to be indispensable for the accurate analysis of the expression of certain ADAM family members in pancreatic cancer.

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To investigate c-met expression during early pancreatic carcinogenesis. METHODS: We used 46 bulk tissues and 36 micro-dissected samples, including normal pancreas, chronic pancreatitis, and pancreatic cancer, for quantitative real-time reverse transcription-polymerase chain reaction. RESULTS: In bulk tissue analyses, pancreatic cancer tissues expressed significantly higher levels of c-met than did chronic pancreatitis and normal pancreas tissues. c-met levels did not differ between chronic pancreatitis and normal pancreas tissues. In microdissection-based analyses, c-met was expressed at higher levels in microdissected pancreatic cancer cells and pancreatitis-affected epithelial cells than in normal ductal epithelial cells (both, P < 0.01). Interestingly, pancreatitis-affected epithelial cells expressed levels of c-met similar to those of pancreatic cancer cells. CONCLUSION: Overexpression of c-met occurs during the early stage of pancreatic carcinogenesis, and a single alteration of c-met expression is not sufficient for progression of chronic pancreatitis-affected epithelial cells to pancreatic cancer cells.

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Abstract
BACKGROUND:
Pancreatic cancer frequently recurs after operative treatment, resulting in a poor prognosis. Inhibition of proliferation of residual cancer cells is important for improved survival of patients with pancreatic cancer. Fibrin glue (FG) is a biocompatible, adherent hemostat that can deliver high concentrations of anticancer drugs to residual cancer cells. The aim of this study was to evaluate the local antitumor effect of a mixture of gemcitabine (GEM) and FG on pancreatic cancer cells implanted orthotopically in nude mice.
METHODS:
SUIT-2 human pancreatic cells were injected into the tail of the pancreas of nude mice. Seven days later, groups of mice were treated with 80 mg/kg GEM mixed with 0.5 mL fibrin glue (GEM + FG), 0.5 mL FG alone (FG), single intraperitoneal (i.p.) injection of 80 mg/kg GEM (GEM1), i.p. injection of 80 mg/kg GEM weekly for 3 weeks (GEM1,2,3), GEM + FG followed by weekly GEM injections for 2 weeks (GEM + FG + GEM2,3), or i.p. injection of PBS weekly for 3 weeks (controls).
RESULTS:
Twenty-eight days after cell injections, tumor volumes of groups treated with GEM + FG + GEM2,3, GEM1,2,3, GEM + FG, GEM1, and FG were decreased by 84%, 70%, 62%, 37%, and 10%, respectively, compared to that of control mice. GEM + FG + GEM2,3 had the strongest anticancer effect compared to all other groups (P < .05). Additionally, GEM + FG showed a more potent antitumor effect compared to GEM1 (P < .05). Survival of mice treated with GEM + FG + GEM2,3 was longer than that of mice in all other groups (P < .05).
CONCLUSIONS:
A mixture of GEM and FG was effective in inhibiting the growth of orthotopically implanted pancreatic neoplasms in nude mice. This procedure may be useful clinically to prevent the local recurrence of pancreatic cancer after pancreatectomy.

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Intraductal papillary mucinous neoplasms (IPMNs) are common cystic tumours of the pancreas. Sonic hedgehog (SHH) is involved in gastric epithelial differentiation and pancreatic carcinogenesis. However, a comprehensive analysis of SHH expression in IPMN has not yet been performed. In the present study, one-step quantitative real-time reverse transcription-polymerase chain reaction with gene-specific priming was used to examine mRNA levels in various types of clinical samples. SHH expression in IPMN was measured and the possible association of gastric epithelial differentiation with development of IPMN was evaluated. In bulk tissue analyses (IPMNs, 11 pancreatic cancer, and 20 normal pancreatic tissues), IPMN expressed significantly higher levels of SHH than did normal pancreas (IPMN versus normal pancreas, p = 0.0025; pancreatic cancer versus normal pancreas, p = 0.0132), but SHH expression did not differ between IPMN and pancreatic cancer (p = 0.3409). In microdissection analyses (infiltrating ductal carcinoma cells from 20 sections, IPMN cells from 20 sections, pancreatitis-affected epithelial cells from 11 sections, and normal epithelial cells from 12 sections), IPMN cells expressed significantly higher levels of SHH than did cancer cells, normal cells, or pancreatitis-affected ductal cells (all comparisons, p < 0.008). Pancreatic juice analyses (20 samples from pancreatic cancers, 31 samples from IPMNs, and 27 samples from chronic pancreatitis) revealed that SHH expression differed significantly between IPMN juice and pancreatitis juice (p < 0.0001), and between cancer juice and pancreatitis juice (p = 0.0125). Receiver operating characteristic curve analyses revealed that SHH measurement in pancreatic juice was useful for discriminating IPMN from chronic pancreatitis (area under the curve = 0.915; 95% confidence interval: 0.796-0.976). The data suggest that overexpression of SHH is an early event in the development of IPMN and that SHH measurement in pancreatic juice may provide some advantages for the treatment or follow-up of a subset of patients with IPMN or chronic pancreatitis.

Copyright (c) 2006 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

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PURPOSE: Our goal was to clarify the involvement and clinical significance of S100P in pancreatic carcinogenesis. EXPERIMENTAL DESIGN: We examined S100P expression in 45 bulk pancreatic tissues; in microdissected cells, including invasive ductal carcinoma (IDC) cells (20 sections), pancreatic intraepithelial neoplasia (PanIN) cells (12 sections), intraductal papillary mucinous neoplasm (IPMN) cells (19 sections), and normal epithelial cells (11 sections); and in pancreatic juice samples from 99 patients with pancreatic diseases (32 cancer, 35 IPMN, and 32 chronic pancreatitis samples). We used quantitative real-time reverse transcription-PCR with gene-specific priming to measure S100P in these various types of samples. RESULTS: In bulk tissue analyses, pancreatic cancer and IPMN expressed significantly higher levels of S100P than did nonneoplastic pancreas (P<0.017 and P=0.0013, respectively). Microdissection analyses revealed that IPMN expressed significantly higher levels of S100P than did IDC (P<0.0001) and PanIN (P=0.0031), although S100P expression did not differ between IDC and PanIN (P=0.077). In pancreatic juice analyses, cancer and IPMN juice expressed significantly higher levels of S100P than did pancreatitis juice (both P<0.0001). Receiver operating characteristic curve analyses revealed that measurement of S100P in pancreatic juice was useful for discriminating neoplastic disease from chronic pancreatitis (area under the curve=0.837; 95% confidence interval, 0.749-0.903). CONCLUSION: S100P may be an early developmental marker of pancreatic carcinogenesis, and measurement of S100P in pancreatic juice may be useful for early detection of pancreatic cancer or screening of early pancreatic carcinogenesis.

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Abstract

PURPOSE:

Recent microarray analyses revealed that expression of S100A11 is up-regulated in pancreatic cancer. The aim of the present study was to evaluate the association of S100A11 with pancreatic carcinogenesis.

EXPERIMENTAL DESIGN:

We measured S100A11 mRNA expression in various clinical samples related to pancreatic cancer and its precursor lesions, intraductal papillary mucinous neoplasm (IPMN) and pancreatic intraepithelial neoplasia, by quantitative reverse transcription-PCR.

RESULTS:

Levels of S100A11 were significantly higher in pancreatic cancer (n=22) and IPMN (n=18) bulk tissues than in nonneoplastic bulk tissues (n=22; P<0.0001 for both). Levels of S100A11 did not differ between pancreatic cancer and IPMN bulk tissues. In microdissection analyses, however, IPMN cells (n=21) expressed significantly higher levels of S100A11 than did cancer cells (n=23; P=0.003). The median level of S100A11 expression was higher in pancreatic intraepithelial neoplasia cells (n=6) than in cancer cells. In pancreatic juice analyses, cancer-related (n=24; P=0.004) and IPMN-related (n=18; P=0.001) juice expressed significantly higher levels of S100A11 than did chronic pancreatitis-related juice (n=23).

CONCLUSIONS:

The present data suggest that expression of S100A11, a putative tumor suppressor gene, is increased in the early stage of pancreatic carcinogenesis and decreased during subsequent progression to cancer. Analysis of the S100A11 level in pancreatic juice may be an effective tool for screening of patients with high-risk lesions that could progress to pancreatic cancer or detecting early-stage pancreatic cancer.

Language：English  

Language：English  

Pancreatic juice is a promising type of diagnostic sample for pancreatic cancer, and members of the mucin (MUC) family are diagnostic candidates. To evaluate the utility of MUC family members as diagnostic markers, we measured MUC mRNA expression in pancreatic tissues and pancreatic juice obtained from patients with different pancreatic diseases as well as in pancreatic cancer cell lines by real-time PCR. Furthermore, to support the possibility of early diagnosis by quantification of MUC1 and MUC5AC, immunohistochemistry and microdissection-based quantitative analysis of mRNA were carried out. There was no significant correlation between MUC1 and MUC5AC expression in cell lines. When beta-actin was used as a reference gene, median MUC1 and MUC5AC mRNA expression levels were remarkably greater in tumoral tissues than in non-tumoral tissues, but median MUC4 and MUC6 mRNA expression levels were not. Receiver operating characteristic curve analysis showed that quantitative analysis of MUC1 and MUC5AC mRNA in pancreatic juice is better diagnostic modality than that of MUC4 and MUC6 mRNA. Immunohistochemistry showed that MUC1 and MUC5AC were highly expressed in invasive ductal carcinomas (IDC) and moderately expressed in high-grade pancreatic intraepithelial neoplasia (PanIN); no staining was observed in normal ducts. Analysis of cells isolated by microdissection showed stepwise upregulation of MUC1 and MUC5AC in the development of high-grade PanIN to IDC. Our results suggest that MUC1 and MUC5AC are upregulated stepwise in pancreatic carcinogenesis and that quantitative assessment of MUC1 and MUC5AC mRNA in pancreatic juice has high potential for preoperative diagnosis of pancreatic cancer. Copyright 2005 Wiley-Liss, Inc.

Language：English  

Although telomerase activity is a promising diagnostic marker, clinical introduction of this marker for cancer diagnosis is still problematic due to the lack of means of evaluating sample quality. Human telomerase reverse transcriptase (hTERT), one of the subunits of telomerase, is also a promising diagnostic marker. In the present study, we did large-scale analysis of 88 pancreatic juice samples to determine the feasibility of quantitative analysis of hTERT mRNA for diagnosis of pancreatic cancer. We found significant differences in hTERT expression among carcinoma-derived, intraductal papillary mucinous neoplasm (IPMN)-derived, and chronic pancreatitis-derived juice samples. Results showed that quantitative analyses of hTERT mRNAs are more useful in discriminating carcinoma from IPMN than from chronic pancreatitis. When the specificity was set at 100%, the sensitivity for differentiation between carcinoma and IPMN was 43.5%, whereas the sensitivity of cytologic examination was 22.0%. There were significant differences in hTERT expression among carcinoma cells, IPMN cells, and normal ductal cells isolated from pancreatic tissues by microdissection. Lymphocytes and hyperplastic epithelial cells isolated from tissues with the histologic appearance of pancreatitis showed various expression levels of hTERT. Our results suggest that quantitative analysis of hTERT mRNA in pancreatic juice is advantageous over cytologic analysis for differentiation between carcinoma and IPMN but probably not for differentiation between carcinoma and chronic pancreatitis.

Language：English  

All-trans retinoic acid (ATRA) inhibits proliferation of cancer. However, the effects of ATRA on scattering and invasion of pancreatic cancer cells remain unknown. Also, the effects of ATRA on c-Met expression in pancreatic cancer have never been addressed so far. The effects of ATRA on a pancreatic cancer cell line, Capan-1, were determined by proliferation assay, scattering assay and invasion assay. In addition, the expression of c-Met in pancreatic cancer cell lines treated with ATRA was investigated by real-time PCR and western blotting. The growth-inhibitory effect of ATRA was found when the cells were cultured with 5 microM ATRA for 3 days. In cell scattering assay, ATRA-treated pancreatic cancer cells were found to spread out from their colonies. In invasion assay, cells treated with ATRA invaded the matrigel more than vehicle-treated cells. The expression of c-Met was up-regulated both in the mRNA and protein levels after the treatment of ATRA. The highest expression was found at 48 h after the treatment. ATRA induced scattering and invasion of pancreatic cancer cells, although it inhibited proliferation of those cells. In addition, ATRA also increased the protein level of c-Met. These findings may indicate that the use of retinoic acid as an anti-cancer therapeutic drug needs some additional treatments to control cell invasion or scattering.

Language：English  

Recent microarray analyses showed that the S100 family contains members that are candidate diagnostic markers or therapeutic targets. In the present study, to evaluate the involvement of S100A6 in pancreatic cancer and its clinical usefulness for diagnosis, we examined S100A6 mRNA expression in pancreatic tissues and pancreatic juice from patients with different pancreatic diseases. To investigate the role of S100A6 in carcinogenesis of pancreatic cancer and the potential of S100A6 as a diagnostic marker for early detection of pancreatic cancer, we did immunohistochemistry and microdissection-based mRNA analysis of pancreatic normal ducts, pancreatic intraepithelial neoplasias, and invasive ductal carcinomas. We also used in vitro experiments and microarray analysis with RNA interference to evaluate the functional role of S100A6 and its potential as a therapeutic target for pancreatic cancer. S100A6 mRNA levels were significantly higher in carcinoma specimens than in nonneoplastic tissues. In pancreatic juice, there was a significant difference in S100A6 expression between patients with carcinoma and those with nonneoplastic disease. Receiver operating characteristic curves revealed that S100A6 might be a useful marker for diagnosis of pancreatic cancer. Immunohistochemistry and microdissection-based analysis showed differential expression of S100A6 among normal ducts, pancreatic intraepithelial neoplasias, and invasive ductal carcinomas. In vitro data showed that inhibition of S100A6 decreased proliferation and invasiveness of cancer cells, and these findings were supported by microarray data. Our present results suggest that quantitation of S100A6 mRNA is a promising tool for diagnosis of pancreatic cancer, and that S100A6 may be a promising therapeutic target for pancreatic cancer.

Language：English  

NK4, a 4-kringle fragment of hepatocyte growth factor (HGF), is an HGF antagonist that also acts as an angiogenesis inhibitor. NK4 strongly inhibits the infiltration, metastasis, and tumor growth of pancreatic cancer. The aim of our study was to evaluate the antitumor effect of adenovirus-mediated NK4 gene transfer to the liver on hepatic metastasis of pancreatic cancer in vivo. We constructed recombinant adenoviral NK4 (Ad-NK4), which encodes a secreted form of human NK4. Intrasplenic injection of Ad-NK4 induced high and relatively maintained expression of NK4 protein in the liver and suppressed the number and growth of metastatic foci in the liver in a nude mouse model. Microscopically, central necrosis was found even in small metastatic foci in Ad-NK4 treated mice. Immunohistochemical analysis of metastatic tumors showed a remarkable decrease in microvessel density and an increase in the number of apoptotic tumor cells after treatment with Ad-NK4. These results indicate that intraportal injection of Ad-NK4 may be a useful therapeutic modality for the clinical control of hepatic metastasis in pancreatic cancer. Copyright (c) 2005 Wiley-Liss, Inc.

Language：English  

Measurement of telomerase activity is a promising diagnostic tool for pancreatic cancer. Detection of mRNA for human telomerase reverse transcriptase (hTERT), a catalytic subunit of telomerase, is also a diagnostic candidate. In the present study, we developed a telomeric repeat amplification protocol assay with real-time PCR and a protocol for quantification of hTERT mRNA with real-time PCR. To evaluate the feasibility of these methods for diagnosis of pancreatic cancer, we measured telomerase activity and hTERT expression in pancreatic cancer cell lines, pancreatic tissues, and pancreatic juice samples from patients with different pancreatic diseases. There were significant correlations between telomerase activity and hTERT expression in cell lines, tissues, and juice samples. The levels of telomerase activity and hTERT expression were significantly higher in tumoral tissues than in nontumoral tissues. In pancreatic juice specimens, some carcinoma samples showed remarkably high expression of hTERT. However, there were no significant differences in hTERT expression between patients with carcinoma and those with benign diseases, although significant differences in telomerase activity were observed. Our present results suggest that the combined assessment of hTERT and telomerase activities in pancreatic juice provides a potent diagnostic method for pancreatic cancer.

Language：English  

Abstract

BACKGROUND:

Early detection of pancreatic carcinoma is difficult even with current diagnostic tools. Novel biomarkers and detection techniques are urgently needed. Telomerase activity is a promising diagnostic marker. However, the conventional telomeric repeat amplification protocol (TRAP) assay is not suitable for clinical application because of its complexity, time-consuming nature, and the effects of polymerase chain reaction (PCR) inhibitors in samples leading to difficulties in quantification.

METHODS:

The authors used a hybridization protection assay in combination with TRAP (TRAP/HPA) to investigate the effects of PCR inhibitors in pancreatic juice on quantification of telomerase activity. They analyzed 117 consecutive samples of pancreatic juice to determine the feasibility of TRAP/HPA for diagnosis of pancreatic carcinoma.

RESULTS:

The authors found that TRAP/HPA was 1000-fold more sensitive than the conventional TRAP assay, and that the effects of PCR inhibitors could be avoided by diluting samples. In a large analysis of pancreatic juice samples with TRAP/HPA, 17 samples were excluded from the final analysis because of insufficient follow-up periods or inadequate treatment of the samples. Relative telomerase activity (RTA) in samples from patients with pancreatic carcinoma was significantly higher in comparison to samples from patients with pancreatitis and 13 (61.9%) of 21 samples from patients with pancreatic carcinoma showed high RTA (> 4 U). Meanwhile, high RTAs were observed in 4 of 35 (11.4%) samples from patients with intraductal papillary mucinous tumor and in 1 of 40 samples (2.5%) fom patients without malignant disease.

CONCLUSIONS:

TRAP/HPA accurately evaluated weak telomerase activity in pancreatic juice samples without the problem due to PCR inhibitors. This large analysis of nonselected pancreatic juice samples suggested that TRAP/HPA is a promising approach for the diagnosis of pancreatic carcinoma.

Language：English  

Radiotherapy represents a major treatment option for patients with pancreatic cancer, but recent evidence suggests that radiation can promote invasion and metastasis of cancer cells. Interactions between cancer cells and surrounding stromal cells may play an important role in aggressive tumor progression. In the present study, we investigated the invasive phenotype of pancreatic cancer cells in response to coculture with irradiated fibroblasts. Using in vitro invasion assay, we demonstrated that coculture with nonirradiated fibroblasts significantly increased the invasive ability of pancreatic cancer cells and, surprisingly, the increased invasiveness was further accelerated when they were cocultured with irradiated fibroblasts. The hepatocyte growth factor (HGF) secretion from fibroblasts remained unchanged after irradiation, whereas exposure of pancreatic cancer cells to supernatant from irradiated fibroblasts resulted in increased phosphorylation o
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c-Met (HGF receptor) and mitogen-activated protein kinase activity, possibly or partially via increased expression of c-Met. We also demonstrated that scattering of pancreatic cancer cells was accelerated by the supernatant from irradiated fibroblasts. The enhanced invasiveness of pancreatic cancer cells induced by coculture with irradiated fibroblasts was completely blocked by NK4, a specific antagonist of HGF. These data suggest that invasive potential of certain pancreatic cancer cells is enhanced by soluble mediator(s) released from irradiated fibroblasts possibly through up-regulation of c-Met expression/phosphorylation and mitogen-activated protein kinase activity in pancreatic cancer cells. Our present findings further support the potential use of NK4 during radiotherapy for patients with pancreatic cancer.

Language：English  

The intensive stromal reaction is one of characteristics of pancreatic exocrine carcinoma. The mutual interaction between pancreatic cancer cells and orthotopic tumor-derived fibroblasts have not been clarified yet. In this study, we sought to elucidate the mechanism underlying the tumor-stromal interaction with an in vitro coculture experimental system. Considerable strong c-Met expression was detected in seven out ten lines of human pancreatic carcinoma cells, as determined by Western blotting. For hepatocyte growth factor (HGF)-production, however, none or only trace amounts of HGF could be detected in those ten cell lines. Of the two lots of tumor-derived fibroblasts obtained from two pancreatic cancer patients, the fibroblasts capable to produce HGF could initiate an apparent invasion-stimulating response in strong c-Met-expressed Suit-2 and Panc-1 cells but not in faint expressed Mia PaCa-2 and BxPC-3 cells. A specialized HGF antagonist, NK4 would effectively inhibit the fibroblast-mediated invasive growth, thus proving the key role of the paracrine-fashioned HGF/c-Met pathway in the tumor-stromal interaction. On the other hand, the regulative action of cancer cells on HGF expression of fibroblasts was also investigated using direct or indirect coculture systems. For the fibroblasts that originally did not produce HGF, cancer cells failed to show any HGF-inductive effect. For the HGF-producing fibroblasts, despite of somewhat upregulation or downregulation in fibroblast HGF expression, the feedback regulation by studied pancreatic cancer cells in both coculture modes were relatively limited. This in vitro study sketched out the interaction between cancerous and stromal compartments with an emphasis on HGF/c-Met signal pathway, thus possibly helping to unveil the more complicated mutual modulation in vivo between pancreatic cancer and host mesenchymal tissues.

Language：English  

OBJECTIVE: In this retrospective review, we evaluated the advantages and disadvantages of LADG for patients of heavier weight with early gastric cancer. SUMMARY BACKGROUND DATA: LADG has been used to treat early gastric cancer. We and others have reported less operative blood loss, less pain, early recovery of bowel activity, early restart of oral intake, and a shorter hospital stay with LADG compared with a conventional open method. There is, however, little information on the advantages of LADG for obese patients with early gastric cancer. METHODS: Between January 1996 and March 2002, 76 patients with preoperatively diagnosed early gastric carcinoma underwent LADG in our department. We classified these patients into 2 groups on the basis of body mass index (BMI). Nineteen patients had a high-BMI (>/= 24.2 kg/m2), and 57 patients had a normal-BMI (<24.2 kg/m2). We collected data by retrospectively reviewing the medical charts. RESULTS: Extension of the minilaparotomic incision or conversion to laparotomy was needed in 6 (32%) of the 19 patients in the high-BMI group, whereas only 3 (5%) of 57 patients in the normal-BMI group required either. In the high-BMI group, Roux-en-Y anastomosis rather than Billroth I anastomosis was adopted more often than in the normal-BMI group, due to the difficulty of the reconstruction (58% versus 4%, P = 0.001). Significantly longer operative time (370 +/- 61 minutes versus 317 +/- 58 minutes, P = 0.015) and prolonged recovery of bowel activity (3.5 +/- 1.0 days versus 2.6 +/- 1.0 days, P = 0.007) were observed in the patients in the high-BMI group. CONCLUSIONS: In the current study, LADG in patients of heavier weight was accompanied by more technical difficulties, and the disadvantages of longer operative time and delayed recovery of bowel activity was observed in patients of heavier weight. Heavier weight appears to be an ominous factor in the successful completion of LADG and should be considered in the decision to use LADG. There are still benefits of a decreased incidence of serious wound and hernia complications in successful cases.

Responsible for pages：立体映像技術－空間表現メディアの最新動向－,140-143   Language：Japanese  

Responsible for pages：がん転移研究の実験手法,241-247   Language：Japanese  

Responsible for pages：感覚、感情とロボット,361-371   Language：Japanese  

Event date： 2024.4

Venue：名古屋市   Country：Japan  

Event date： 2024.4

Venue：常滑市   Country：Japan  

【背景】
食道癌を含む固形癌では腫瘍浸潤リンパ球（TIL）が良好な予後や免疫療法の効果と相関していることが多く報告されている。さらに、腫瘍微小環境（TME）におけるTILの局在と密度によっていくつかの分類法が存在し、より正確な予後因子、治療効果予測マーカーとなっている。しかし、これらのTMEによる分類の分子生物学的な意義は十分には理解されておらず、その臨床的な意義につながるメカニズムに関しては不明である。
【目的】
食道癌のTMEサブタイプにおける腫瘍免疫を中心とした機能的な不均一性を解明する
【対象と方法】
2019年5月～2023年5月までにsingle-cell RNA sequencing を行った切除食道扁平上皮癌27例に対して免疫組織化学染色を行った。腫瘍部におけるCD8陽性細胞の局在と密度を定量的に評価し、”hot”, “excluded”, “cold”の3つに分類した。それぞれ3例ずつ、計9例を対象としてsingle-cell RNA解析を行った。
【結果】
全9例の統合解析を行い、Quality Checkおよびdoublet cellの除去後、39790細胞が得られた（hot:13840 cells, excluded:11168 cells, cold:14782cells）。8つのクラスター（T cell, Myeloid cell, B cell, Plasma cell, Fibroblast, Endothelial cell, Epithelial cell, MAST cell）に分類され、”excluded”ではFibroblastの割合が多く、T cellの割合が少なかった。続いてCD8陽性T細胞を抽出し、4155細胞が得られた。全細胞に占めるCD8陽性T細胞の割合は”hot”, “cold”, “excluded”の順に多かった。TME分類ごとに遺伝子発現解析を行った。”hot”ではIFNGが最も高発現であり、細胞障害性を示す”cytotoxic score”は最も低く、疲弊化を示す”exhaustion score”は最も高かった。
【結語】
hot tumorのCD8陽性T 細胞はIFNGが高発現であることに伴い、疲弊化が進んでおり、細胞障害性が低下していることが示唆される。

Event date： 2024.4

Venue：名古屋市   Country：Japan  

Event date： 2024.4

Venue：名古屋市   Country：Japan  

Event date： 2024.4

Venue：名古屋市   Country：Japan  

【はじめに】
膵癌は予後不良な疾患であり、その一因としてT細胞浸潤が乏しい抑制性免疫微小環境、いわゆるcold tumorであることが知られている。他癌腫ではcold tumorをT細胞浸潤のあるhot tumorへと誘導することで免疫チェックポイント阻害剤が奏功したという報告もあり、免疫細胞浸潤機序の解明が待たれている。膵癌組織において、癌細胞のどのような因子が免疫細胞浸潤を促進するのかについては、いまだ明らかになっておらず、本研究ではその解明を目的とした。
【方法】
膵癌自然発癌マウス由来の膵癌細胞株を複数樹立し、それぞれ免疫のあるマウスの膵臓に同所移植した。3週後の膵癌組織を免疫染色し、CD8 T細胞浸潤の程度からhot/cold TIME(腫瘍免疫微小環境)に分類した。各TIMEを誘導する膵癌細胞株を3種類ずつ選出し、RNAシークエンスを行い、hot TIMEを誘導する膵癌細胞の発現変動遺伝子を同定した。さらにヒト膵癌のpublic data (Peng et al, 2019)をシングルセルRNA解析し、発現変動遺伝子のうちよりCD8 T細胞浸潤に関与する可能性のある遺伝子を同定した。
【結果】
マウス移植実験により、細胞数や移植後の観察期間に関わらず、常にhot TIME(腫瘍内にCD8 T細胞浸潤が多い)もしくはcold TIME(T細胞浸潤が乏しいもしくは辺縁にのみ集族する)を呈する細胞株をそれぞれ3種類ずつを同定した。これらの癌細胞株のRNAシークエンスにより、hot TIMEとなる癌細胞株群で22の発現変動遺伝子を抽出した。シングルセルRNA解析を行い、CD8 T細胞の細胞数などから抑制性免疫微小環境を呈する患者群では、これらの遺伝子群の発現が低いことを確認した。また発現変動遺伝子のうちCSF1の発現が高い患者ではCD8T細胞発現細胞数が有意に低いことを明らかにした。
【まとめ】
異なる遺伝子発現の膵癌細胞株の移植では異なる免疫微小環境を呈することを明らかにした。さらにhot TIMEの誘導に関与する可能性のある遺伝子を同定した。

Event date： 2024.3

Venue：北九州市   Country：Japan  

Event date： 2024.3

Venue：大分市   Country：Japan  

Event date： 2024.3

Venue：大分市   Country：Japan  

Event date： 2024.3

Venue：大分市   Country：Japan  

Event date： 2024.2 - 2024.3

Venue：京都市   Country：Japan  

Event date： 2024.2 - 2024.3

Venue：京都市   Country：Japan  

Event date： 2024.2 - 2024.3

Venue：京都市   Country：Japan  

【背景・目的】近年、免疫療法は胃癌治療に革新をもたらしたが、Nivolumabの奏効率は約40%であり、半数以上で奏功しない症例が存在する。Nivolumab不応症例における腫瘍微小環境については未だ不明な点が多く、本研究では、Nivolumab不応症例の腫瘍微小環境内に存在するCD8陽性T細胞と骨髄由来抑制細胞(MDSC)の機能関連遺伝子の発現を明らかにすることを目的とした。
【方法】当科で胃切除術を施行した胃癌患者から正常粘膜部3サンプル、未治療腫瘍部3サンプル、化学療法後腫瘍部(PR) 2サンプル、化学療法+Nivolumab治療後(PD)腫瘍部2サンプルを採取し、single-cell RNA sequencingを行い、各群で機能関連遺伝子の発現を比較検討した。
【結果】CD8陽性T細胞は遺伝子発現の特徴から7つのクラスターに分類された。腫瘍抗原特異的細胞障害性機能が高いと報告されている、Progenitor exhausted CD8陽性T細胞のクラスターでは化学療法+Nivolumab治療後(PD)腫瘍部の細胞障害性機能関連遺伝子発現が他の群と比較して有意に高値であった。さらにProgenitor exhausted CD8陽性T細胞のクラスターの割合は未治療腫瘍部・化学療法後腫瘍部よりも多く、正常粘膜部とほぼ同じであった。次に免疫抑制細胞であるMDSCに着目して検討したところ、単球系MDSCで化学療法+Nivolumab治療後(PD)腫瘍部の免疫抑制性機能関連遺伝子発現が他の群と比較して有意に高値であった。一方、多形核系MDSCでは化学療法+Nivolumab治療後(PD)腫瘍部の免疫抑制性機能関連遺伝子発現は未治療腫瘍部・化学療法後腫瘍部と比較して有意な差は認めなかった。
【結論】単球系MDSCにおける免疫抑制機能の上昇がNivolumab不応の一因である可能性が示唆された。

Event date： 2024.2 - 2024.3

Venue：京都市   Country：Japan  

Event date： 2023.12

Venue：北九州市   Country：Japan  

大腸手術におけるSSIはその他の消化管手術に比較してリスクが高く、SSIの発生が癌の予後に影響するという報告もあり、SSIの予防は大腸外科医にとって非常に重要な命題である。特に直腸手術では縫合不全などの臓器/体腔SSIも含めてリスクが高い。ロボット支援下手術は多関節機能・手振れ防止機能のロボットアームと安定した視野を生む3Dカメラにより、特に骨盤のような狭い空間での操作性・安定性が可能でより精緻な手術が可能になる。ロボット支援下手術における当科でのSSI予防のためのケアバンドルについて概説する。周術期支援センターにて術前禁煙指導、栄養指導や運動指導、糖尿病患者には血糖コントロール、歯科と連携した口腔ケアを行う。低侵襲手術を励行し、腸管前処置及び術前化学的腸管処置、などを術前に施行する。手術室では直前に最低限の除毛を行い、予防的抗菌薬を執刀前に投与し、皮膚消毒を施行する。術中は手袋交換(３時間ごと、閉創前)、ウォーマーを使用して十分な保温を行い、ダブルリング創縁保護、モノフィラメント抗菌縫合糸を使用する。キ)タタ旅估睚Ⅴ膸楾垰・砲歪牡紐・鋠・屬浪椎修文造蠱擦・靴謄・璽爾箋朧悊覆匹魘郢箸靴督夏睛栃・任留・・鮑把禪造砲垢襦２椎修文造螢疋譟璽鵑倭淨・擦此∧珍倭阿砲牢鏗８魎垢掘∧珍六・砲倭論・・擇喊身號ス腓鮖楾圓垢襦・儻紊魯疋譟璽鵑篩鷲瑤亙頂織疋譽奪轡鵐阿鮖楾圈▲疋譟璽鵑浪椎修文造蠢甦釮鉾患遒垢襦SSIのサーベイランスを行って当科におけるSSI発生状況を監視する、などのSSI対策をバンドルとして施行している。当科にて2018年10月から2023年8月に施行したロボット支援下大腸手術症例135例におけるSSIリスク因子の検討を行い、SSI予防に有効な対策を明らかにする。

Event date： 2023.12

Venue：北九州市   Country：Japan  

Event date： 2023.12

Venue：北九州市   Country：Japan  

大腸手術におけるSSIはその他の消化管手術に比較してリスクが高く、SSIの発生が癌の予後に影響するという報告もあり、SSIの予防は大腸外科医にとって非常に重要な命題である。特に直腸手術では縫合不全などの臓器/体腔SSIも含めてリスクが高い。ロボット支援下手術は多関節機能・手振れ防止機能のロボットアームと安定した視野を生む3Dカメラにより、特に骨盤のような狭い空間での操作性・安定性が可能でより精緻な手術が可能になる。ロボット支援下手術における当科でのSSI予防のためのケアバンドルについて概説する。周術期支援センターにて術前禁煙指導、栄養指導や運動指導、糖尿病患者には血糖コントロール、歯科と連携した口腔ケアを行う。低侵襲手術を励行し、腸管前処置及び術前化学的腸管処置、などを術前に施行する。手術室では直前に最低限の除毛を行い、予防的抗菌薬を執刀前に投与し、皮膚消毒を施行する。術中は手袋交換(３時間ごと、閉創前)、ウォーマーを使用して十分な保温を行い、ダブルリング創縁保護、モノフィラメント抗菌縫合糸を使用する。キ)タタ旅估睚Ⅴ膸楾垰・砲歪牡紐・鋠・屬浪椎修文造蠱擦・靴謄・璽爾箋朧悊覆匹魘郢箸靴督夏睛栃・任留・・鮑把禪造砲垢襦２椎修文造螢疋譟璽鵑倭淨・擦此∧珍倭阿砲牢鏗８魎垢掘∧珍六・砲倭論・・擇喊身號ス腓鮖楾圓垢襦・儻紊魯疋譟璽鵑篩鷲瑤亙頂織疋譽奪轡鵐阿鮖楾圈▲疋譟璽鵑浪椎修文造蠢甦釮鉾患遒垢襦SSIのサーベイランスを行って当科におけるSSI発生状況を監視する、などのSSI対策をバンドルとして施行している。当科にて2018年10月から2023年8月に施行したロボット支援下大腸手術症例135例におけるSSIリスク因子の検討を行い、SSI予防に有効な対策を明らかにする。

Event date： 2023.12

Venue：横浜市   Country：Japan  

Event date： 2023.12

Venue：横浜市   Country：Japan  

【背景・目的】
直腸癌手術後の縫合不全(AL)は頻度の高い合併症の一つである。当科におけるALを0にするための取り組みについて検討し、その有用性を明らかにすることを目的とした。
【当科の周術期管理】
前日より絶食にし、成分栄養剤を内服する。2020年4月より機械的腸管処置に化学的腸管処置を追加し、ICG 蛍光造影法により術中腸管血流評価を行っている。経肛門ドレーン留置や経肛門的に吻合部補強を行い、術前治療症例や吻合部が低い症例は一時的回腸人工肛門造設術を行う。近年はロボット支援下手術で全例行っている。
【対象・方法】
対象は、2016年1月から2023年3月にDST/SST吻合を施行した直腸癌115例。化学的腸管処置、ICG蛍光造影法を開始した2020年4月以降の52例(N群)と、以前の63例(O群) で比較検討した。
【結果】
年齢、性別、BMI、ASA-PS、腫瘍占拠部位、術前CAR、PNI、手術時間、出血量、腫瘍最大径、最終Stage、術後在院日数において両群間で差を認めなかった。ロボット支援下手術はN群45例、O群9例であった(p<0.0001)。ALはN群2例、O群9例で、Clavien-dindo分類Grade3a以上はN群では認めず、O群14%(9/63)より有意に低かった(p=0.0039)。
【結語】
現在当科で施行している直腸癌手術の縫合不全への対策は有用であると思われた。

Event date： 2023.12

Venue：横浜市   Country：Japan  

Event date： 2023.12

Venue：横浜市   Country：Japan  

＜背景＞
当科では現在までに95例のMIPD＜LPD（28例）、RPD(67例)＞を行っている、MIPDに必要な解剖を紹介、当院で行っている膵癌に対する術式に関して紹介する。
＜方法>
門脈に接する膵癌に対してはmesenteric approachを行うことがある。本操作では中結腸動脈をメルクマールとしてSMAを同定する。SMA周囲を根部に向かって剥離を行い、その後treitz靭帯を切離する。Treitz靭帯は半数以上が十二指腸背側にも付着しており前方のみならず、背側からも確実に切離する。可能であればSMA,SMVの間でIPDAもしくは1st JAを切離するがこの時注意すべきは1st JVである、1stJVは殆どがSMA背側を走行しているため、出血に注意する。膵を離断後は、Hanging maneuverを利用してSMV右側から腹側を展開して剥離を行う。
＜結果＞
95例のMIPD中、膵癌に対するRPD は現在までに27例 施行した（stage 0 10例、IA 6例、IIA 5例、IIB 6例）。手術時間中央値および出血量中央値はそれぞれ733分（437分－863分）、235g （0－1050g）であった。長期成績に関しては、観察期間中央値21ヶ月（1－59ヶ月）であり、1例に再発を認めたが、そのほかの症例に関しては無再発生存中である。

Event date： 2023.12

Venue：横浜市   Country：Japan  

Event date： 2023.12

Venue：横浜市   Country：Japan  

Event date： 2023.12

Venue：横浜市   Country：Japan  

大腸全摘は潰瘍性大腸炎(UC)及び家族性大腸腺腫症(FAP)の標準術式である。手術行程が多く、手術時間も長時間となり低侵襲性から腹腔鏡手術が導入されつつある。経肛門的内視鏡手術は経肛門/会陰的に逆行性に直腸固有筋膜レベルでの剥離を行うアプローチ方法で、腹腔操作と同時に行うことにより手術時間短縮だけでなく、双方向からの確認が可能となり、安全な手術の一助となる。FAPでは炎症もなく授動操作は比較的容易だが、UCでは随伴する炎症により剥離に難渋することもある。なお当科ではUCとFAPで手術手順は変えていない。2020年より大腸全摘術に経肛門内視鏡手術併用を開始した。
現在までに当科では経肛門的内視鏡手術併用大腸全摘術をUC７症例及びFAP３症例に施行した。年齢はFAP23.7歳、UC55.9歳と明らかな差があった(P=0.012)。性別はFAP男性1、女性2、UC男性5、女性2とUCで男性が多い傾向にあった。FAPは全例IAAを施行、UCでは3例がIACA、1例がIAA、2例が全摘、回腸人工肛門造設であった。手術術式の違いは罹患範囲や患者年齢が関与していると考えられる。手術時間は中央値でFAP423分、UC410分であり、出血量はそれぞれ168g、170gであった(p=0.643、p=0.766)。1例UCで出血による開腹移行を経験した。術後Clavien-Dindo Grade 2の腸閉塞と会陰部SSI、腹腔内膿瘍をそれぞれ1例に認め、全てUC症例であった。UCとFAPでは切除範囲、再建の有無などに違いがあり、一概には言えないが、炎症の影響により、UCの方が術後合併症が多く、手術難易度は高い。

Event date： 2023.12

Venue：横浜市   Country：Japan  

Event date： 2023.12

Venue：横浜市   Country：Japan  

Event date： 2023.12

Venue：横浜市   Country：Japan  

Event date： 2023.12

Venue：横浜市   Country：Japan  

Event date： 2023.11

Venue：岡山市   Country：Japan  

大腸癌治療ガイドラインではStage IV大腸癌は遠隔転移/原発巣が切除可能な場合、原発巣の根治切除とともに遠隔転移の切除が強く推奨されている。しかし、Conversion surgeryについては、遠隔転移臓器の切除前後の化学療法施行の是非などに一定の見解はなく、再発巣切除のタイミングの判断や再発や予後の予測は難しい。
今回、Stage IV大腸癌に対してR0切除術を施行した85症例において、遠隔転移切除前後の化学療法施行の予後への影響を検討した。遠隔転移の同時切除が39症例、異時切除が46症例であった。初発時の遠隔転移臓器は肝47例、肺11例、播種20例、リンパ節14例、卵巣2例で同一臓器の多発症例は肝で22例、肺2例、2臓器以上にわたって転移巣が多発するものは10例であった。異時切除症例では肝34例、肺11例、播種4例、リンパ節1例であり、薬物治療施行のタイミングは原発巣切除前17例、転移巣切除前27例、R0達成後60例であった。異時性に転移巣を切除する症例では(1)転移巣切除前薬物療法を行わない16症例(肝7、肺8、播種1)、(2)原発巣切除を先行し、Conversionが出来るタイミングまで行う分子標的薬を併用した14症例(肝12、播種2)、(3)原発巣切除を先行し、切除可能な遠隔転移を切除する前に施行した16症例(肝15、肺3、播種1キ)タ氈」螢鵐兩1)、がある。(2)については今回解析した症例以外にConversionを目指していたが転移巣制御できず手術不能となった症例も10例認めた。2臓器以上にわたって転移巣が多発するものは(3)に多く、R0切除が達成できた2臓器以上の遠隔転移症例の遠隔転移は切除可能で微小転移制御のために遠隔転移切除前に化学療法を施行している症例が多かった。予後について解析を行ったところ、(1)と(3)では薬物療法を行わない方がCSS不良の傾向があり(P=0.109)、(2)と(3)ではConversion症例の方がわずかにCSS不良であったものの有意差は認めなかった(P=0.302)。Conversion症例でもR0切除が達成できれば切除可能症例と予後に明らかな差は認めず、Conversion surgeryについては積極的に検討すべきである。

Event date： 2023.11

Venue：岡山市   Country：Japan  

Event date： 2023.11

Venue：岡山市   Country：Japan  

Event date： 2023.11

Venue：San Diego  

Background: Pancreatic cancer is known to accumulate many mutations, such as KRAS and TP53, in carcinogenesis, but early detection using liquid biopsy is still difficult. We aimed to analyze genetic alternations that accumulate from the precursor lesion to tumorigenesis and cancer progression, including liquid biopsy, and to find key genetic mutations at each stage. We also attempted to investigate effect of those key mutations on tumor microenvironment.
Methods: Peripheral blood (PB) was collected once a week from KPC mice older than 7-weeks of age until sacrifice and genomic DNA was extracted after hemolysis. For sorting of circulating tumor cells (CTCs) in PB and hemorrhagic ascites of sacrificed KPC mice, anti-mouse CD45 MicroBeads was used. Genomic DNA was extracted from CTCs, and FFPE samples of primary pancreas and liver metastases. We performed exome sequencing with NovaSeq.
Results: KPC mice were divided into three groups (no primary, primary/no metastasis, and primary/metastasis), and histological images were observed at each stage of cancer progression. When genetic mutations were compared between chronological CTCs of PB and FFPE samples of primary pancreas, we found two mutations, one of which was related to MHC class I. On the other hand, no MHC I mutation were found in the no primary group. When the effect of MHC I mutation on immune cells was evaluated, primary pancreatic tissue showed CD8+ T cell cluster in the regional lymph node compared with no primary group. Two mutations, including MHC I mutation, that appeared at early tumorigenesis were dominant in PB and hemorrhagic ascites, while were not detected in liver metastasis.
Conclusions: The present data suggest that MHC I mutation which circulate in blood from early carcinogenesis can be an indicator of early detection and that the immune response may eliminate cancer cells with MHC I mutation in liver metastases.

Event date： 2023.11

Venue：San Diego  

Event date： 2023.11

Venue：San Diego  

Event date： 2023.11

Venue：San Diego  

Event date： 2023.11

Venue：熊本市   Country：Japan  

Event date： 2023.11

Venue：熊本市   Country：Japan  

Event date： 2023.11

Venue：神戸市   Country：Japan  

Event date： 2023.11

Venue：神戸市   Country：Japan  

Event date： 2023.11

Venue：神戸市   Country：Japan  

【背景】近年、腫瘍免疫抑制性微小環境における多形核系骨髄由来抑制細胞 (PMN-MDSC)が着目されており、担癌患者の末梢血、リンパ組織、転移組織におけるPMN-MDSCの報告が増加している。しかし、胃癌浸潤PMN-MDSCの役割や機能については未だ不明な点が多い。
【目的】胃癌浸潤PMN-MDSCの機能や臨床的予後との相関を明らかにする。
【対象・方法】
(1) 当科で胃切除術を施行した症例から採取した胃癌組織12例と隣接正常組織10例のsingle-cell RNA sequencing (scRNA-seq)を行い、PMN-MDSCにおける免疫抑制機能関連遺伝子を比較検討した。
(2) 胃癌組織12例と隣接正常組織10例のPMN-MDSCを多重免疫染色で評価した。
(3) 既に報告されている胃癌27例のscRNA-seqで細胞障害性CD8陽性T細胞とPMN-MDSCを含めた複数の免疫抑制細胞との相関を検討した。
(4) 細胞間相互作用解析でPMN-MDSCと複数の免疫細胞との相互作用を評価した。
(5) 胃切除術を施行した胃癌患者52例における胃癌浸潤PMN-MDSCの細胞数と臨床的予後の相関を評価した。
【結果】
(1) PMN-MDSCにおける免疫抑制機能関連遺伝子の発現は正常隣接組織と比較して胃癌組織で有意に高値であった (P <0.0001)。
(2) 胃癌組織では隣接正常組織よりもPMN-MDSCが有意に多かった (P = 0.0169)。
(3) 細胞障害性CD8陽性T細胞数が少ない症例で最も多い免疫抑制細胞はPMN-MDSCであった。
(4) PMN-MDSCにおけるIL1Bと細胞障害性CD8陽性T細胞におけるIFNGは細胞間で相互作用がある可能性が示唆された。
(5) 胃癌浸潤PMN-MDSCが多い群26例と少ない群26例に分類した。PMN-MDSCが多い群はPMN-MDSCが少ない群と比較して無再発期間と術後生存期間が有意に短かった (P = 0.0003, P = 0.0132)。
【結論】胃癌浸潤PMN-MDSCは免疫抑制性微小環境の形成に重要である可能性が示された。

Event date： 2023.11

Venue：神戸市   Country：Japan  

【目的】近年、食道癌において免疫チェックポイント阻害薬(ICB)と化学療法の併用療法の有効性が示され化学療法に伴う腫瘍免疫微小環境の変化が注目されている。また腫瘍局所に存在する腫瘍浸潤Bリンパ球(TIL-B)は複数の癌腫の予後良好因子であることが報告されているが、腫瘍局所での機能的な役割に関しては不明である。今回TIL-Bの化学療法に伴う機能変動を評価するため食道癌検体を用いたシングルセル解析を行った。
【方法】食道扁平上皮癌10検体（化学療法施行群５検体、未施行群５検体）、正常粘膜部7検体の計17検体を用いてシングルセル解析を行った。また、食道扁平上皮癌166検体を対象に抗体産生細胞マーカーの抗IGKC抗体を用いた免疫組織化学染色(IHC)を行い、その臨床的意義を検討した。
【成績】17検体を対象にシングルセル解析を行い、81,246細胞を同定し24のクラスターに分類した。次にTIL-Bクラスターのみを抜き出し、12の詳細な分化サブタイプを同定した。さらに化学療法施行の有無に着目した検討ではTIL-B全体におけるCD40シグナル関連遺伝子の発現が化学療法施行群で増強していた(P＜0.0001)。ナイーブBリンパ球は化学療法施行群でBリンパ球活性化関連遺伝子の発現が増強し細胞数が減少していた(P=0.0064)。一方で抗体産生細胞は化学療法施行群において遊走能が低下しており(P＜0.0001)、腫瘍局所の細胞数が増加し(P=0.0134)、さらに抗体産生能も増強していた。IHCにてIGKC陽性細胞の出現頻度はRFS・OSの延長と有意に相関しており、多変量解析においても独立した予後良好因子であることが示された(RFS: HR: 0.23, P＜0.0001; OS: HR: 0.20, P=0.0002)。
【結論】本検討により化学療法に伴うTIL-Bの機能的不均一性の変動が明らかになり、消化器癌の腫瘍免疫微小環境のより詳細な理解につながるものと期待される。

Event date： 2023.11

Venue：神戸市   Country：Japan  

Event date： 2023.11

Venue：神戸市   Country：Japan  

【緒言】潰瘍性大腸炎(UC)は大腸粘膜にびらんや潰瘍ができる原因不明の大腸炎症性疾患である。発症は20代に多く、発症後10年以上の持続する炎症によりDysplasiaから腺癌の発生を見ることがあるが稀に神経内分泌腫瘍の発生を認める。
【方法】当科において2001年から2016年に手術を施行した潰瘍性大腸炎患者136例のうち神経内分泌腫瘍成分を伴う6例についてその臨床病理学的因子を検討した。
【結果】性別は男性2名、女性4名、平均年齢は47.1歳であった。罹患期間は全例10年を超えており、全例全大腸炎型であった。緊急手術をステロイド抵抗性の穿孔性腹膜炎、中毒性巨大結腸症のそれぞれ1例ずつに施行した。術前の薬物療法としてはステロイドは3例に使用されており、3例に免疫抑制剤使用歴があった。手術適応としては2例が緊急手術、2例が難治性もしくはステロイド依存性、残り2例は併存するUC関連大腸癌によるものであった。手術は全例大腸全摘で緊急手術の一例の開腹手術以外は腹腔鏡手術であった。術後病理組織診断では全例に活動性炎症を伴う活動期UCであり、2例はNET G1、2例は微小な内分泌細胞胞巣、1例は神経内分泌癌、1例は神経内分泌分化を伴う腺癌であった。全例背景粘膜にDysplasiaを認め、神経内分泌分化を伴う腫瘍の発生に潰瘍性大腸炎が関与している可能性が示唆された。
【結語】潰瘍性大腸炎において神経内分泌分化を伴う腫瘍の発生がまれにあり、それらを念頭においた診療が必要である。

Event date： 2023.11

Venue：神戸市   Country：Japan  

Event date： 2023.11

Venue：神戸市   Country：Japan  

[Background] Pancreatic ductal adenocarcinoma (PDAC) is a highly aggressive tumor with a complex microenvironment. Within this microenvironment, cancer-associated fibroblasts (CAFs) directly and indirectly interact with various immune cells, stromal cells, and cancer cells. CAFs can be classified into different subtypes, including myofibroblastic CAFs (myCAFs) and inflammatory CAFs (iCAFs). Transformed growth factor-beta 1 activated kinase-1 (TAK1) serves as a critical cellular hub for several cytokine-mediated signaling pathways that regulate inflammatory responses, such as NF-κB. However, the precise role of TAK1 in CAFs remains unclear. [Purpose]To explore TAK1 functions in CAFs in the PDAC microenvironment.[Materials and Methods] CAFs were primarily cultured from fresh resected pancreatic cancer specimens. Three-dimensional directly/indirectly co-culture models of PDAC tumors and CAFs were made to explore how tumor growth and mi
gration are influenced by TAK1+ CAFs. Then we checked the functions of the TAK1 in CAFs by cytokine array and western blotting (WB) using TAK1 inhibitor and siRNA of TAK1. Next, C57BL/6 mice were subcutaneously injected with organoids mixed with CAFs from KPC mice. [Results]Interference with TAK1 in CAFs inhibited tumor cell EMT and outgrowth in vitro. As well, TAK1 inhibitors decreased tumor growth and increased immune cell infiltration in the stroma in vivo.[Conclusions]TAK1 has a possibility to be a therapeutic target focusing on the CAFs which is a new direction for combined immunotherapy in PDAC.

Event date： 2023.11

Venue：神戸市   Country：Japan  

A 57-years-old female underwent operation for ovarian cancer seven years ago, and chemotherapy and radiation therapy were performed for the liver metastasis and recurrence around the round ligament. She was undergoing additional chemotherapy for the increasing dissemination in the splenic hilum, but she had epigastralgia and melena just before the chemotherapy. An upper gastrointestinal endoscopy showed duodenal ulcer scar without bleeding, then chemotherapy was started. However, she had abdominal pain three days after chemotherapy and enhanced CT scan revealed copious ascites with free air around the duodenum.
We performed operation for this patient on emergency. A hole of duodenal located in the bulbus, and it was a half circumferential, but the edge of duodenal wall had enough thickness to close, then simple closure using Gambee method with pedicled fat covering was performed. Naso-gastric tube was placed in the duodenum for decompression, and air leak test was negative. Moreover, we added Billroth-II type gastrojejunostomy in the case of duodenal stenosis after suturing. She was in good course, but a large amount of melena was occurred on POD6. Laboratory data showed elevated hepatobiliary pancreatic enzymes (AMY: 870 U/L AST: 90 U/L ALT: 49 U /L LDH: 253 U /L γ-GTP: 1656 U /L, Total Bilirubin 7.7U/L ), and decreased Hb (7.8 g /dl). CT revealed huge hematoma in the second portion of duodenum with dilated bile duct. An emergency endoscopy showed huge hematoma but the sutured site was intact, and there was no active bleeding at this point. Hematoma was removed endoscopically, and i
t took approximately 5 hours. Thereafter, hepatobiliary pancreatic enzymes improved. Oral intake was started on POD13, but the next day, she had melena again with a decrease in Hb. An endoscopy was performed again. There was an exposed vessel within a duodenal ulcer near the third portion of duodenum, and hemostasis was achieved by ablation. The endoscope could not pass through the sutured bulbus due to stenosis, but the bypass route was very supportive for this procedure. After that, the patient&#039;s course was generally stable, and she was discharged on POD25.
[Discussion]
This is a rare case of papillary obstruction due to hematoma in duodenum. The performed operation was on emergency to save the life, but we added the bypass of gastrojejunostomy just in case. Eventually, this bypass route was very effective for endoscopic hematoma removal with hemostasis and oral intake after surgery.
[Conclusion]
We report a case of duodenal perforation and postoperative papillary obstruction due to hematoma that was saved by emergency surgery and endoscopy.

Event date： 2023.11

Venue：Kobe   Country：Japan  

Serous cystic neoplasms (SCNs) are pancreatic tumors characterized by clusters of small cysts filled with serous fluid. The majority of them are benign, however, surgical resection of them may be considered in cases where the tumors are increasing in size, causes any symptoms, or where malignancy is suspected. SCNs are highly vascular tumors due to numerous blood vessels around the cystic legions. In addition, larger tumors cause obstructive pancreatitis and vascular adhesion with inflammation, therefore surgery is often difficult. Here we report a case of pancreatic resection for a giant hypervascular pancreatic serous cystic tumor in which the tumor feeding vessels were embolized preoperatively.
The patient is a 60-year-old male. An 8-cm tumor was found in the body of the pancreatic head on CT and was diagnosed as a SCN by endoscopic ultrasound guided tissue acquisition. The tumor was a microcystic type, exhibiting extremely abundant vascular development within the tumor, and vein expansion around the tumor was observed. We decided to perform surgical resection due to its enlargement by 16mm over the course of 7 years. To control intraoperative bleeding, the distal end of the gastroduodenal artery (anterior and posterior supra pancreaticoduodenal artery) and the dorsal pancreatic artery branching from the superior mesenteric artery, which are tumor feeding vessels, were embolized using Interventional Radiology on the day before surgery. The pancreatoduodenectomy was completed with an operating time of 5 hours and 15 minutes and a blood loss of 763 ml, with no uncontrolled bleeding. The postoperative course was uneventful and the patient was discharged without complications.
Although reports of preoperative vascular embolization for pancreatic tumors are extremely rare, this strategy was considered to be extremely useful for safe surgery.

Event date： 2023.10

Venue：Yokohama   Country：Japan  

Background: Gastrointestinal neuroendocrine carcinoma (NEC) is a rare tumor originated from neuroendocrine cells. Multidisciplinary treatment centered on chemotherapy is performed, but NEC is highly malignant and the prognosis is extremely poor. Treatment strategies of gastric MALT lymphoma differ depending on the positivity of Helicobacter pylori. If eradication is unsuccessful, the treatment shifts to chemotherapy and radiation, but if the patient is asymptomatic, keeping observation is also considered. Herein, we report a case of gastric NEC combined with MALT lymphoma who underwent preoperative chemotherapy and was successfully resected.
Clinical course: A 69-year-old woman who had a chief complaint of epigastralgia visited our hospital. Endoscopy showed an ulcerative lesion in the lesser curvature of the lower gastric body, and a biopsy revealed NEC. On the other hand, she had an edematous mucosa from the upper to the lower gastric body, and biopsy revealed MALT lymphoma. As for tumor markers, NSE and ProGRP levels were elevated, and CT showed enlarged perigastric lymph nodes, suggesting lymph node metastasis. Eradication therapy was unsuccessful. The primary tumor and metastatic lymph nodes were shrunk by 3 courses of chemotherapy of etoposide and cisplatin (EP therapy). Then, we performed laparoscopic distal gastrectomy with D2 lymph nodes dissection as radical resection for the gastric NEC. On pathological examination, there was no tumor cells in either the primary tumor or lymph nodes, and was diagnosed with complete remission (CR). Moreover, no MALT component was observed in the specimen. Two courses of EP ther
apy were added as adjuvant chemotherapy, and no recurrence has been observed.
Conclusion: In this case, chemotherapy for NEC was highly effective, and a radical resection and CR were obtained. It is necessary to accumulate more cases to provide the evidence of adequate treatment for NEC.

Event date： 2023.10

Venue：横浜市   Country：Japan  

Due to its high effectiveness, heavy ion radiotherapy in particular has been applied to advanced medical
treatment since 2003, covered by insurance for some cancers since 2016, and has expanded further since 2022,
becoming one of the pillars of cancer treatment. Due to its high dose distribution during heavy ion radiotherapy,
a distance is required between adjacent organs, and a spacer may be required. We investigated the spacer surgery
performed in our department. From 2013 to 2023, we investigated 40 patients who underwent spacer placement
for the indication of heavy ion radiotherapy. Fourteen males and 26 females. Their average age is 58.3 (22-85).
Two cases were resectable based on intraoperative findings. Thirteen patients of recurrence of rectal and uterine
cancer, 17 patients of sarcoma, and eight patients of sarcoma recurrence underwent spacer placement. The
procedure was Gore-Tex sheet placement in 23 patients, absorbable pacer placement in 9 patients, omental filling
in 3 patients, and pelvic floor reconstruction in 3 patients, and spacer placement was possible in all patients. As a
short-term complication, one patient had an intractable pelvic abscess with a Gore-Tex sheet, and one patient had
a duodenal perforation due to the Gore-Tex sheet. No such postoperative complications were observed with
resorbable spacers. We experienced a case in which an absorbable spacer was placed at the same time as
ileocecal resection because of obvious and inseparable adhesion between tumor and ileocecum. Although no
infection was observed in this case, there are reports of cases in which an absorbable spacer has become infected,
so placement of a spacer should be avoided in contaminated surgery. Number of the patients of spacer placement
will increase further with insurance coverage for x-ray radiation therapy using absorbable spacers. It is very
important to determine whether the spacer can be placed safely and the tumor can be resected.

Event date： 2023.10

Venue：横浜市   Country：Japan  

Introduction:
Ureteral injury in colorectal surgery is sometimes diagnosed postoperatively and could be fatal,
which may cause a decreased quality of life. Although anatomical recognition during surgery is
important, it is difficult to identify the accurate anatomy on intraabdominal severe adhesions.
Fluorescent ureteral catheter (NIRC) could make the ureteral recognition easy during laparoscopic
surgery with its fluorescent dye like indocyanine green (ICG). We herein report a case with
laparoscopic surgery for metachronous residual sigmoid colon cancer in which NIRC was useful to
identify the ureter.
Case report:
A 74-year-old man underwent laparoscopic sigmoidectomy with D2 lymph node dissection by
medial approach for sigmoid colon cancer (pStage I) 3 years ago. The colonoscopy for postoperative
surveillance revealed advanced cancer in the residual sigmoid colon. We planned to perform a
laparoscopic high-anterior resection including previous anastomotic site. There was concern about
left ureteral injury because of the previous left-side colon mobilization, thus we inserted a NIRC to
left ureter just before the operation. Intraoperative findings showed adhesion between the left-side
mesocolon and retroperitoneum as expected, while visualization of the ureter by NIRC lead us to
recognize the ureter and dissection layer as appropriate, and NIRC made possible to mobilized the
colorectum from splenic flexure to peritoneal reflection by medial approach again. The operation
time was 211 minutes and blood loss was minimal. There was no adverse event associated with
ureteral catheter insertion. The patient was discharged on the 15th hospital day without any
complication.
Summary:
Recently, ICG-based navigation techniques have been shown to be effective in endoscopic surgery.
Although there are some issues to solve such as postoperative hematuria, prolonged operation time,
and higher cost, NIRC was considered useful for intraoperative identification of the ureter during
endoscopic surgery.

Event date： 2023.9

Venue：福岡市   Country：Japan  

食道平滑筋腫は良性腫瘍のため経過観察されることが多いが、サイズが大きくなるとつかえ感などの症状が出現し、手術適応となる。標準術式は腫瘍核出術であり、低侵襲である鏡視下手術が考慮される。さらなる低侵襲手術として経口内視鏡的腫瘍核出術(POET)が開発されたが、腫瘍径による制限がある。我々は食道平滑筋腫に対するPOET併用による胸腔鏡下手術（TECS）を3例経験した。年齢は32歳から48歳であり、男性1名、女性2名であった。１例目は胸部下部食道に亜全周性の60mm大。２例目は胸部中部食道に70mm大。３例目は門歯27cmから34cmまでの胸部食道に最大径40mmの平滑筋腫を３個認めた。いずれの症例も、全身麻酔下仰臥位にてまず経口内視鏡を用いて腫瘍に至る粘膜下トンネルを作成し、腫瘍周囲の剥離を完了させた。続いて腹臥位にて腫瘍周囲の食道を剥離受動し、外膜筋層を切離して腫瘍を核出した。筋層の欠損部を縫合閉鎖し、内視鏡にてentry holeをクリップ縫合し手術を終了した。手術時間は273分から392分で、出・オ樊フ未1gから25gであった。いずれの症例も術中・術後合併症なく経過良好であった。また狭窄や機能障害もなく外来経過観察中である。本手術は、内視鏡的に腫瘍周囲剥離を先行することで、サイズが大きく不整形な食道粘膜下腫瘍や多発例に対しても、安全かつ確実な手技であると考え、発表する。

Event date： 2023.8

Venue：東京都   Country：Japan  

【目的】膵癌はKRASやTP53など多くの変異が蓄積していることが知られているが、リキッドバイオプシーを用いた早期発見は未だ困難である。発癌から癌進展に至るまでの遺伝子変異をリキッドバイオプシーを含め解析し、各段階での重要な遺伝子変異を見つけようと考えた。また、これらの重要な変異が腫瘍微小環境に与える影響についても検討する。
【方法】KPCマウスの末梢血を毎週採取しゲノムDNAを抽出した。Sacrifice時には末梢血と血性腹水からCTCsを分離しゲノムDNAを抽出した。膵原発および肝転移のFFPEからも同様にゲノムDNAを抽出した。各ゲノムDNAを用い、NovaSeqでエクソームシークエンスを行った。
【結果】解析に用いたKPCマウスを、原発なし、原発あり/転移なし、原発あり/転移ありの3群に分けた。経時的な末梢血中のCTCsと膵原発のFFPEとで遺伝子変異を比較したところ、2つの変異が検出され、そのうち1つはMHCクラスIに関連していた。一方、原発なし群ではMHC I変異は認められなかった。原発あり群の膵原発組織では原発なし群と比較してCD8+T cellsクラスターが領域リンパ節に見られた。発癌初期から現れたMHC Iを含む2つの変異は末梢血と血性腹水において優位であったが、肝転移では検出されなかった。
【結論】MHC I変異は発癌初期から血中を循環しており、早期発見の指標となる可能性がある。免疫応答によりMHC I変異を持つ癌細胞が肝転移において除去される可能性が示唆された。

Event date： 2023.7

Venue：鹿児島市   Country：Japan  

憩室炎は良性疾患であり低侵襲手術が望ましいが腹腔内の炎症や癒着などを伴う複雑性憩室炎に対する鏡視下手術の安全性及び妥当性はいまだ明らかでない。膀胱との瘻孔を伴う憩室炎に対する腹腔鏡手術の短期成績を検討し、安全性及び妥当性を検証する。2012年~2022年に手術施行した膀胱瘻を伴う憩室炎15症例を対象とした。男性7例女性8例、年齢60.5(35-74)歳、罹患範囲はS状結腸8例、下行~S状結腸6例、全結腸1例であった。術前検査の瘻孔診断能はそれぞれCT 8/15 (53.3%)、消化管造影3/11 (27.3%)、MRI 7/7 (100%)とMRIが瘻孔診断能に優れていた。全身状態から人工肛門造設のみ行った1例を除いた全例において腹腔鏡で手術を開始したが2症例において腹腔内の著明な炎症癒着により開腹に移行した。人工肛門は回腸3例、Hartmann２例造設したが全例で閉鎖可能であった。鏡視下手術における術中の工夫としては腹腔鏡の下からのぞき込むように見ることができる視野を利用して瘻孔部の周囲、特に背側や奥の剥雷オ档C鮴莵圓靴峠侏茲襪世閏・阿鮃圓辰晋紊磨餽ι瑤鮴擲・垢襪海箸煤餽Δ魴狙・靴紳ヾ錣梁蚕・鮑脳・造砲任④襪隼廚錣譴襦

Event date： 2023.7

Venue：福岡市   Country：Japan  

【背景・目的】膵癌は非常に予後不良な癌の一つであり、過剰な間質増生とオートファジー依存的な腫瘍増殖が特徴である。膵癌ではオートファジーの亢進が化学療法抵抗性や予後の悪化と相関していると報告されているが、膵癌のオートファジーを制御する有効な治療法は見つかっていない。我々は膵癌微小環境を構成する膵星細胞のオートファジーが膵癌の浸潤・増殖に与える影響について報告してきた。本研究では膵星細胞の研究で得られた知見をもとに、膵癌細胞のオートファジーを制御する遺伝子を明らかにし、膵癌の浸潤、増殖に与える影響について検討する。
【方法】膵臓癌、慢性膵炎患者より得られる手術切除標本からそれぞれ膵星細胞を樹立した。これらをマイクロアレイに提出し両者の遺伝子発現を比較した。膵臓癌患者の膵星細胞に高発現する標的遺伝子の中から膵癌細胞にも発現する遺伝子を同定し、膵癌細胞におけるオートファジーや浸潤・増殖に与える影響について検討した。
【結果】遺伝子発現解析の結果、膵癌患者から樹立した膵星細胞にはendoplasmic reticulum aminopeptidase 2(ERAP2)が高発現しており、オートファジーを制御する遺伝子と考えられた。ERAP2はSUIT-2やBxPC-3などの膵癌細胞株にも発現していた。ERAP2をノックダウンするとこれらの膵癌細胞株のオートファジーが阻害され、増殖能が抑制された。ヒト膵癌手術検体FFPEの免疫染色では、腫瘍内ERAP2の高発現は予後不良と関連していた。
【考察】膵癌細胞内に発現するERAP2はオートファジーを制御しており、膵癌の浸潤・増殖に関与していることが示唆された。これらの結果よりERAP２は膵癌における新たな治療標的となり得ると考えられた。

Event date： 2023.7

Venue：福岡市   Country：Japan  

【背景と目的】
難治性腫瘍の代表である膵癌において抗腫瘍免疫療法の有効性は示されていない。近年、膵癌の免疫抑制性微小環境を形成する重要なファクターとしてDC (Dendritic cell) の機能低下が報告されている。我々は、細胞内小器官を分解する代謝機構であるAutophagyに着目し、Autophagy活性が持続的に高い膵癌細胞では、抗原となり得るタンパクが分解されることがDCの機能低下につながっているのではないかと考えた。DCの活性化に着目したAutophagy抑制による新たな腫瘍免疫増強のメカニズムを解明し、Autophagyを標的とした新規複合免疫療法の可能性を追求することが本研究の目的である。
【方法】
1. 膵癌自然発癌マウスモデル(KPCマウス)より樹立した膵癌細胞株を用いてAutophagy関連遺伝子であるATG7をノックダウンした細胞株、KPC shATG7(Autophagy抑制細胞株)を作成した。KPC shNC(コントロール)とKPC shATG7をそれぞれC57BL/6マウスに皮下移植し、腫瘍形成の比較と腫瘍内免疫細胞の解析を行った。同様の実験をBALB/c-nuマウス(T細胞欠損免疫不全マウス)でも行った。
2. KPCshNC、KPCshATG7の膵同所移植腫瘍における免疫微小環境をSingle-cell RNA-seqにより解析した。
3. KPC由来膵癌細胞とマウス骨髄由来DCを共培養し、癌細胞のAutophagyを抑制した際のDCの活性化と抗原提示能の変化をFACSで検証した。
【結果】
1. KPC shATG7群で有意に腫瘍縮小を認め、腫瘍内CD8+Tcellの増加とDCの活性化マーカーの発現上昇を認めた。さらに、BALB/c-nuマウスでも、活性化したDCが増加しており、Tcellによる細胞障害性とは独立して、癌細胞のAutophagyとDC活性化の直接的な関係が示唆された。
2. scRNAseq解析の結果、shATG7群ではDC2, migratoryDCのクラスターにおいて抗原提示能、DC成熟化、遊走能に関連する遺伝子群の発現が上昇していた。
3. Autophagy を抑制した癌細胞と共培養したDCはCD80, CD86, MHC class ?/?の発現が上昇した。さらにAutophagy抑制癌細胞と共培養したDCは、CD8+T細胞との共培養で、T細胞の増殖を促進させた。
【考察】
膵癌細胞のAutophagyを抑制することでDCの機能が増強され、T細胞primingが促進されることがin vitro, in vivoの実験から示唆された。

Event date： 2023.7

Venue：函館市   Country：Japan  

背景：膵・胆管合流異常を伴う先天性胆道拡張症に対する手術は，発癌母地である拡張胆管を切除して膵液逆流を防止し，症状と発癌を予防することが目的である．遺残した膵内胆管は膵石や癌が発生することがあるものの，膵管損傷などの重大な合併症を生じることがあるため，術前に合流形態を診断し剥離範囲を決めておく必要がある．また，局所解剖がわかりにくい場合にはLillyら (Surg Gynecol Obstet. 1978; 146: 254-6)が推奨した拡張胆管壁内での剥離が安全であり，ロボット胆管切除術でも有用か検証した．
患者と方法：2006年4月から2022年11月までに当科で先天性胆道拡張症に対して鏡視下胆管切除を行った39例を対象とした．膵内胆管では胆管壁内で剥離を行うことを基本とし，ERCP/MRCP画像を元に，膵・胆管合流異常を分類し（Type A 狭窄型，Type B 非狭窄型，Type C 共通管拡張型，Type D 複雑型，膵・胆管合流異常研究会診断基準検討委員会による分類)，膵内胆管の完全切除が可能と考えられるType A 狭窄型における遺残胆管の有無を術中所見・術後フォローアップCT, MRIで確認し，術式（腹腔鏡 vs ロボット）ごとの結果を検証した．
結果：鏡視下胆管切除術の術式は腹腔鏡28例，切除腹腔鏡/再建ロボットが4例，完全ロボットが7例であった．Type A 狭窄型は腹腔鏡で胆管切除を行った32例中，16例 (50%)，完全ロボット胆管切除では5例中7例 (70%)であった．Type A狭窄型の内，膵内に遺残胆管を認めなかったのは腹腔鏡切除で6例 (6/16, 38 %)，ロボット切除で1例 (1/5, 20 %)であった(P=0.62)．主膵管損傷はなく，術後膵液瘻は腹腔鏡切除の1例 (1/16, 6.3%)で認められた．
結論：Type A 狭窄型の膵・胆管合流異常における膵内胆管の遺残は腹腔鏡，ロボット切除で有意差を認めなかったものの，ロボットの多関節機能は胆管壁内での剥離を容易にし，安全な膵内胆管切除に有用であると考えている．腹腔鏡・ロボットでのそれぞれの手技をビデオで供覧する．

Event date： 2023.7

Venue：函館市   Country：Japan  

【背景】胃癌治療において免疫チェックポイント阻害薬の導入はこれまでの治療に画期的な変化をもたらした．抗PD-1抗体による治療において，免疫染色で評価されるCPSは薬剤投与の指標として期待されている．しかし，腫瘍免疫におけるCPSの意義は未だ明らかではない．
【目的】胃癌症例において，CPS 高低値に分類しシングルセル解析を用いてCD8陽性T細胞の機能を詳細に解析し，微小腫瘍免疫環境におけるCPSの意義を明らかにする．
【方法】当外科で胃切除手術を施行した11症例の胃癌患者から腫瘍部11サンプルを採取し，各症例の切除組織を対象に抗PD-L1抗体及び抗CD8抗体を用いて免疫染色を行い，CPS分類及びCD8陽性細胞数の計測を行った．また，single cell RNA-sequence解析を行い，CD8陽性T細胞をサブセットとして抽出し，CPSによって低中高値の３分類に分け， CD8陽性T細胞の遺伝子発現から推測される機能との関連性を評価した．
【結果】胃癌全11症例の免疫染色においてCPSとCD8陽性T細胞数は正の相関を認めた．各々の症例に対してシングルセル解析データの統合解析を行い，CD8陽性T細胞を抽出した．対象細胞数は，10610細胞で，5つのサブクラスターに分類できた。CPS高値群では、CD8陽性T細胞の総数は他の群と比較して多く，NaiveT細胞の割合は低く，疲弊化マーカーを発現したCD8陽性T細胞の割合が高かった．また，CPS高値の群では，CD8陽性T細胞において，低酸素関連遺伝子が有意に発現していた．
【結語】腫瘍免疫におけるCD8陽性T細胞の機能は、腫瘍や免疫細胞のPD-L1発現相関していることがわかった。また、CPS高値の腫瘍ではCD8陽性T細胞は疲弊化しており，低酸素環境に置かれていることが示唆された.

Event date： 2023.7

Venue：函館市   Country：Japan  

Event date： 2023.7

Venue：函館市   Country：Japan  

【背景】胃癌・食道胃接合部癌において、非充実型低分化型腺癌 (por2)は予後不良であり、一般的に豊富な繊維化組織を伴うことが多い。近年、膵癌や乳癌でcancer-associated fibroblast (CAF)のheterogeneityがシングルセルRNAシーケシング (scRNA-seq)によって解明されているが、por2胃癌・食道胃接合部癌微小環境におけるCAFとCD8陽性T細胞の相互作用をscRNA-seqを用いて解明した報告はない。
【対象】当科で根治切除術を施行した全21例から採取した胃癌19例 (por2: 9例)と食道胃接合部癌2例 (por2: 1例)を対象とした。
【結論】全21症例の統合解析を行った結果、quality check後の癌関連線維芽細胞 (CAF)の細胞数は3338であり、遺伝子発現が異なる5つの集団に分類された。Por2以外の胃癌・食道胃接合部癌と比較してpor2胃癌・食道胃接合部癌で多いCAFの細胞集団はACTA2・TAGLN・MMP11の発現が高値であること、Gene Ontology解析で「actin filament-based process」「Smooth Muscle Contraction」のアップレギュレートを認めたことから、Myofibroblastic CAFと判断した。また、CD8陽性T細胞はPor2以外の胃癌・食道胃接合部癌と比較してpor2胃癌・食道胃接合部癌で有意に少なく(P = 0.03)、細胞障害機能関連遺伝子の発現も有意に低値であった (P < 0.001)。
【結論】 por2胃癌・食道胃接合部癌のCAFにはheterogeneityがあり、Myofibroblastic CAFが腫瘍内へのCD8陽性T細胞浸潤を抑制している可能性が示唆された。

Event date： 2023.7

Venue：函館市   Country：Japan  

【背景】食道扁平上皮癌 (ESCC)は予後不良な消化器癌の一種であるが、免疫チェックポイント阻害薬 (ICB)の有効性が報告され腫瘍免疫微小環境の理解が急務となっている。近年、TIME内の3次リンパ組織様構造 (TLS)と呼ばれる構造物が注目されている。TLSは腫瘍局所などに後天性に形成される二次リンパ組織と類似した構造物でありTリンパ球、Bリンパ球、樹状細胞、マクロファージなどの免疫細胞や、細網線維芽細胞などの間質細胞から構成される。またTLSの内部には胚中心が形成され活発な免疫応答が行われていると考えられている。腫瘍内のTLSの存在はICBの奏効率を予測し、乳癌・肝細胞癌など多くの固形癌の良好な予後と相関していることが報告されているがESCCにおける報告は少なくその臨床的意義は不明な点が多い。
【目的】ESCCにおけるTLSの臨床病理学的意義を明らかにする。
【対象】2008年4月～2020年11月の期間に当科で切除術を行った食道扁平上皮癌180例を対象にTLSと臨床病理学的因子や予後との関連を評価した。TLSはH&E染色と抗CD20抗体を用いた免疫組織化学染色で評価した。統計解析はLog-rank test、生存時間分析はKaplan-Meier法を用いた。
【結果】年齢中央値は63歳(41-83歳)、男性は153例 (85%)、女性は27例 (15%)であった。TLS+群は144例 (80%)、TLS-群は36例 (20%)であった。臨床病理学的因子に関して、TLSの存在とT1/2 (p = 0.0196)、所属リンパ節への転移がないこと(p = 0.0294)、pStageI/II (p = 0.0029)は有意に相関していた。一方で年齢や性別には有意な相関は認められなかった (p = 0.795、p = 0.877)。またTLS+群はTLS-群と比較して、recurrence free survival/overall survivalの延長に有意に相関していた(p < 0.0001, p = 0.0016)。
【結語】ESCCにおけるTLSの存在は良好な予後予測因子と考えられた。TLSは抗腫瘍免疫を増強している可能性が示唆され今後ICBをはじめとした個別化治療を発展させるための重要な知見となりうると考えられる。

Event date： 2023.7

Venue：函館市   Country：Japan  

Event date： 2023.7

Venue：函館市   Country：Japan  

【緒言】
切除不能胃癌に対してTrastuzumabを併用した化学療法が適応となり, 化学療法が奏功しConversion surgeryを行った報告が散見されるようになった. しかし, 周術期の抗HER2療法においては複数の臨床試験が進行中であり, 未だcontroversialである. 今回我々は遠隔転移を伴うHER2陽性胃癌に対して抗HER2療法が奏功しConversion surgeryを行った症例及び術後化学療法で抗HER2療法を行った症例を経験したため若干の文献的考察を踏まえて考察する.
【症例1】
患者は55歳男性. 多発の腹膜播種を伴った切除不能胃癌に対してHER＋SP[Herceptin＋S-1＋Cisplatin]療法を7コース施行したのち, 画像上で播種病変が消失したため審査腹腔鏡を施行したところ腹膜に瘢痕組織を複数認めた. 複数箇所生検を行うも全て瘢痕組織であり, 化学療法が奏功したと判断して後日腹腔鏡下胃全摘術[D2, Roux-en Y再建]を施行した[Type4, ypT3, N3a, M, ypStage?B]. 術後補助化学療法としてS-1＋DTX[S-1＋Docetaxel]療法を約1年施行したが, 術後2年で多発リンパ節転移を認めた. その後Herceptinを含めたレジメンを各種行うもPDとなり, 最終的に骨髄癌腫症を発症し術後約3年で逝去した.
【症例2】
患者は75歳男性. 進行胃癌に対して腹腔鏡下胃全摘術[D2/下縦隔郭清, Roux-en Y再建]を施行した[Type3, pT4a, N3a, M0, pStage?B]. 免疫染色ではHER2（3＋）であり, 術後化学療法としてHER＋XP[Herceptin＋Capecitabine＋Cisplatin]療法（有害事象のため2コース目以降はcisplatin中止）を8コースとCapecitabineを16コース施行した. 術後2年で単発性の脳転移を認め,開頭腫瘍摘出術を施行した. 術後病理結果ではHER2の発現はequivocal[2＋]と原発巣と比較して低かったが, 原発巣の病理像に類似しており胃癌の転移性腫瘍と判断した. 今後は頭蓋内奏効率など考慮しニボルマブ単独投与で再発治療を行う方針となり, 現在無再発経過中である.
【考察】
HER2陽性の切除不能胃癌の全生存期間は約16ヶ月とされており, 症例1ではConversion surgeryを行えたことが約4年という予後に寄与した可能性がある. また, 症例2では術後の抗HER2療法を含めた化学療法により頭蓋内以外の病勢はコントロール良好であったが, 予後不良とされる胃癌脳転移で再発し, 頭蓋内における病勢コントロールは不良であったと考えられる. 抗HER2療法を含めた集学的治療に関しては今後のさらなる症例の蓄積が必要である.

Event date： 2023.7

Venue：函館市   Country：Japan  

【背景と目的】
切除可能膵癌(R膵癌)に対して術前治療（NAT）を行うことが提案されている一方で、R膵癌患者全てにおいてNATが有効であるかなど不明な点も多く、今回R膵癌に対するNATの意義や適応に関して検討することを目的とした。
【方法】
2012年から2021年に術前にR膵癌と診断、手術を施行し最終病理で浸潤性膵管癌と診断された、切除先行群（UF群）：195例、NAT群:83例を対象として臨床病理学的因子や長期予後に関して後方視的に検討を行った。さらに術前に得られる因子から予後予測因子を検討し、NATがより有効と思われる患者の検討を行った。
【結果】
R膵癌全体において、2年生存率（UF群 vs NAT群: 65.4% vs 74.6%, p=0.137）や2年無再発生存率 (UF群 vs NAT群: 48.7vs 61.5%, p=0.119）には両群で有意差は認めなかった。術後病理学的因子に関してR0切除率はNAT群で高い傾向（UF群 vs NAT群: 89 vs 95%, p=0.094）にあった。術後合併症率（CD≧3）（UF群 vs NAT群: 11 vs 13%, p=0.557）は有意差を認めず、術後補助化学療法の施行率（UF群 vs NAT群: 81 vs 88%, p=0.173）に関しても両群において有意差は認めなかった。
R膵癌切除症例における術前に同定可能な因子における多変量解析では、腫瘍径≧2cm、CA19-9>100、アルブミン/グロブリン比<1.38が独立した予後不良因子であり、これらの３項目からlowリスク(0/1点)(n=156)、highリスク(２/3点)(n=122)に分類を行い、予後を比較検討したところ、highリスク症例においては2年生存率（UF群 vs NAT群: 45.1% vs 70.6%, p=0.048）はNAT群がUF群と比べて有意に良好な結果であった。lowリスク症例においては2年生存率（UF群 vs NAT群: 79.3% vs 77.0%, p=0.965）では両群で有意差は認めなかった。
【結論】
R膵癌に対するNATの意義に関して、highリスク症例においてはNATが有効である可能性が示唆されるが、lowリスク症例におけるNATの意義を含めて今後のさらなる検討が必要である。

Event date： 2023.7

Venue：函館市   Country：Japan  

＜目的＞
2020年に保険収載されたロボット支援下膵頭十二指腸切除術（RPD）は多くの施設が未だ、導入期であると思われる。2020年に開催されたコンセンサスミーティング（PAM meeting）では右側からのアプローチが基本的手技として知っておくべきであるというコンセンサスが得られた一方、様々な症例に対応するためにもアプローチ方法に関してはvariationを持っておくべきであるとの意見もあった。右側からのアプローチでは空腸も右側から引き抜く事は可能であるが、Treitz靭帯がその妨げとなることが多く、当科では左側からのアプローチも行なっている。今回、当科におけるRPDの視野展開、手術手技をビデオで供覧し、また、RPDの安全性と有用性を検証する。
＜方法＞
当科では2018年に臨床試験下にRPDを開始、当科では以前より開腹手術同様、左側からのアプローチも併用している。手術開始後体位は頭高位10度、必要に応じて右側に5度傾ける。空腸を足側、右側に展開しTreitz靭帯、切離、このとき十二指腸水平脚に付着するTreitz靭帯は背側から切離を行うことで完全に切離を行うことが可能となる。必要に応じて左側から膵頭部背側の剥離も行う。本操作によりRPDでも右側から空腸を抵抗無く容易に引き抜くことが可能となる。その後右側からのアプローチを行い、膵頭部をSMA、SMVから切離し標本を摘出する。
＜結果＞
2022年11月までに行った54例のRPDを解析対象とした。手術時間は1-10、11-20、21-30、31-40例目の手術時間および出血量中央値はそれぞれ、776分（689-846分）、746分（665-881分）、784分（542-998分）、679分（581-830分）であり、出血量中央値は354g（80-1550g）、294g（124-1950g）、290g（17-1330g）、175g（50-612g）であった。一方-41例目以降の手術時間中央値、出血量中央値は581分（437-855分）であり、201g（0-1050g）であった。また、膵液瘻発生率に関しては10例（21.3%；全例soft pancreas）であり、周術期死亡は認めていない。
＜結語＞
当科では症例の拡大に伴い手技の変遷を行い、現在までにRPDを安全に導入できていると考える。

Event date： 2023.7

Venue：函館市   Country：Japan  

＜はじめに＞
膵癌に対する治療は膵癌診療ガイドラインが2019年に改訂されて以降、術前・術後補助化学療法が標準治療となった。膵癌患者の予後改善には、集学的治療が不可欠であり、できるだけ少ない侵襲で手術を行い、速やかに術後補助化学療法につなげることが重要である。そのため、膵癌に対する低侵襲手術（MIS）が期待されており、腹腔鏡下膵体尾部切除術（LDP）は保険収載されて以降、広く普及しつつある。それに伴い、肥満や進行癌など、困難症例に出会うこともある。そのような場合、一つではなく様々なアプローチに精通することで対応が可能となる。当科では、LDPを施行する際、膵後方の剥離を頭側からアプローチすることが多い。当科におけるLDPの手術手技を供覧しながら概説する。
＜手術手技＞
総肝動脈を確保後、左胃動脈左側から胃膵ヒダ背側を剥離する。Gerota筋膜前面の疎な層は容易に剥離可能である。胃膵ヒダを脾上極に向けて切離する。助手の左右の鉗子で切離した胃膵ヒダを足側に牽引すると安定した視野でGerota筋膜および同膜に覆われた左副腎を透見できる。Gerota筋膜を切開し、膵尾側へ向けて膜を意識しながら剥離を続ける。腹腔鏡の鉗子の軸がGerota筋膜切開の方向に一致するため、展開を変えずに膵尾部まで進むことができる。剥離が進むと脾動脈根部を切離し、助手がGerota筋膜を把持して足側へ牽引することで、さらに膵の授動を行うことが可能となり、視野を変えることなく左副腎静脈、左腎静脈前面を露出、さらには膵下縁付近まで剥離可能である。
＜結果＞
2016年から2022年の間に浸潤性膵管癌に対して施行したLDPは36例であった。手術時間中央値、出血量中央値はそれぞれ303分（180 - 535分）、200ml（0 - 1900ml）であった。R0切除率は97%であった。膵液瘻（Grade B/C）、胃内容排出遅延（Grade B/C）、Clavien-Dindo III以上の合併症頻度は、それぞれ19%、6%、16%であり、術後在院日数中央値は16日（10 ? 120日）であった。
＜結語＞
頭側からのアプローチは膵癌に対するLDPにおけるアプローチの一つとして有用であると思われる。

Event date： 2023.7

Venue：函館市   Country：Japan  

Event date： 2023.7

Venue：函館市   Country：Japan  

食生活の変化などによって本邦では大腸憩室疾患が増加している．憩室の原因として腸管や膀胱は比較的多く経験する
が，憩室炎が原因の子宮や膣などの婦人科臓器への瘻孔は比較的少なく，その臨床病理学的特徴及び腹腔鏡手術に対す
る安全性については明らかでない．今回我々は2007年から2021年に手術を施行した瘻孔を伴う憩室炎症例22例のうち婦
人科臓器への瘻孔を認める5例について検討した．同時期の瘻孔を伴う憩室炎では13例の膀胱瘻，4例の小腸瘻を認め
た．5例全例でS状結腸の憩室が瘻孔の原因となっており，瘻孔の形成先は4例が膣，1例が子宮であった．膣への瘻孔を
認めた4例は全例で子宮全摘後の膣断端に瘻孔部を認め，子宮全摘後のS状結腸憩室は同部への瘻孔のリスクが高いこと
が示唆された．画像による診断では結腸膣瘻の正診率はCT2/5 (40.0%), 注腸 1/4 (25.0%)であった．以前の我々の検討で
は結腸膀胱瘻の瘻孔同定にはMRIが最も鋭敏な検査であることが示されているが今回の結腸膣瘻症例に対してはMRIは
1例も施行されていなかった．術式はS状結腸切除1例，前方切除2例，Hartmann手術1例，ストーマ造設1例でそのうち2
例に対して腹腔鏡手術を施行した．手術時間は365分，出血量は55mlであった．CD2以上の術後合併症は脳梗塞1例，腹
腔内膿瘍1例であった．全例に術中内診を施行して瘻孔部を確認した．消化管再建はDST2例，手縫い1例で施行した．
膣の欠損部は全例単純閉鎖を行った．切除標本は組織学的には全例活動性炎症を認め，1例に膿瘍を認めた．2例
(50.0%)で組織学的に結腸膣瘻が証明された．今回の結果で憩室炎による結腸膣瘻などの婦人科臓器への瘻孔に対する
腹腔鏡手術は腹腔鏡手術に習熟したチームでは安全に施行できることが示唆された．

Event date： 2023.7

Venue：函館市   Country：Japan  

Event date： 2023.6

Venue：大阪市   Country：Japan  

Event date： 2023.6

Venue：Seattle  

Purpose
Familial colorectal cancer (CRC) is a risk factor for CRC in certain families. However, its prognostic impact in CRC patients remains controversial.The study aimed to clarify the prognostic impact of CRC family history.
Methods
We retrospectively reviewed the database from 1978 until 2018 and enrolled 3,655 consecutive CRC patients in this study. We investigated the clinicopathological factors of CRC patients with and without family history. After propensity score matching, we performed a survival analysis of CRC patients with and without a family history.
Results
CRC patients with a CRC family history had a young onset (63.2 and 65.9; P<0.001), were more likely to be female (54.3% and 49.7%; P=0.042), had less symptomatic disease (76.9% and 80.8%; P=0.008), were more likely to have right-sided colon (27.5% and 26.1%), and had less distant metastasis (11.3% and 14.9%; P=0.023) and more multiple CRCs (10.2% and 7.8%) compared with those without a CRC family history. Prior to propensity score matching, colorectal cancer-specific survival analysis showed that CRC family history was a good prognostic factor (P=0.022). After propensity score matching, survival curves were overlapping between the two groups.
Conclusion
CRC patients with a CRC family history had specific clinicopathological features including younger onset, female sex, proximal colon location, fewer symptoms, smaller number of distant metastases, likely to be a multiple disease and earlier cancer stage. Family history of CRC in CRC patients was not prognostic factor itself.

Event date： 2023.5

Venue：名古屋市   Country：Japan  

Event date： 2023.5

Venue：名古屋市   Country：Japan  

本邦ではロボット支援下膵頭十二指腸切除術（RPD）、尾側膵切除術（RDP）が2020年に保険収載され、全国的にも症例数徐々に増加傾向である。当科では2018年に倫理委員会承認のもとRPDを開始、2021年1月までに102例のロボット支援下膵切除術を行ってきた（RPD;60例、RDP;42例）。適応に関しても当初は良性～低悪性腫瘍に対して行なってきたが、徐々に適応を拡大し、現在ではRDP,RPD共に浸潤性膵癌に対してもおこなっている。今回膵癌に対するRPDの手技をビデオで供覧し、手術成績を報告する。膵癌に対するRPDでは随伴性膵炎および上腸間膜動脈や門脈周囲への癌の進展により手技的に難易度が高いと考えられている。我々は安全に行うために徐々に手技を変遷させてきた。SMA左側はSMA右側に比べて炎症の程度は低いと考えている、そのため、膵癌に対してはSMA左側からのアプローチを先行して行なっている。左側からのアプローチでは始めに横行結腸間膜を頭側に翻転、空腸を扇状に展開、空腸起始部でTreitz靭帯を完全に遂オ椌フイ靴討・・△修慮紂・・牡嵋譴鮴擲1st JA、2nd JAさらには1st JVを確認する。その後これらの血管をSMA左側で切離したのちに小腸間膜を切開する。その後、膵上縁、肝十二指腸間膜の処理をおこなったのちに、右側からのアプローチによりSMA右側の処理を行う、この時膵頭背側にテープをかけ、hanging maneuverの要領で膵頭部の血流をコントロールしながらSMA右側を切離する。なお、膵実質はScissorsを用い、少しずつ膵実質を離断し、膵管はクリップを行い膵液の漏出を防止している。再建は胃膵吻合で行なっている。RPDに関しては適応の拡大に伴い手術手技を変化させ安全に遂行できていると考える。今後もさらに改変を加え、安全にRPDを遂行できるよう定型化を進めていく

Event date： 2023.5

Venue：名古屋市   Country：Japan  

Event date： 2023.5

Venue：名古屋市   Country：Japan  

Event date： 2023.4

Venue：東京都   Country：Japan  

【背景と目的】ロボット支援下手術，膵癌に対する低侵襲膵切除術の保険収載に伴い，IPMNを対象とした低侵襲膵切除術の数は増加している．一方，全国調査で膵管内乳頭粘液性腫瘍 (IPMN) 切除後再発例 (14.4%)の内，34%は5年以上の経過観察後に再発を認めたことから (Hirono S. et al, J Gastroenterol. 2020)，IPMNに対する腹腔鏡下膵切除の長期予後の検証が必要である．本研究では当院で行ったIPMNに対する開腹手術 (OP)とロボット手術を含む低侵襲膵切除術 (MIP)の長期予後と再発形式を比較検討した．
【患者と方法】2010年1月から2020年11月までに当院でIPMNに対して膵切除術を行った182例 (OP 78例，MIP 104例)を対象とし，臨床病理学的因子，手術成績，再発と全生存期間を２群間で比較した．病変部位 (Ph vs Pbt), 悪性度 (IPMN-LG vs IPMN-HG/invasive IPMN), 主膵管径 (10mm<=)，術式 (OP vs MIP)を変数としてMIPの再発，長期予後への影響を多変量解析で検証した．
【結果】MIPにおいて膵炎の既往が少なく (OP: 28/108, 27%, MIP: 9/78, 11%, P=0.015)，尾側膵切除が多く (OP: 26/104, 25%, MIP: 54/78, 74%, P<0.0001)，出血量は少なかった (OP: 中央値495g, MIP: 136g, P=0.009)．追加切除を要した術中迅速断端陽性例に有意差はなかった (OP: 5/104, 5%, MIP: 1/78, 1.3%, P=0.24)．病理学的にはMIPでIPMN-LGが多かった (OP: 51/78, 65%, MIP: 45/108, 43%, P=0.0021)．予後に関しては，両群間で残膵再発率/局所再発率/遠隔転移・腹膜播種再発率にいずれも有意差はなく，5年再発率/生存率もOP: 17/83%，MIP: 10/95%で有意差はなかった (P=0.1/0.061)．比例ハザードモデルによる予後因子の解析では再発/全生存期間ともに悪性度(HR=31.5/10.7, P=0.0009/0.0024)が有意な因子であり，MIPによる有意な影響は認めなかった (HR=0.84/0.45, P=0.78/0.26)．
【結論】IPMNに対する低侵襲膵切除術は開腹手術との比較では，腫瘍学的に負の影響は認められない．

Event date： 2023.4

Venue：東京都   Country：Japan  

【背景】胃癌治療において抗PD-1抗体といった免疫チェックポイント阻害薬の導入は，これまでの治療に画期的な変化をもたらした．作用機序としてPD-1はCD28とCD226の両方を阻害し，TIGITはCD226を介して腫瘍免疫を阻害する．また，抗PD-1抗体による治療において，免疫染色で評価されるCPSは薬剤投与の指標として期待されている．
【目的】胃癌症例において，T細胞のサブタイプ分類を行い，CPSと相関のある腫瘍免疫関連分子を明らかにする．
【方法】当科で胃切除手術を施行した11症例の胃癌患者から腫瘍部11サンプルを採取し，single cell RNA-sequence解析を行った．その後，T細胞を遺伝子発現の特徴から再分類し，それぞれの遺伝子発現について解析を行った．また，各症例の切除組織を対象に抗PD-L1抗体を用いて免疫染色を行い，CPS分類を行った．CPSは低中高値の３分類に分け，T細胞サブタイプを特徴づける遺伝子発現との関連性を評価した．
【結果】胃癌全11症例の統合解析を行い，CD8陽性T細胞を抽出した．対象細胞数は，10610細胞で，8つのサブクラスターに分類できた．サブグループ別にDEGを評価したところ，CD8陽性T細胞の中でCD226が発現が高いサブグループは，レジデントメモリーT細胞に特徴的であるCD103の発現が高かった．また，CPS高低値でのCD8陽性T細胞におけるPDCD1発現の差は認めなかったが，CPSが50以上の症例はCD8A，CD28，CD226，CD103の遺伝子発現が他の症例と比較して高かった．
【結語】腫瘍免疫におけるCD8陽性T細胞のサブタイプについて特徴的な遺伝子によって分類した．PD-L1が免疫染色上高い発現を認める場合，CD8陽性T細胞にCD28，CD226，CD103発現が高いサブグループを認めていることから，過去の報告から細胞障害性の高いTeffやTem，Trmが関わるHot Tumorであり，抗腫瘍免疫活性が高い可能性がある．

Event date： 2023.4

Venue：東京都   Country：Japan  

Event date： 2023.4

Venue：東京都   Country：Japan  

【背景】HER2陽性胃癌は胃癌全体の約20%を占め、HER2過剰発現は予後不良因子とされている。近年、HER2陽性胃癌に対する免疫チェックポイント阻害薬の有効性が示されており、HER2陽性胃癌における免疫細胞の疲弊化分子はHER2陰性胃癌よりも発現が低いことが示された。しかし、HER2陽性胃癌の腫瘍微小環境に存在する免疫細胞の疲弊化分子の局在については不明な点は未だに多い。
【目的】抗原特異的な細胞障害性機能をもつProgenitor exhausted CD8陽性T細胞 (PE-CD8+T細胞)に着目し、HER2陽性胃癌の治療標的となる疲弊化分子を明らかにする。
【方法】当科で胃切除術を施行した13例の胃癌患者から腫瘍部12サンプルと正常粘膜10サンプルを採取し、single-cell RNA sequencing (scRNA-seq)を行った。その後、免疫染色によりHER2陽性胃癌を同定し、PE-CD8+T細胞における疲弊化分子の発現を比較検討した。
【結果】
胃癌症例12例中、HER2陽性胃癌は2例であった。全22症例の統合解析を行った結果、quality check後のCD8陽性T細胞数は11615であり、6つのクラスターに分類された (Fig. 1)。遺伝子発現の特徴からPE-CD8+T細胞のクラスターを同定し、疲弊化分子の発現を評価したところ、HER2陽性胃癌とHER2陰性胃癌の疲弊化シグネーチャースコア(CTLA4・LAG3・TIGIT・HAVCR2より算出)に有意差を認めなかった (Fig. 2)。さらに、それぞれの疲弊化分子の発現を比較したところ、LAG3はHER2陽性胃癌でHER2陽性胃癌よりも発現が低く、TIGITはHER2陽性胃癌でHER2陰性胃癌よりも発現が高かった (Fig. 3)。
【結論】 HER2陽性胃癌では抗原特異的CD8陽性T細胞におけるTIGITが治療標的となる可能性が示唆された。

Event date： 2023.4

Venue：東京都   Country：Japan  

＜目的＞
ロボット支援下膵切除術(RP)は保険収載後いまだ2年が経過しておらず、多くの施設が導入初期の段階であり浸潤性膵管癌（inv-PDAC）に対しては少ない未だ症例は少ないと思われる。当科では2018年7月よりロボット支援下膵頭十二指腸切除術（RPD）を臨床試験下に開始し、2020年8月からロボット支援下尾側膵切除（RDP）を導入、inv-PDACに対しては2021年からRDP,RPD共に開始している。今回当科での膵癌に対するロボット支援下手術の手術手技の提示および長期成績に関して報告を行う
＜方法＞
2018年7月から2022年6月の間に当科で行ったロボット支援下手術78例（RPD47例
、RDP 31例）を対象とした。inv-PDACに対するロボット支援下手術の適応は術前画像で門脈へ直接接していない症例や他臓器浸潤を認めないものを基本としている
RPD.RDP共にR0切除を重要視した手術を行なっている
＜結果＞
51例中膵癌症例はRPD 19例（inv-PDAC ；12例）、RDP 9例（同；9例）であった。術前診断がinv-PDACである症例に対してはRPD導入後2年8ヶ月後（20例目）に、RDPは導入後5ヶ月後（6例目）に開始された。
手術時間および、出血量をinv-PDAC 群とその他の症例群で比較したところRPDでは（756min; 667min-863min）vs (735min;492min-998min)、（175g;37-476g）vs (300g;17g-1950g)であり、RDPでは（410min; 213min-577min）vs (398min;232min-683min)、（85g;0-440g）vs (258g;0g-1070g)であり手術時間、出血量ともに両群間で有意差は認めなかった。全例R0切除術を達成しており、長期成績に関しては観察期間範囲（3-51ヶ月）において全例無再発生存中である。
＜結語＞
今後の長期観察が必要であるが、inv-PDACに対するロボット支援下手術は安全に導入されていると考えられる。

Event date： 2023.4

Venue：東京都   Country：Japan  

【背景・目的】高齢化社会において偶発的に膵管内乳頭粘液性腫瘍（IPMN）が診断される機会が多くなっている。IPMNは国際診療ガイドラインに沿って手術適応を検討されるが、膵切除術は高侵襲であるため、高齢者に対しては特に慎重な判断が求められる。今回、高齢者におけるIPMN切除例の特徴と膵切除術の安全性について評価した。
【方法】2017年1月から2021年12月までに当科でIPMNに対して膵切除術を施行した75例を対象とし、高齢者群と若年者群で患者背景および周術期短期成績を後方視的に比較検討した。今回の検討では75歳以上を高齢者と定義し、併存膵癌症例は除外した。
【結果】高齢者群は22例、若年者群は53例であった。年齢の中央値は79（75 - 90）歳と68（52 - 74）歳であった。High-risk stigmataを認めた症例はそれぞれ17例（77%）と39例（74%）、術前膵液細胞診で悪性の確定診断を得た症例は8例（36%）と20例（38%）で有意差は認めなかった（P=0.69、P=0.91）。最終病理診断はlow-grade dysplasia: 7例（32%）と16例（30%）、high-grade dysplasia: 8例（36%）と16例（30%）、IPMN with an associated invasive carcinoma: 7例（32%）と21例（40%）であった（P=0.80）。術式は若年者群（26%）よりも高齢者群（45%）で尾側膵切除（DP）が有意に多く（P=0.03）、高齢者群に対して膵全摘は施行されなかった。周術期については、膵頭十二指腸切除術（PD）とDPともに2群間でCD分類IIIa以上の合併症頻度に有意差は認めず（P=0.27、P=0.22）、術後在院日数も同等であった（
P=0.23、P=0.37）。
【結語】高齢者IPMNに対して安全に膵切除術が施行できることが示唆される一方で、高齢者には低侵襲な術式が選択される傾向があった。

Event date： 2023.4

Venue：東京都   Country：Japan  

【背景】腫瘍の縮小・進展において多種多様な免疫細胞から構成される腫瘍免疫微小環境の不均一性が大きく関与していることが知られている。B細胞は液性免疫の中心的な役割を担っているが、近年、「制御性B細胞 (regulatory B cell: Breg)」と呼ばれる免疫抑制的な機能を持つ特殊な細胞集団が存在することが報告されており、抗腫瘍免疫においても重要な働きをもつと考えられている。今回シングルセル解析を用いて食道癌における制御性B細胞に関する解析を行った。
【対象】食道正常粘膜部・扁平上皮癌部の計17検体を対象にシングルセル解析を実行した。シングルセル解析はDrop-seq技術を用いた。
【結果】計17検体から得られたシークエンスデータに対しUMAPを用いた次元圧縮を行った後にクラスター分類を実行しB細胞集団を同定した。B細胞は5,338個検出され、詳細な12のB細胞クラスターへ分類された。各クラスターの発現変動遺伝子解析を行ったがBregクラスターは明確には同定されなかった。BregはIL-10, IL-35などを産生し免疫抑制性の働きを持つことが知られているが、それらを基にgene signatureを作成し各B細胞クラスター間で評価すると各クラスターで発現を認め、特にナイーブB細胞クラスターでの発現が他のクラスターと比較し有意に高値であった。Bregと食道癌の予後との関連を評価するため、食道癌のTCGAデータセットを用いて生存分析を行うと、signature低発現群で有意に無再発生存率が延長していた (p = 0.039)。以上により、Bregは明確な分化サブタイプではなく非常に不均一な細胞集団を表現していることが示唆され、食道癌においてBreg signatureは予後不良因子であり抗腫瘍免疫においては負の働きをしている可能性が考えられた。(
B
【結語】シングルセル解析を用いて微細なBreg関連遺伝子の発現をとらえることができた。今後は術前化学療法の関連に着目しつつ詳細な解析を行う。

Event date： 2023.4

Venue：東京都   Country：Japan  

Event date： 2023.4

Venue：東京都   Country：Japan  

Event date： 2023.4

Venue：東京都   Country：Japan  

腎移植は末期腎不全に対する血液・腹膜透析の代替手段であり、本邦でも生体腎移植が増加している。移植により透析離脱が可能なだけでなく、移植により長期予後の改善も期待できる。腎移植増加による末期腎不全患者の予後改善により腎移植後で免疫抑制剤内服状態の大腸癌患者は今後も増えてくると考えられる。長期間使用する免疫抑制剤による大腸癌の発生・進展に与える影響が懸念される。今回我々は2008年～2022年に当院で大腸癌に対する外科手術を施行した腎移植後大腸癌9例について治療成績と臨床病理学的因子を検討した。男女比は4：5で年齢は60.7 (42-75)歳、腎移植から大腸癌手術までの期間は95.5 (24-180)ヶ月、移植までの透析期間は17.2 (0-36)ヵ月であった。免疫抑制剤は
ミコフェノール酸モフェチルを8例、タクロリムス5例、メトロプレドニゾロン9例、シクロスポリン2例、アザチオプリン1例を使用していた。癌の局在は右側結腸4例、横行結腸3例、直腸2例と右側結腸に多い傾向であった。クローン病に合併する痔瘻癌と高度の腹膜播種を認めた2例は人工肛門造設施行してそれぞれ放射線化学療法、化学療法を施行したが早期にBSCに移行した。残り7例は腹腔鏡で根治手術が可能であった。術後合併症は1例に縫合不全、脳梗塞を発症した。病理組織診断では分化型腺癌が7例、印環細胞を伴う低分化腺癌が1例、扁平上皮癌が1例であった。切除可能であった7例ではT3例、T4例であり、リンパ節転移陽性を1例に認めた。長期予後としては切除不能な2例については非常に予後不良であり、切除可能な症例では4例中1例で早期の肺転移再発を来していた。小数例の検討であり十分な検討ができているとは言い難いが移植後大腸癌は非常にAggressiveなものが多くことが示唆された。

Event date： 2023.4

Venue：東京都   Country：Japan  

Event date： 2023.4

Venue：東京都   Country：Japan  

大腸癌を含む多くの癌において、Tregは他のTcellのはたらきを抑制することで癌の免疫回避に関わっていることが知られている。家族性大腸腺腫症(FAP)患者にはAPC遺伝子の生殖細胞系列遺伝子変異の影響で大腸癌が高頻度に生じ、様々な段階の前癌病変も存在する。FAPは正常部から前癌状態、発癌後の変化を連続的に解析する対象として有用である。また近年、single cell RNA sequence (scRNA-seq)による癌組織のheterogeneity解明が盛んに行われるようになったが、FAPについての解析は上皮細胞中心のものが多く、免疫微小環境については明らかになっていないことが多い。
今回我々は、当科にてFAPに対し手術を行った4人の患者に対し、10X chromium systemを用いてscRNA-seqを施行した。全部で56225個の細胞の遺伝子発現データが得られた。Seurat上で解析を行い、UMAPを作成したところ，26個のクラスターが得られた。FOXP3+ CD4+ TcellをTregとして抽出し、免疫抑制能に関わる遺伝子の発現を3部位で比較した。FOXP3, BATF, IL2RA, TNFRSF18の発現は発癌に伴い徐々に上昇することが明らかになった。
今回の検討で示された結果に基づきFAPの各発癌過程におけるTregの免疫抑制能の変化について考察を加えて報告する。

Event date： 2023.4

Venue：東京都   Country：Japan  

Event date： 2023.4

Venue：函館市   Country：Japan  

Event date： 2023.3

Venue：宮崎市   Country：Japan  

Event date： 2023.3

Venue：宮崎市   Country：Japan  

（はじめに）直腸脱の治療法は多岐にわたるが、まれに直腸腫瘍を合併し治療法に迷うことがある。今回、進行直腸癌を先進部とした直腸脱に対し、Altemeier法に準じた経肛門的手術を施行した１例を経験した。（症例）86歳女性、下血および肛門部腫瘤を主訴に当院を紹介受診した。完全直腸脱および脱出腸管に手拳大の腫瘤を認め、生検で腺癌の診断となった。高齢でPSは3-4、認知症も認めたため侵襲の少ない経会陰的なアプローチを選択し、Altemeier法に準じた切除術およびThiersh法を施行した。切除腸管は14cm、腫瘍径は80×50mmで、病理組織診断はmucin成分を伴う腺癌でpT3(SS)であった。術後経過は良好で合併症や直腸脱の再燃なく経過した。（まとめ）直腸癌を合併した直腸脱に対する経会陰的手術の報告は、本邦において数例のみである。Altermire法に準じた経肛門手術は低侵襲に腫瘍切除を行うことができるため、耐術能に懸念がある高齢者には有用と考える。

Event date： 2023.2

Venue：札幌市   Country：Japan  

Event date： 2023.2

Venue：札幌   Country：Japan  

【背景】HER2陽性胃癌は胃癌全体の約20%を占め、HER2過剰発現は予後不良因子とされている。近年、HER2陽性胃癌に対する免疫チェックポイント阻害薬の有効性が示されているが、HER2陽性胃癌の腫瘍微小環境に存在する免疫細胞について不明な点は未だに多い。
【目的】抗原特異的な細胞障害性機能をもつ疲弊化CD8陽性T細胞に着目し、HER2陽性胃癌の細胞障害性分子の発現を明らかにする。
【方法】当科で胃切除術を施行した13例の胃癌患者から腫瘍部12サンプルと正常粘膜10サンプルを採取し、single-cell RNA sequencing (scRNA-seq)を行った。その後、免疫染色によりHER2陽性胃癌を同定し、疲弊化T細胞における細胞障害性分子の発現を比較検討した。
【結果】
胃癌症例12例中、HER2陽性胃癌は2例であった。全22症例の統合解析を行った結果、quality check後のCD8陽性T細胞数は11615であり、6つのクラスターに分類された。遺伝子発現の特徴からProgenitor exhausted CD8陽性T細胞とTerminal exhausted CD8陽性T細胞のクラスターを同定し、細胞障害性分子の発現を評価したところ、HER2陽性胃癌とHER2陰性胃癌の細胞障害性シグネーチャースコア(GZMB・PRF1・IFNG・TNFより算出)に有意差を認めなかった。さらに、それぞれの細胞障害性分子の発現を比較したところ、IFNGはHER2陽性胃癌でHER2陽性胃癌よりも発現が有意に低く、GZMBとPRF1はHER2陽性胃癌でHER2陰性胃癌よりも発現が有意に高かった。
【結論】 HER2陽性胃癌では抗原特異的CD8陽性T細胞におけるGZMBとPRF1が細胞障害性機能に重要である可能性が示唆された。

Event date： 2023.2

Venue：札幌市   Country：Japan  

Event date： 2023.1

Venue：東京都   Country：Japan  

Event date： 2022.12

Venue：横浜市   Country：Japan  

大部分の大腸癌は環境因子、生活習慣、加齢などにより大腸粘膜に遺伝子変異が蓄積することで発生すると考えられているが全大腸癌の20－30％は血縁者に多発することから家族性大腸癌とも呼ばれている。その中の一部(5％)についてはその原因遺伝子が同定されており遺伝性大腸癌と呼称されている。最近、PD-1阻害薬であるペンブロリズマブがMSI-H進行再発固形癌に保険収載され、今後大腸癌の約2－4％を占めるリンチ症候群が期せずして発見される可能性が高まっている。そこで我々は大腸癌における家族歴の臨床的意義を再検討することとした。1971年3月から2018年12月に当院で手術を施行した初発大腸癌4445例を対象として家族歴の臨床的意義について検討した。家族歴を持つ大腸癌は662例に認め、Propensity score matchingを用いて年齢、性別、腫瘍局在、進行度、腫瘍マーカー、原発巣切除の有無でマッチングして、家族歴を持たない3783例との比較検討を行った。マッチング前の状態で家族歴あり群はなし群と比較すると、若・オ・P<0.001)で、女性が多く(P=0.041)、自覚症状ありが少なく(P<0.001)、局在は直腸に少なく右側結腸に多く(P=0.242)、多発大腸癌は多く(P=0.049、腫瘍マーカーは低く(P=0.266)、遠隔転移はいずれも少なく(P=0.025)、進行度は低い(P=0.143)傾向にあった。そのためそのまま予後を比較すると家族歴なしで有意に予後が悪かった(P=0.025)。両群のマッチングはJMP ver.16を用いてCaliper係数0.01で行った。マッチング後には両群に588名が残り、予後解析を行ったところ予後曲線は完全に重なり予後に差を認めなかった(P=0.943)。家族歴のある大腸癌には様々な臨床病理学的な特徴があるが、家族歴の有無のみで癌の生物学的悪性度に影響を与えるわけではない。

Event date： 2022.12

Venue：名古屋市   Country：Japan  

[Background]
In recent years, per oral endoscopic tumorectomy (POET) has been devised as an operation for benign submucosal tumors of the esophagus. However, large tumor more than 50mm in size is out of the indication. Laparoscopy endoscopy cooperative surgery (LECS) has also been established and well-accepted as an operation for tumors in stomach or duodenum. Herein, we report the case series of thoracoscopy endoscopy cooperative surgery (TECS) and junctional LECS (J-LECS) with POET technique for tumors located in the esophagus or esophago-gastric
junction.
[Cases]
1. A case of leiomyoma with a maximum diameter of 60mm in the lower thoracic esophagus which caused epigastralgia.
2. A case of leiomyoma with a maximum diameter of 70mm on the right wall of the middle thoracic esophagus.
The tumor strongly compressed the trachea, right bronchial artery and azygos vein with a complaint of difficulty of swallowing.
3. A case of 40 mm-sized leiomyoma that was located at the gastric side of esophago-gastric junction compressing the esophagus with a complaint of difficulty of swallowing.
[Surgical procedure]
Surgery was started endoscopically. An entry hole was made about 3cm above the tumor, and a submucosal
tunnel was created to remove the tumor as much as possible. The tumor was dissected almost all around endoscopically. Then, thoracoscopically or laparoscopically, the tumor was resected through a small hole made on adventitial muscularis of esophagus or serosal muscularis of stomach. The defect due to excision was closed with suturing by surgeon, and entry hole was closed with clips by endoscopist. Especially in third case, the defect was very small, and the esophageal mucosa was definitely intact promising the preserved junctional function.
[Postoperative course]
There were no stenosis or leakage in all cases.
[Conclusion]
TECS or J-LECS with POET technique is a very useful minimal invasive procedure.

Event date： 2022.12

Venue：名古屋市   Country：Japan  

転移性膵腫瘍は膵悪性腫瘍の約2%を占め、切除例の原発巣は腎癌、大腸癌、悪性黒色腫、次いで約4%が肉腫と報告されている。間葉性軟骨肉腫は軟骨肉腫の3～10％ を占める稀な腫瘍であり、間葉性軟骨肉腫の膵転移を切除した報告は本邦で2例しかない。今回我々は、間葉性軟骨肉腫の異時性膵転移に対し2度のロボット支援下膵切除術を施行した症例を経験した。
症例は45歳女性。左鼠径部の違和感を主訴に前医を受診した。CTで左股関節部に7cm大の軟部腫瘍と、膵頭部に2cm大の腫瘍を認め精査加療目的で当院紹介となった。CTガイド下針生検で左股関節部の腫瘍は軟骨肉腫が疑われ、切除により間葉性軟骨肉腫と診断された。膵頭部の腫瘍は、超音波内視鏡下穿刺吸引細胞診で軟骨肉腫の転移と診断され、原発巣切除から5か月後にロボット支援下膵頭十二指腸切除術を施行した。その後外来で定期検査を行っていたが、膵切除から7か月後にCTで膵尾部に軟骨肉腫の転移を疑う15mm大の円形腫瘤を認めた。他の部位に転移は認めず、初回膵切除から9か月後にロボット支援下脾合併尾側膵切除を施行した。
間葉性軟骨肉腫の異時性膵転移に対し2度の膵切除術を施行した報告はこれまでなく、文献的考察を踏まえ報告する。

Event date： 2022.12

Venue：名古屋市   Country：Japan  

Persistent descending mesocolon (PDM)は胎生期における固定異常の一種である．S状結腸が内側・右側に変位し，小腸間膜や盲腸周囲に広範囲に癒着していることが多く，広範な癒着剥離が必要となることが多い．今回，当科におけるPDM併存症例を検討し，その特徴と手術成績を解析し，腹腔鏡手術の安全性と対策を検証した．
2010年以降に腹腔鏡下左側結腸・直腸切除術を施行した481例を後方視的に検討した．ロボット支援下手術，側方リンパ節郭清例, 開腹手術既往，他臓器合併切除症例などは除外した．PDMを27例に認めた．PDM併存例は非併存例と比較して出血量が多く(p＜0.01)，手術時間(p=0.01)が延長した．術後在院日数やClavien-Dindo分類III以上の合併症率に差はなかった．結腸間膜短縮により辺縁動脈が下腸間膜動脈に近接する傾向にあり，術中解剖の理解に細心の注意を要した．最近では，可能な血管処理は体腔外で行うようにしている．また直腸後腔の剥離層は通常通り同定が容易で，ここから頭側に剥離層をつなげることも有用である．
PDM併存例に対する腹腔鏡手術は，癒着剥離操作や内側アプローチ剥離層の同定困難などのため出血量が多く手術時間が延長した．しかし術後合併症の発生率は通常と変わらず在院日数の延長もないため，腹腔鏡手術は施行可能と考える．

Event date： 2022.12

Venue：名古屋市   Country：Japan  

＜背景＞
ロボット支援下膵頭十二指腸切除術（RPD）は腹腔鏡に比べて再建で有用でないかと考えられていたが尾側膵切除術（RDP）は腹腔鏡と比べて、その有用性に関して懐疑的な意見も聞かれていた。当科では2016年よりRPDを臨床試験下に導入を開始し、2020年4月の保険収載後にRDPの導入を開始した。今回当科におけるRDPの導入とともに腹腔鏡との違い、その有用性に関してビデオを供覧する。
＜方法と症例＞
当科ではこれまでに38例のロボット支援下膵切除術（膵頭十二指腸切除術；26例、尾側膵切除術;12例）行っている。
RPD導入後の鏡視下膵切除術の内訳は2020年がLDP X例、RDP Y例、2021年がLDPX例、RDP Y例であった。
アプローチ方法に関しては膵上縁から主に剥離を行うSuperior approachを腹腔鏡尾側膵切除術で採用していたが、Robot支援下尾側膵切除術(RDP)の導入に伴い、RDPでも同様のアプローチを採用している。大網を十分に開放後膵上縁操作に移る。総肝動脈(CHA)を確保、taping、そのまま左胃動脈の左側を剥離、左胃膵ヒダを切離する。左胃膵ヒダの背側を十分に剥離、良性疾患では膵後筋膜の背側を、悪性疾患では副腎前面（Gerota筋膜背側）の層を十分に剥離、ガーゼを挿入する。腹腔鏡では可及的に剥離を行っていたが、腹腔鏡よりも可動域制限のないrobotでは、ほぼ膵下縁付近まで剥離を行うことが可能である。その後、膵下縁に沿って切開を行い、標本を摘出する。
＜結語＞
これまでに12例のRDPを施行した。平均手術時間はX分、出血量中央値はYmlであった。RDP導入後のRDPとLDPの比率を鑑みると今後RDPが主体となる可能性が高い。
RDPは比較的導入も平易でありRPDへ向けた準備段階となり、またLDPなしでも行っていく時代が来る可能性があると考える。

Event date： 2022.12

Venue：名古屋市   Country：Japan  

＜はじめに＞
膵癌に対するロボット支援下尾側膵切除術（RDP）は保険収載されて以降、広く普及しつつある。それに伴い、肥満や進行癌など、困難症例を行うことがある。そのような場合、一つではなく様々なアプローチに精通することで対応が可能となる。RDPでは鉗子軸に制限がなく、頭側からのアプローチが有効であることが多い。当科におけるRDPの手術手技を供覧しながら概説する。
＜手術手技＞
総肝動脈を確保後、左胃動脈左側から胃膵ヒダ背側を剥離する。胃膵ヒダを脾上極に向けて切離する。4番アームで切離した胃膵ヒダを足側に牽引すると安定した視野でGerota筋膜および同膜に覆われた左副腎を透見できる。Gerota筋膜を切開し、膵尾側へ向けて膜を意識しながら剥離を続ける。剥離が進むと脾動脈根部を切離することでさらに授動を行うことが可能となり、視野を変えることなく左副腎静脈、左腎静脈前面を露出、さらには膵下縁付近まで剥離可能である。
＜結果＞
2020年8月から2022年4月の間に施行したRDPは27例であった。手術時間中央値、出血量中央値はそれぞれ403分（232-683分）、200ml（0-1070ml）であった。
＜結語＞
頭側からのアプローチは膵癌に対するRDPにおけるアプローチの一つとして有用であると思われる。

Event date： 2022.12

Venue：名古屋市   Country：Japan  

Event date： 2022.12

Venue：名古屋市   Country：Japan  

Event date： 2022.12

Venue：名古屋市   Country：Japan  

潰瘍性大腸炎(UC)の標準術式は大腸全摘である。大腸全摘は剥離範囲が非常に広く、腸間膜の処理、再建など長時間手術が必要になる。低侵襲性からUCに対する大腸全摘にも腹腔鏡手術が導入されつつあるが、さらなる長時間手術が必要となる。経肛門的内視鏡手術は直腸癌手術で確立しつつある方法で、経肛門もしくは会陰的に逆行性に直腸固有筋膜レベルでの剥離を行う。腹腔操作と同時に行うことによって手術時間の短縮が可能になるだけでなく、双方向からの確認の上での剥離が可能となり、安全な手術の一助となる。当科では2020年よりUCに対する大腸全摘術に経肛門内視鏡手術併用を開始した。当科で施行したUCに対する経肛門的内視鏡手術併用大腸全摘術は4症例で男性2例、女性2例で年齢は59.5歳であった。施行術式は全例大腸全摘であり、2例がStapled IAA、1例がHand-sewn IAA、1例が全摘、回腸人工肛門造設であった。手術時間は397.5分であり、出血量は82.5gであった。術後Clavien-Dindo Grade 2の腸閉塞と会陰部
SSIをそれぞれ1例に認めた。2015年からTaTME導入前までのUC大腸全摘症例の手術時間の中央値は546分、出血量は155gであり、経肛門内視鏡手術併用の方が有意に手術時間は短く出血量は少なかった(p?0.05)。本セッションでは当科で施行しているtaTME併用大腸全摘術の手術術式を供覧いただく。

Event date： 2022.12

Venue：名古屋市   Country：Japan  

Event date： 2022.12

Venue：名古屋市   Country：Japan  

癌や腺腫、粘膜下腫瘍などの十二指腸腫瘍は、切除が第一候補となるが、膵頭十二指腸切除は過侵襲となる場合がある。　しかし、EMR, ESDなどの内視鏡的治療は、十二指腸は壁が薄いため遅発性穿孔率が10%弱と非常に高率である。近年、腹腔鏡内視鏡合同手術 (LECS)の低侵襲性と安全性が評価され、十二指腸腫瘍に対する治療として保険収載された。
　当科では、現在まで十二指腸病変17症例（男9：女8、年齢40-79歳）に対してLECSを行った。腺腫8例、上皮内癌3例、NET6例であり、乳頭を基準として口側10例、肛門側7例であった。開腹移行は2例であり、乳頭近傍腫瘍であったため、安全のため開腹移行した症例である。術後合併症としては3例(18%)に発生したが、2例は内視鏡的切除で穿孔がなかったため腹腔鏡での壁補強などの処置を併置せずに終了した症例である。十二指腸縫合補強を行ってLECSを完遂した症例では、全例で合併症はなく術後経過は良好で、術後13日(中央値)で退院していた。17例中10例でESD中穿孔があり、特に乳頭近傍腫瘍であった場合は、十二指腸閉鎖時の乳頭の縫込みに特に注意が必要である。当科におけるデータを供覧するとともに、当科での方針と今後の課題について文献的考察を含めて発表する。

Event date： 2022.12

Venue：名古屋市   Country：Japan  

近年、胃全摘Roux-en Y再建後に食道空腸吻合部が胸腔内に滑脱し、通過障害を誘発するとする報告が散見される。我々は基本方針として、食道空腸吻合は、リニアステープラーを用いたoverlap法でInverted-T型に吻合している。その際、吻合部が腹腔内に引き出せるように食道を十分に剥離しているが、この食道剥離は長期的に見て、胸腔内陰圧による吻合部の胸腔内への滑脱を助長すると考えている。また同様に、挙上空腸間膜の緊張が緩すぎることも滑脱の一因となり得る。これを回避するために、マージナルや空腸動脈を処理せずに犠牲腸管を作成することで挙上空腸に適切な腹腔側への緊張がかかるように配慮している。近年では、犠牲腸管は吻合後に作成しており、これは実際の緊張を見て最適な場所を自在に選ぶことができ、緊張制御に非常に有効である。一方、吻合に際し、食道切離が高位になった場合は積極的に腹臥位による胸腔内再建を選択しているが、その際は空腸動脈を数本切離して長い挙上空腸を作成し、吻合後に仰臥位にて腹側から十分に引き下げて挙上空腸を横隔膜脚に固定している。本工夫をおこなっキ)タ「唇濮甘ΩRY再建210症例中、吻合部の滑脱に関してはこれまで経験していない。同吻合時の工夫に関するビデオと詳細データを供覧して報告する。

Event date： 2022.12

Venue：名古屋市   Country：Japan  

【背景・目的】胃切除術後には胆石の発生リスクが上がることがよく知られている。しかし、その対象手術の多くは胃全摘術や幽門側胃切除術であり、噴門側胃切除術について論じられた報告は少ない。今回、腹腔鏡下噴門側胃切除(LPG)術後における胆石発生リスクについて、後ろ向きに検討した。
【対象】2000年から2017年に当科で施行した胃癌に対するLPG 58例について検討した。
【結果】LPG施行例 58例のうち5例(8.6％)に術後胆石を認めた。また、同時期に手術を施行した他術式の術後胆石発生率は、胃全摘術10.7%、幽門側胃切除術6.7％であった。LPG術後胆石症５例全てが無症候性であり、いずれも無治療経過観察中である。なお、総胆管結石は１例も認めなかった。
【考察・結論】胃切除術後の胆石発生の原因としては、迷走神経切離による胆嚢収縮機能低下、食物通過経路変更によるコレシストキニン分泌低下の可能性などが考えられている。噴門側胃切除術においては、近年の鏡視下手術でも十二指腸に食物が通過する再建形式が多く、胆石発生頻度と内視鏡的胆道アプローチの容易さ、さらには初回胃切除術時に胆嚢付近に手術操作が及ばない点を踏まえると、特に噴門側胃切除術における予防的胆嚢摘出術は不要と考えられた。

Event date： 2022.12

Venue：名古屋市   Country：Japan  

我々は、上腸間膜動脈(SMA)からIMAが起始する分岐異常を有したS状結腸癌に対して腹腔鏡で根治手術を行った症例を経験した。症例は59歳男性。下痢と腹部膨満を主訴に前医を受診しS状結腸癌の診断となった。腹部造影CT検査よりIMAがSMAから分岐することが判明した。切除可能な孤発性の肝転移を有しており、S状結腸癌cT4a(SE)N2M1 cStage?aと診断した。原発巣・転移巣ともに完全切除可能と判断し、まずは原発巣に対して腹腔鏡補助下S状結腸切除術、3群リンパ節郭清を施行した。術中所見ではSMAから分岐したIMAが空腸起始部の背側を通り左側結腸の腸間膜へ流入していた。術前検査では中枢リンパ節に転移は認めなかったため、空腸起始部の腸間膜流入部でIMAを切離し郭清の上限とした。術後は大きな合併症なく経過し自宅退院となった。術後２ヶ月目に肝転移切除術を施行し、術後補助化学療法を行った。現在再発なく術後10ヶ月経過している。IMAの分岐異常は報告が少なく、その大腸癌の至適リンパ節郭清範囲に対して定まキ)タ「辰晋・鬚呂覆ぁIMA分岐異常があった場合には、癌の壁深達度やリンパ節転移の有無に加え左側結腸や直腸の栄養血管を把握し郭清範囲を決定する必要がある。

Event date： 2022.12

Venue：名古屋市   Country：Japan  

Event date： 2022.12

Venue：名古屋市   Country：Japan  

Event date： 2022.12

Venue：名古屋市   Country：Japan  

Event date： 2022.12

Venue：名古屋市   Country：Japan  

Purpose/Background: In Japan, robotic surgery for rectal cancer was covered by health insurance from April 2018. An increasing number of hospitals offer robotic colorectal procedures to the patients. In our institute, we also started robotic surgery for rectal disease from October 2018, since then 55 patients underwent robotic surgery.
Patients and Methods: In this study, we reviewed the medical charts of 55 RSS patients.
Results: Thirteen patients at the start underwent robotic surgery using da Vinci Si system and 42 patients at the end were undergone by da Vinci Xi system. Thirty three patients were male and 22 patients were female, and median age was 63 years old. Tumor location was as above; 11 was in rectosigmoid, 12 was in upper rectum (Ra), 30 was in lower rectum (Rb), 2 was in anal canal (P). Surgical procedure was as above; high anterior resection (n=12), low anterior resection (n=14), super low anterior resection (n=16), intersphincteric resection (n=4) and abdominoperineal resection (n=9). Median surgical time was 401 min. Median blood loss volume was 42.5 ml. No open or laparoscopic conversion was observed. Postoperative complication (Clavien-Dindo?3) was observed in 4 patients.
Conclusion: At the beginning of the introduction, it took a lot of time to set up outside the body and eliminate the arm interference, and the surgery time tended to be long. By changing the model to da Vinci Xi, arm interference outside the body has been resolved. A stable surgical field could be secured in the pelvis, suggesting the feasibility of the robotic surgery. In the future, it is thought that the operation time will be shortened as the robot cases increase. Long-term prognosis is a topic for future study.

Event date： 2022.11

Venue：福岡市   Country：Japan  

直腸癌において局所再発は比較的頻度の高い再発形式である。第一選択は遺残の無い外科的切除だが、局所再発部位により外科的切除はしばしば困難である。また切除可能であっても他臓器合併切除を要する手術は非常に効率の周術期合併症があり、患者に大きな機能障害が残るだけでなく、周術期死亡についても問題となる。本年4月から大腸癌の骨盤内再発に対する重粒子線治療が保険適応となり、その治療成績と比較的低い有害事象に期待が持たれている。しかし重粒子線照射の際はその高い線量分布から隣接臓器との間に距離が必要となり、スペーサー留置が必要になることが多い。今回当科で施行した直腸癌局所再発に対するスペーサー手術について検討した。2013年10月から2022年4月に直腸癌局所再発に対する重粒子線治療の適応でスペーサー手術を施行した10例を対象に検討を行った。正中切開で開腹し腫瘍周囲を剥離、人工スペーサーにより腫瘍表面を被覆・固定した。大網による被覆や骨盤底形成が可能な症例では人工スペーサーは留置しなかった。（結果）男性 6 例、女性 4 例。キ)タネ振冉・陲 63.7 歳（43-72 歳）。 術式はゴアテックスシート留置５例、吸収性組織スペーサー留置1例、大網充填２例、骨盤底形成２例であった。平均手術時間は 249.8分、平均出血量は 278.5g であった。術後在院日数は平均 11.8 日。短期合併症としては、 腸閉塞が１例、水腎症が１例認められた。全症例において重粒子線照射が可能であった。（考察）保険適応に伴い、重粒子線治療症例は増加してくると考えられる。直腸癌局所再発に対する重粒子線治療のためのスペーサー手術は安全で有効であると考えられた。

Event date： 2022.11

Venue：福岡市   Country：Japan  

Event date： 2022.11

Venue：福岡市   Country：Japan  

超高齢者大腸癌に対する腹腔鏡下手術の安全性は明らかでない.今回,80歳以上の高齢者に対する腹腔鏡手術の成績を後方視的に検証した.
2011年から2021年までに手術を行った80歳以上の166例のうち,開腹手術12例を除いた154例を対象とした.Prognostic Nutritional Index, Modified Glasgow Prognostic Score, Controlling Nutritional Statusなどの栄養指標を含めた臨床病理学的因子と合併症率について検討した.
　年齢中央値は84歳(80-95)で男性68例,女性86例.ASA-PSはI:6例,II:119例,III:19例.BMIは21.8(14.8-34),PNIは46.5(24-61.8).腫瘍局在は,右側結腸が79例,左側結腸40例,直腸35例であった. pStage0が3例,I:47例,II:62例,III:33例,IV:9例.手術時間は291.5分(130-725),出血量は35mL(0-2100)であった.開腹移行は他臓器浸潤と腹腔内癒着の2例のみであった．Clavien-Dindo分類(CD)2の合併症を27例(17.5%),CD3以上を16例(10.4%)に認めた.CD4aを1例認めたが手術関連死はなかった.単変量解析で,CD2以上の合併症は手術時間(p<0.01)/出血量(p=0.02)と有意に関連し,栄養指標とは有意な関連はなかった.多変量解析でも手術時間が有意な因子であった(p=0.04).
後方視的検討であり術式選択含めて多くのバイアスは否めないが,高齢者に対する腹腔鏡手術の短期成績は許容できる.しかし手術時間延長や出血量増加は明らかに合併症と関連しており,特に超高齢者では短時間で出血のない手術を心掛けるべきである.

Event date： 2022.11

Venue：福岡市   Country：Japan  

Event date： 2022.11

Venue：倉敷市   Country：Japan  

Event date： 2022.11

Venue：倉敷市   Country：Japan  

Event date： 2022.10

Venue：Fukuoka   Country：Japan  

[Background] Pancreatic ductal adenocarcinoma(PDAC) is one of the most highly malignant tumors which has complex microenvironment. Cancer-associated fibroblasts(CAF) interact directly and indirectly with various immune cells, stromal cells and cancer cells in the tumor microenvironment, which has heterogeneous populations such as myofibroblastic CAF(myCAF), and inflammatory CAF(iCAF). Transformed growth factor-beta 1 activated kinase-1(TAK1) constitutes the cellular hub for several cytokine-mediated signaling, which regulates inflammatory responses such as NF-κB. While, the role of TAK1 in CAFs is not clear. [Purpose]To explore TAK1 functions in CAFs in PDAC microenvironment.[Materials and Methods] CAFs were primary cultured from fresh resected pancreatic cancer specimens. Three-dimensional co-culture models of PDAC tumors and CAFs were made to explore whether iCAFs or myCAFs functions influenced by TAK1. Then we checked the functions of TAK1 positive CAFs using immunofluoresence assay, western blotting (WB) and so on. Next, C57BL/6 mice were subcutaneous injected with organoid mixed with CAFs from KPC mice. [Results]Interference with TAK1 in CAF inhibited tumor cell metastasis and induced myCAF markers increased but iCAF markers decreased, indicating TAK1 inhibitor transform partial iCAF to myCAF. Also TAK1 inhibitors decreased tumor growth and increased immune cell infiltration in the stroma in Vivo.[Conclusions]TAK1 has a possibility to be a therapeutic target focusing on the iCAF which is a conductor of immune system in PDAC.

Event date： 2022.10

Venue：福岡市   Country：Japan  

【背景】
膵solid-pseudopapillary neoplasm（以下SPN）は若年女性に好発する比較的まれな低悪性度腫瘍であり、完全切除により予後良好とされている一方で、再発症例も報告されており適切な術式に関してはいまだ議論の余地がある。
【方法】2008年1月～2022年1月までに当院で手術を施行したSPN40例に関して臨床病理学的評価を行った。
【結果】
全症例の年齢中央値は28歳(15-65歳)で、女性31例、男性9例であった。発見契機としては、腹痛・背部痛17例、他疾患での画像診断検査9例、検診発見14例であった。部位は 膵頭部13例，膵体尾部26例、膵体尾部の多発例が 1 例あった。術式に関しては、膵頭十二指腸切除7例（開腹5例、腹腔鏡1例、ロボット支援下1例）、膵体尾部切除9例（開腹1例、腹腔鏡8例）、脾動脈静脈温存膵体尾部切除1４例（腹腔鏡11例、ロボット支援下3例）、腹腔鏡下Warshaw手術3例、開腹膵分節切除1例、腹腔鏡補助下膵腫瘍核出術6例、を施行した。病理学的所見として、腫瘍径は平均3.2cm(1.0-11.0cm)であり、全例でリンパ節転移や遠隔転移は認めなかった。1 例で術中にrelaced右肝動脈、門脈浸潤が疑われ、肝動脈門脈合併切除・再建を行い、最終病理で門脈浸潤を認めた。1例で術後80.9ヶ月後に他病死(Ｅｗｉｎｇ肉腫の増悪)を認めたが、それ以外の39例においては平均45.2ヶ月キ)(1.3-150.5ヶ月)無再発生存中である。
【結語】
膵SPNは完全切除にて予後は良好である。一方で膵機能温存のために、症例に応じた縮小手術や低侵襲手術の選択が重要である。

Event date： 2022.10

Venue：福岡市   Country：Japan  

【目的】家族性大腸腺腫症(FAP)患者にはAPC遺伝子の生殖細胞系列遺伝子変異の影響で大腸がんが高頻度に生じ，様々な段階の前がん病変も存在する．また, 近年，腫瘍内の不均一性 (heterogeneity)を解析する手法として，Single cell RNA sequence(scRNA-seq)が有用と考えられている．FAPに関連したscRNA-seqの報告は上皮細胞に着目したものが1件のみであり, 免疫細胞に関する報告はない. 本検討ではscRNA-seqによりFAP患者の大腸がん発生の様々な段階におけるCD4+ Tcellを比較することを目的とした．
【方法】当科にてFAPに対し手術を行った4人の患者から，正常部，腺腫部, 癌部の組織を採取し，10X chromium systemを用いてscRNA-seqを施行した．
【成績】56225個の細胞の遺伝子発現データが得られた．Seurat上で解析を行い，UMAPを作成した。既知のマーカー遺伝子発現から、9個の細胞種のクラスターを同定した．CD4+ Tcellのみを抽出して再クラスタリングを行ったところ，7個のクラスターが得られた． 検体採取部位別にCD4+ Tcellの各サブセットの割合を比較したところ, 癌部ではTh1の割合が低いこと, Tregの割合が発がんに伴い上昇すること, Th17の割合が腺腫部で低いこと, Na$(D+Aveの割合が発がんに伴い低下すること, Tfhの割合が発がんに伴い上昇することが明らかになった。また、CD4+ Tcellのexhausted scoreをつけて部位別に比較したところ、exhausted scoreは癌部でのみ上昇していた。
【結論】FAPの各発がん段階における、CD4+ Tcellの違いが明らかになった。

Event date： 2022.10

Venue：Fukuoka   Country：Japan  

背景: 膵癌は予後不良な腫瘍であり、5年生存率が11％未満で致命的な癌の1つである。Acinar-to-ductal metaplasia (ADM)は、膵癌の前癌病変の一時的な状態であり、さらに低悪性度の前癌病変になり、膵癌に進展すると言われている。一方で、血管新生は腫瘍進展における重要なイベントであることが知られている。ただし、CA-ADMにおける血管新生機序に関わっているかは不明である。
目的: 膵癌進展過程におけるCA-ADMや同部の血管新生の意義について、その癌微小環境に着目し検討する。
方法: ヒト膵癌切除組織128例を用いて、CD31、Amylase、CK19、CD68、CD163の免疫組織染色を行なった。KPCマウス膵癌切除36例をCD34、AmylaseとCK19の免疫組織染色を行なった。ヒト膵癌切除組織5例の多重免疫組織染色を行い、CD68、CD163とMMP9の発現と分布を検討し、CA-ADMにおける微小血管密度とtumor-associated macrophage(TAM)の関連を評価した。単球細胞株THP-1をM0とM1、M2型の三種のマクロファージに分化誘導し、in vitroでそれぞれのマクロファージの上清添加条件でのtube formation assayを行った。
結果: ヒトCA-ADMの微小血管密度は膵癌に比べ顕著に高かった。また、CA-ADMについて微小血管密度が高いほど予後不良であった。CA-ADMのCD68、CD163とiNOSの発現量は膵癌に比べ顕著に高かった。TAMのM0型はMMP9を分泌することが明らかになった。M0型上清は、最も血管新生を促進した。一方、KPCマウスCA-ADMの微小血管密度はPDACに比べて高かった。ただし、微小血管密度と生存率が関係なかった。
考察: ヒトCA-ADMの微小血管密度は豊富である。TAMはCA-ADMの血管新生に関与している可能性がある。

Event date： 2022.10

Venue：福岡市   Country：Japan  

【背景】近年、免疫チェックポイント阻害剤であるNivolumabは切除不能進行・再発胃癌に対して標準治療になったが、その奏効率は限定的であり、胃癌免疫微小環境 (TME)の解明は早急に解決すべき課題である。
【対象】当科で胃切除術を施行した胃癌患者13例から採取した腫瘍部12例と正常部10例を対象とし、シングルセルRNAシーケシング (scRNA-seq)を用いて、cancer-associated fibroblast (CAF)のheterogeneityを評価した。
【結論】全22症例の統合解析を行った結果、quality check後のCAF細胞数は2383であり、4の集団に分類された (Fig.1)。4つの細胞集団を遺伝子発現の特徴からinflammatory CAF (iCAF)とmyoblastic CAF (myCAF)の2群に分類した。antigen-presenting CAFはクラスターを形成せず、iCAFとmyCAFクラスターの中に存在していた。機能関連遺伝子の発現を評価したところ、iCAFであるF0クラスターで免疫抑制機能関連遺伝子の発現が最も高値であった。Gene Set Enrichment AnalysisではiCAFはmyCAFと比較して、ケモカイン産生 (FDRq<0.01)や血管新生 (FDRq=0.02)の遺伝子セット発現が有意に高値であった (Fig.2)。
【結論】胃癌CAFにおけるheterogeneityを明らかにし、iCAFは胃癌TMEにおいて免疫抑制環境の形成に関与している可能性が示唆された。

Event date： 2022.10

Venue：福岡市   Country：Japan  

【目的】
癌の進展・縮小に関して腫瘍浸潤リンパ球が重要な役割を担っており、近年では液性免疫の主体であるBリンパ球が固形癌の予後改善に寄与しているという報告が散見される。Ｂリンパ球は抗原刺激に伴って様々な分化サブタイプを示すことが知られているが、腫瘍局所での分化サブタイプのheterogeneityや機能は未だ不明である。本研究では食道癌切除検体を用いたシングルセル解析を行い腫瘍局所でのＢリンパ球サブタイプの詳細な解析を目的とした。
【方法】当科で外科的切除を行った食道正常粘膜部、食道扁平上皮癌部の計17検体を対象に解析を行った。シングルセル解析はDrop-seq systemを使用した。
【成績】計17検体から得られたシークエンスデータをmergeしQuality Checkを施行すると全細胞数：88,954個、総遺伝子数：29,936が同定された。さらにUMAPで次元圧縮を行い、Ｂリンパ球のみを分離し発現遺伝子でクラスター化すると5つのＢリンパ球分化サブタイプ（ナイーブ、活性化、胚中心、メモリー、形質細胞）に分類できた。そのうち形質細胞が最も多く同定され、全Bリンパ球中の53.2％を占めていた。Bリンパ球は様々なタイプの抗体を産生することが知られているが、それぞれの抗体産生Bリンパ球 (IgA, IgD, IgE, IgG, IgM)が腫瘍局所でも明確に分離できた。また、Bリンパ球は抗原提示細胞としての側面も持つが、形質細胞へと分化するとともにMHC class IIの発現が完全に消失しており、分化に伴って抗原提示能を失っていることが分かった。
【結論】Bリンパ球サブタイプの解析を行った。今後、その他の免疫細胞との細胞間相互作用にも注目し、Bリンパ球を中心とした抗腫瘍免疫機能を解明する。

Event date： 2022.10

Venue：福岡市   Country：Japan  

Event date： 2022.10

Venue：福岡市   Country：Japan  

大腸癌はその多くが管状腺癌であり、分化度によって高分化、中分化、低分化に分類されている。浸潤性微小乳頭癌(IMPC)は比較的新しい概念であり、乳癌においては通常型乳癌と比較して悪性度が高く，予後不良と報告されている。大腸においてもリンパ節転移リスクの上昇、幹細胞マーカーの発現上昇や予後不良の報告がある。今回当科において2012年から2016年に切除した原発性大腸癌症例のうち、微小乳頭癌成分をもつ13症例を対象に臨床病理学的因子を解析した。性別は男性6例、女性７例、平均年齢は58.7 (30-69)歳、家族性大腸腺腫症に発生したS状結腸癌の症例を1例に認め、それ以外は通常型の大腸癌であった。腫瘍の局在は盲腸1例、横行結腸3例、下行結腸1例、S状結腸4例、直腸RS2例、直腸2例であり左側結腸・直腸に多い傾向があった。診断時遠隔転移を４例に認め(肝４例、腹膜播種１例)、手術のアプローチは12例(92.3%)に腹腔鏡手術を施行した。リンパ節郭清はD2 4例、D3 13例であった。キ)タッ鶻崚尚椶鯣爾Γ肝磴任錬確磴倭甦釮亡療尚楞・腓砲茲蠎鞜膸爐箸覆辰拭・盈磴聾業・秬攴鋐紊鵬蹴慘屠　・寮攴鋠楾圓靴凸戯独・魄飮・靴討い襦・巴濃・鶻崚尚椶鯒Г瓩覆ぃ肯磴任倭肝禺Ｌ・攴釮・椎修任△辰拭pT分類はT1b 2例、T2 2例、T3 3例、T4a 1例、T4b 1例(膀胱)であった。リンパ節転移は９例中８例に認め、特にT1b 2症例においては３個、４個と比較的高度のリンパ節転移を認め、リンパ節転移をきたしやすい組織型であることが示唆された。８例(88.9％)にL-OHPを含む術後補助化学療法を施行し、3例に再発を認め、5年無再発生存率は66.7％であった。以上よりIMPCを伴う大腸癌ではリンパ節転移が高頻度であり遠隔転移を伴う症例が多かった。今後は遺伝子プロファイルの特徴などについても検討する予定である。

Event date： 2022.10

Venue：福岡市   Country：Japan  

Event date： 2022.10

Venue：福岡市   Country：Japan  

[背景] 頸部食道癌は隣接臓器浸潤やリンパ節浸潤の頻度が高く、進行した状態で発見されることが多い。また、頸部食道癌手術は喉頭合併切除を伴う場合が多いため、根治性だけでなく術後QOLを考慮した治療方針の決定が必要になる。そのため、根治手術か化学放射線療法(CRT)か、また、術前治療は化学療法(NAC)か化学放射線療法(NACRT)か、など検討すべき課題も多い。
[目的] 当科における頸部食道癌症例を後ろ向きに検討し、適切な治療方針を考察する。
[方法] 2014年から2021年までに当科で頸部食道癌と診断された20例について、臨床病理学的因子、治療方針、転帰などについて検討した。
[成績] 頸部食道癌20例のうち、男性13例、女性7例、年齢中央値67歳(48-84歳)であった。表在癌は1例(cStage I)のみで、残りは全て進行癌であった(cStage?:5例、cStage?:8例、cStage ?a:6例)。根治手術を11例(55%)、根治的CRTを8例(40%)に施行し、手術11例のうち8例(73%)で喉頭合併切除を施行し、そのうち2例で長期生存を得た。一方、進行癌に対する放射線治療は10例(根治的CRT：7例、術前CRT：3例)に行い、6例(60%)でCRが得られたが、そのうち根治的CRTを施行した1例で再発（肺転移）を認めた。なお、無再発生存例9例の全例において、初回治療のNACやCRT（根治的CRTも含む）で腫瘍の著明な縮小が得られた(CRまたはPR)。
[結論] 頸部進行食道癌の治療は手術と放射線治療が軸となるが、長期予後を得るためには、初回治療としての術前化学療法あるいは放射線療法が著効することが必要と考えられる。

Event date： 2022.10

Venue：福岡市   Country：Japan  

【はじめに】
新型コロナウイルス感染症(COVID-19)が蔓延している現在、悪性疾患の手術を延期せざるを得ない場合があり、癌患者の診療に少なからず影響を及ぼしている。今回、われわれはCOVID-19療養後に術前検査のSARS-CoV-2 PCRが陽性となり、CTで多発するすりガラス影を認めた進行直腸癌の一例を経験した。【症例】
46歳の女性。便潜血陽性の精査で下部消化管内視鏡検査を施行され、直腸S状部に2型進行癌を認め、手術目的に当科紹介となった。COVID-19罹患後に、保健所の指示で初診日7日前まで隔離されていた。初診時にはCOVID-19関連症状は認めなかった。腫瘍による閉塞症状を認め早期の手術を計画したが、初診日に施行した術前スクリーニングPCRの陽性が判明した。また、CTで左肺上葉に多発するすりガラス影を認めた。予定していた術前精査・手術を一旦延期し、当院の感染対策チーム(ICT)や呼吸器内科に相談した結果、PCRは弱陽性であり発症時期を考えても感染性はないと判断された。2週間後のCTでは左肺上葉のすりガラス影は消失しており、COVID-19関連の陰影と考えられた。初診日から25日後(感染から約6週間)に腹腔鏡下直腸高位前方切除術を施行し、術後合併症なく退院した。関わった医療関係者にCOVID-19の発症を認めなかった。
【まとめ】
COVID-19罹患後の予定手術時期については、手術の緊急性、感染性に加えて術後合併症リスクの観点から判断される。日本麻酔科学会は、待機可能な手術は感染診断から7週間以降に予定することを推奨している。本症例は腫瘍による閉塞症状を認めており、できるだけ早期の手術が必要であった。進行癌患者では手術まで7週間の猶予がない場合もあり、院内ICTや麻酔科医との連携・相談は重要である。さらには胸部陰影を認めた場合は短期間でのCT再検も考慮すべきである

Event date： 2022.10

Venue：Fukuoka   Country：Japan  

Event date： 2022.10

Venue：神戸市   Country：Japan  

There is little evidence of adjuvant chemotherapy for elderly patients since they often exclude from the clinical study of chemotherapy. We examined the clinicopathological factors for elderly patients with Stage III colorectal cancer over 70 years old. Ninety-seven patients were enrolled in this study between 2007 and 2016. The study included 45 male and 53 female with a median age of 77.1 years (range=70-92 years). Tumor was located in right side colon (n=45), left side colon (n=34) and rectum (n=18). Fraility was found in 14 patients (ASA 3). Morbidity (Clavien -Dindo ?2) were observed in 20 patients. Pathologically, high histological grade (mucinous or poorly differentiated adenocarcinoma) was observed in 10 patients. In addition, T4 and N2 were observed in 14 and 16 patients, respectively. Adjuvant chemotherapy was performed in 62 patients including 15 L-OHP-contained regimens. In the patients with PS 3 or more, there is no indication
for adjuvant chemotherapy regardless patients’ age in our department. In the patients with PS 2, we determined the indication for chemotherapy based on cancer stage and general condition. In 35 patients without adjuvant chemotherapy, the cause of omitting chemotherapy was poor PS (n=18), patients preference (n=14). Subgroup survival analysis showed that omitting adjuvant chemotherapy was significant risk factor for cancer recurrence in T4 and N2 group (p=0.0293) though there was no statistical difference whether omitting adjuvant chemotherapy or not in non T4 and N2 group. Although this study is retrospective setting and there are many biases, adjuvant chemotherapy should be performed for the good PS elderly patients with locally advanced or many lymph nodes metastasis.

Event date： 2022.10

Venue：神戸市   Country：Japan  

Purpose: Tumor infiltrating lymphocytes play an important role in the tumor progression. Recently, several reports showed that B lymphocytes, the main components of humoral immunity, contribute to improve prognosis in tumors. B lymphocytes are known to differentiate into various subtypes with antigen stimulation. However, the heterogeneity of tumor infiltrating B lymphocyte subtypes is not clear. Here we examined the heterogeneity of functions and subtypes of TIL-Bs in esophageal squamous cell carcinoma using single-cell transcriptome analysis.
Experimental Design: We examined 17 specimens including 7 normal tissues and 10 tumor tissues which were resected surgically at our department using single-cell transcriptome with Drop-seq system.
Results: We identified 88,954 cells and 29,936 genes with analysis of the sequence data. Furthermore, B lymphocytes were extracted and there were 5,338 cells of B lymphocytes, including 1,982 cells of normal tissues and 3,376 cells of tumor tissues. We classified 5 subtypes of B lymphocytes according to the canonical marker genes. Of these subtypes, antibody-secreting cells (ASCs) were most frequently and accounted for 53.2% of all B lymphocytes. Evaluating the functional genes, the functions of antigen-presentation and co-stimulation were completely lost during differentiating into ASCs. Focusing on germinal-center B cells (GCB, n=114), which are known to play a central role in germinal center reactions, we evaluated the gene set consisted with co-stimulation factors and the expression of the gene set in GCBs of tumor were elevated significantly (p=0.028). It suggested that GCBs in tumor may induce the germinal center reaction actively at the tumor site.
Conclusion: We analyzed the heterogeneity of TIL-Bs. We will focus on cell-interactions with other immune cells in anti-tumor immunity.

Event date： 2022.10

Venue：千葉市   Country：Japan  

Event date： 2022.10

Venue：千葉市   Country：Japan  

【はじめに】
多発大腸癌に対しては，各々の腫瘍部位に応じてCMEを意識したリンパ節郭清を行う必要がある．右・左側結腸に離れて存在する場合や異時性大腸癌の場合は，温存結腸の血行を考慮して郭清しなければならない．今回当科における多発大腸癌に対するCMEを意識した手術戦略を提示し，手術成績を後方視的に検討する．
【対象と方法】
2012年から2021年に手術を施行した大腸癌症例の中で，同時性・異時性多発癌を対象とした．根治的内視鏡切除を施行した腫瘍は除外した．患者背景，腫瘍局在と術式，術後合併症などを後方視的に検討した．異時癌では初回手術記録を入手し，可能な限り造影CTで血管解剖を把握した．2018年以降はICG蛍光法による腸管血流評価を導入している．
【結果】
58例に異時性・同時性大腸癌に対して手術を施行した．3例のみ開腹手術が行われ，その他は全例腹腔鏡手術で開腹移行例はなかった．同時性多発症例は45例で，13例は吻合部が2か所，30例は1か所，2例は永久人工肛門造設術であった．吻合部が2か所の群はそれ以外と比べて手術時間が長くなる傾向にあった（446.4 vs 355.2分, p=0.11）．異時癌13例のうち2例は開腹術後の開腹術，4例は開腹術後の腹腔鏡手術，7例は初回・再手術ともに腹腔鏡手術を施行された．Clavien-Dindo分類III以上の合併症を同時性6例（IIIbイレウス1；IIIa［骨盤死腔炎2；縫合不全2；イレウス1］)・異時性2例（IIIb胆嚢穿孔；IIIaイレウス）に認めた．縫合不全症例は共に直腸癌で，ICG血流評価未施行症例であった．4例に再発を認め，肺転移2例，肝転移1例，1例は異時癌症例の播種再発であったが，初回の進行癌の再発と考えられた．不十分な郭清によると思われる局所再発症例はなかった．
【まとめ】
再発例や縫合不全率からみて，当院における多発癌に対する術式選択は妥当と考えられた．縫合不全2例のうち1例は右結腸切除+ISRの症例で血流障害が疑われた．ICG血流評価導入後は縫合不全の発生はなく，異時癌や2か所吻合が必要な手術では必須と考える．腹腔鏡下手術は開腹術後の異時癌に対しても可能と考えるが，1例に術後胆嚢穿孔症例を認めており，適応は慎重に行うべきである．

Event date： 2022.10

Venue：千葉市   Country：Japan  

憩室炎は時に増悪により膀胱をはじめとする他臓器への瘻孔を合併することがあり、
狭窄待機的とともに外科手術の適応となる。腹腔内の炎症や癒着などを伴う複雑性憩
室炎に対する鏡視下手術の安全性及び妥当性はいまだ明らかでない。本研究の目的は
他臓器との瘻孔を伴う憩室炎に対する腹腔鏡手術安全性及び妥当性を検証することと
術中の工夫について供覧することである。当科にて2012年から2019年に鏡視下手術を
施行した他臓器に瘻孔を伴う憩室炎17症例を対象とした。男性9例女性8例、年齢61.6
(44-78)歳、症状は便性帯下2例、腹痛6例、糞尿/気尿8例、排尿時痛4例、発熱5例(重
複含む)、罹患範囲はS状結腸10例、下行~S状結腸6例、全結腸1例、瘻孔形成臓器は膀
胱12例、膀胱及び小腸1例、小腸2例、膣断端2例であった。膀胱瘻のみでの検討では
術前検査の検査精度はそれぞれ瘻孔診断能CT 8/13 (61.5%)、消化管造影3/10 (30.0%
)、MRI 6/6 (100%)とMRIが瘻孔診断能に優れていた。瘻孔の開口部が尿管に近接して
いる症例には全身麻酔導入後に尿管ステントを挿入した(10/13例)。16症例中結腸膀
胱瘻の2症例において腹腔内の著明な炎症癒着により開腹に移行した。人工肛門は回
腸3例、Hartmann２例造設したが全例で閉鎖可能であった。結腸膀胱瘻に対する瘻孔
部処置は多くがトリミング後に2層に縫合閉鎖を行っていたが、リークテストでも瘻
孔からの色素流出を認めない2例では1層の補強縫合にとどめた。結腸膣瘻の手術症例
は2症例みとめたがどちらも婦人科で子宮全摘出後であり、膣断端との瘻孔であった
。術後合併症としては2症例腹腔内膿瘍を認めたが、縫合不全は認めず、両者ともに
保存的に軽快した。当科における瘻孔を有する結腸憩室炎に対する鏡視下手術は短期
成績からは安全で妥当なものと思われた。鏡視下手術における術中の工夫としては腹
腔鏡の下からのぞき込むように見ることができる視野を利用して瘻孔部の周囲、特に
背側や奥の剥離を先行して出来るだけ授動を行った後に瘻孔部を切開することで瘻孔
を形成した臓器の損傷を最小限にできると思われる。当科における術中の工夫につい
て手術ビデオを供覧しながら概説する。

Event date： 2022.10

Venue：長崎市   Country：Japan  

腸回転異常症は,胎生期に腸管の回転・固定異常を生じる稀な疾患で,多くは乳幼児期に診断される.今回,腸回転異常を伴う上行結腸癌に対し腹腔鏡下切除術を施行した1例を経験した.
68歳男性.貧血の精査で上行結腸に全周性2型進行癌を認め,手術目的に紹介となった.造影CTで単発肝転移と多発肺転移を認めた.全小腸は右側に位置し,Treitz靭帯の形成不全が疑われた.上腸間膜動脈は同静脈(SMV)の背側に位置していたが,SMV rotation signはなくnonrotation typeの腸回転異常と診断した.腹腔鏡下右結腸切除術を施行.右側結腸は内側の固定をほとんど認めず,空腸は十二指腸下行脚からそのまま右側に位置した.回盲部からの後腹膜アプローチで授動を開始し,横行結腸頭側から肝弯曲部授動を完了.最後に回結腸動脈(ICA)および右結腸動脈を同定して根部で処理,D3郭清を行った.手術時間は185分,出血は少量.その他腸管同士の癒着などはなかった.術前のCT通りICAは通常より末梢で分岐しており,解剖の把握に造影CTが非常に有用であった.
腸回転異常症は時に腹腔内の高度癒着を伴い,特にmalrotation typeでは血管走行変化も生じやすい.術前CTで十分なシミュレーションを行い正確な解剖を把握することで,安全に腹腔鏡下手術を施行できる

Event date： 2022.10

Venue：長崎市   Country：Japan  

潰瘍性大腸炎(UC)においては近年薬物療法の劇的な進歩によりUCの炎症がコントロー
ルされた結果,Colitic cancerなどの腫瘍性病変の手術割合が増加している.腫瘍性病
変を伴うUC症例について検討し,臨床病理学的所見を比較検討した.2002年1月~2021年
6月のUC121例のうち腫瘍性病変を伴う49例を対象に検討を行った。平均年齢は55.9(2
5-81)歳でUCと診断されてからの罹患期間は16.5(0-47)年とUC全体より明らかに高齢
で罹患期間が長かった(P＞0.01).組織学的にはHigh grade dysplasiaのみを11例,管
状腺腫を3例に認め,35例が癌と診断されていた.癌周囲にDysplasiaを伴う症例を23例
,癌が低分化または粘液癌を呈する症例を4例に認め,Dysplasiaを伴わない症例を8例
に認めた.癌合併症例のうちDysplasia/低分化/粘液癌合併大腸癌症例(A群)とそれ以
外の大腸癌症例(B群)を検討するとA群の方が若年(55.9 vs 62.1才),罹患期間が長く(
19.4 vs12.8年),進行症例が多い傾向があり,B群は通常型大腸癌の特徴と類似した結
果であった,またB群では背景腸管の炎症が軽度な傾向があり、粘膜の組織学的治癒の
症例も散見された.結語：UC合併大腸癌にはColitic cancerの特徴を持たない病変が
あり,これらは通常型大腸癌に類似した所見を持っていた.

Event date： 2022.9

Venue：東京都   Country：Japan  

本邦における食道癌のうち85%以上が食道扁平上皮癌(ESCC)である。Stage II, IIIに対しては術前化学療法(NAC)後の外科的切除が標準治療とされているが、その5年生存率はそれぞれ56%, 29%と満足のいくものではない。近年、免疫チェックポイント阻害薬の開発、適応が目覚ましく、その普及に伴い免疫学的側面から腫瘍微小環境を解明する試みも全世界的に加速しており、その手段として単一細胞レベルのmRNAを網羅的に解析するsingle cell RNA-sequence(scRNA-seq)の技術が盛んに用いられるようになってきている。本研究では、腫瘍微小環境の一端を担う腫瘍浸潤T細胞(TIL)をscRNA-seqを用いて解析し、NACの有無で比較検討することで、NACがTILにもたらす影響を解明することを目的とした。
当科においてESCCに対し手術、生検を行った症例より正常食道粘膜および癌部を採取しscRNA-seqを行い、同定されたT細胞集団を細分化して、それぞれの機能に着目して解析を行った。
正常部12症例、癌部18症例の統合解析を?ってCD45+細胞を抽出したところ、quality check後の細胞数は83,151で、11のクラスターに分割された。既知の特徴的な遺伝子の発現パターンからそれぞれのクラスターが対応するcell typeを同定し、その中からT細胞を抽出した。CD4+ T細胞(CD4T, n=6,453)は6つにクラスタリングされ、差次的発現遺伝子から既知のsubtypeへ分類することが可能であった。正常部に比べ癌部でregulatory T cellの割合が多く、いずれのクラスターにおいても疲弊化に関する遺伝子発現は癌部において有意に高値であった。癌部においてgene setを用いてNACの有無で比較したところ、NAC施行群では非施行群と比較しtype1 helper CD4Tのmemory化が促進され、exhaust CD4Tの疲弊化が妨げられていることが示唆された。
また、CD8+ T細胞(n=8,005)も6つにクラスタリングされ、こちらも差次的発現遺伝子から既知のsubtypeを同定した。いずれのクラスターにおいても疲弊化遺伝子は癌部で有意に高発現しており、gene setを用いた癌部の評価では、NAC非施行群と比較し施行群でapoptosisが回避され、memory化が促進され、疲弊化が妨げられていることが示唆された。
以上から、NACがESCCのTILの機能的なheterogeneityに変化をもたらしている可能性が示唆された。

Event date： 2022.9

Venue：東京都   Country：Japan  

Event date： 2022.9

Venue：東京都   Country：Japan  

Event date： 2022.9

Venue：東京都   Country：Japan  

[背景] 頸部食道癌は食道癌全体の約4-5％と比較的発生頻度が少ない疾患である。また、進行癌で発見されることが多いうえ、隣接臓器に浸潤しやすく、手術においては喉頭合併切除を伴う場合が多い。また、頸胸境界部領域に癌が及ぶ場合、頸部食道切除のみで良いか、胸部食道まで切除すべきか（食道全摘・亜全摘）、同様にリンパ節郭清も含めた切除範囲についても議論がなされている。さらに、喉頭合併切除が避けられない場合においても術前治療（化学療法(NAC)や化学放射線療法(CRT)）を行うべきか、など検討すべき課題も多い。
[目的] 当科における頸部食道癌手術症例を後ろ向きに検討し、適切な治療方針を考察する。
[方法] 2014年から2021年までに頸部食道癌手術を行った14例について、臨床病理学的因子、治療方針、転帰などについて検討した。
[成績] 頸部食道癌手術14例の内訳は、男性10例、女性4例、年齢中央値68歳(56-71歳)であった。全て進行癌であり、cStage II: 5例、cStage III: 4例、cStage IVa: 5例であった。手術は根治手術12例、根治的CRT後の救済手術1例、CRT後の瘢痕狭窄解除目的での手術１例であった。根治手術12例の術式は、喉頭温存手術4例（33%：頸部食道切除２例、食道亜全摘2例）、喉頭合併切除8例（67%：頸部食道切除４例、食道全摘4例）であった。また、術前治療は12例中10例に行われた（NAC：7例、CRT：３例）。そのうち、喉頭温存手術例では4例全てに術前治療を施行し（NAC：２例、CRT：2例）、術前CRTを施行した２例はいずれも完全寛解(CR、Grade 3、pStage 0)であった。また、再発は12例中5例にみられたが、このうち頸部リンパ節のみに再発した１例は、その後の治療に奏効して長期生存が得られた。なお、食道切除範囲・郭清範囲の違いによる再発率の差は・オ桷C瓩蕕譴覆・辰拭俸・・・酸攴釗6例中3例再発、食道全摘：4例中2例再発）。
[結論] 頸部食道進行癌に対しては病変の広がりに応じた術式・切除範囲を選択すればよいが、特に喉頭温存をめざす場合には、術前化学放射線療法が有効である可能性がある。

Event date： 2022.7

Venue：横浜市   Country：Japan  

＜目的＞
ロボット支援下膵切除術(RP)は保険収載後いまだ2年が経過しておらず、多くの施設が導入初期の段階であり良性～低悪性度腫瘍が適応とされるべきである。膵癌に関してはIPMC,non invasive症例は導入初期から適応とされている施設もあるが、浸潤性膵管癌（IDC）は随伴性膵炎を伴い、難易度が高く、その適用は慎重に開始すべきである。当科では2018年7月よりロボット支援下膵頭十二指腸切除術（RPD）を臨床試験下に開始し、2020年8月からロボット支援下尾側膵切除（RDP）を導入、膵癌に対しては2021年からRDP,RPD共に開始している。今回当科での膵癌に対するRPD,RDP導入までの道のりおよびその成績を報告する。
＜方法＞
2018年7月から2021年11月の間に当科で行ったロボット支援下手術51例（RPD33例、RDP 19例）を対象とした。膵癌に関しては術前画像で門脈へ直接接していない症例や多臓器浸潤を認めないものはRDP,RPDが可能かを症例個々で検討している。
＜結果＞
51例中膵癌症例はRPD15例（inv-PDAC；4例）、RDP4例（同；4例）であった。術前診断がIDCである症例に対してはRPDが導入後2年8ヶ月後（20例目）に、RDPは導入後5ヶ月後（6例目）に開始された。なお、いずれも術前診断がT3N0M0 stage IIAの症例であった。膵癌症例は全例R0切除であり、周術期死亡は認めていない。
手術時間および、出血量をIDC群とその他の症例群で比較したところRPDでは（820min; 683min-863min）vs (739min;522min-996min)、（185g;37-450g）vs (265g;17g-1950g)であり、RDPでは（444min; 338min-467min）vs (413min;232min-683min)、（112g;0-344g）vs (258g;0g-1070g)であり手術時間、出血量ともに両群間で有意差は認めなかった。長期成績に関しては観察期間範囲（2-24ヶ月）において全例無再発生存中である。
＜結語＞
膵癌へのロボット支援下手術はチームとして2年以上が経過したのちに導入を行っている。短期的、長期的経過からみても安全に導入ができていると考えられる。

Event date： 2022.7

Venue：横浜市   Country：Japan  

【背景と目的】
血液中の腫瘍マーカー（CEA、CA19-9、CA-125）は癌患者の治療効果判定を評価するためのマーカーとして使用されているが，膵癌患者の早期診断においては有用とはされていない。今回SNP解析によるgenotypingを用いて患者をグループ化し、グループごとに新たなカットオフ値を設定することにより、切除可能膵癌の早期診断精度を改善することを目的とした。
【方法】
2002年から2018年に膵癌ハイリスク患者の前向きスクリーニングにおいて、膵癌を発症しなかった504人の患者（コントロール群）の血液サンプルをELISAを用いて腫瘍マーカー(CA19-9，CEA，CA-125)値を測定した。またコントロール群において腫瘍マーカーに影響する遺伝子（FUT3、FUT2、ABO、GAL3ST2）のSNP解析を行い、SNPによるグループごとの腫瘍マーカーのカットオフ値（特異度を99%に設定）を決定した。これらは診断精度の検証のためTrainingセット、Validationセットにて検証を行なった。さらにコントロール群のグループ毎のカットオフ値を2010年から2017年に膵癌治癒切除を受けた245人の膵癌患者に当てはめることにより、SNP解析を用いたgenotypingによる切除可能膵癌の診断精度の評価を行った。
【結果】
CA19-9に関して、SNP解析を用いてコントロール群を新たに４つのグループ分類できることを発見し、それぞれのグループのカットオフ値を特異度を99%と設定することにより以下のように決定した。（A群：9.5U/mL, B群：23.5U/mL, C群：38.1U/mL, D群：66.6U/mL）
上記カットオフ値を用いて膵癌切除症例を評価したところ、SNP解析によりグループ分けをすることにより診断精度は特異度:99%にて感度は60.8%と改善した（非分類群の感度；52.7%）
CA19-9を発現しないLewis抗原陰性患者（FUT3 homozygous mutation)を除いた場合、SNP解析を用いることにより特異度:99.3%にて感度は66.4%と診断精度は良好であった。
SNP解析を用いたCA19-9、CEA、CA125のコンビネーションにより、感度:66.1%、特異度:95.4%と切除可能膵癌における診断精度の改善を認めた。
【結論】
腫瘍マーカー（CA19-9、CEA、CA125）の発現に関わるSNP解析を用いて新たなカットオフ値を設定し、患者ごとの個別化診断を行うことにより切除可能膵癌の診断精度を改善することができる。

Event date： 2022.7

Venue：横浜市   Country：Japan  

【はじめに】近年,腹腔鏡下結腸切除術における体腔内吻合(IA)の有用性が報告されるようになり,導入する施設も増えている.当院でも2020年9月にIAを導入し,慎重な症例選択のもと行っている.今回,当院における体腔内吻合手術手技を供覧し,その短期成績を報告する.
【対象と方法】2020年9月にIAを導入し,計18例にIAを施行した.患者背景,腫瘍局在と術式,在院日数,合併症率などを後方視的に検討した.対象群は2016年4月以降の体腔外吻合(EA)例(n=143)とし(除外:DST例,非吻合例,他臓器合併切除例),その短期成績を比較した.IA例では,基本的に機械的と化学的前処置(カナマイシン,メトロニダゾール)を施行.結腸小腸吻合は機能的端々吻合法(FEE)で,結腸結腸はFEEまたはOverlap法で行っている.基本的にPfannenstiel切開創から標本を摘出する.
【結果】IA群18例のうちFEEを12例(右側結腸切除[RC]8例,横行結腸[TC]2例,左側結腸[LC]2例),Overlap法5例(TC3例,LC2例),Delta法を1例(TC)に施行した.IAとEA群で年齢,性別に差はなかった.BMIはIA/EA群24.1/22.2とIA群でやや高く(p=0.06),ASA-PSはIA群でPS1患者が多かった(p<0.01).腫瘍径やpStageに差はなかったが,T因子はIA群でTis/1/2症例が多い傾向にあった(p=0.08).手術時間はIA群で有意に長かった(289.5vs263分,p=0.02)が,出血量,術後在院日数,合併症率には有意差を認めなかった.IA群の2例にSSIを認めたが(11.1%,,縫合不全と蜂窩織炎),EA群(9.8%)と有意差は認めなかった(p=0.69).体腔内吻合時間(追加補強の縫合時間含む)の中央値は43.5(26-60)分であった.間膜処理時間は35.5(14-71)分で,間膜処理からの合計時間がEA群に比べキ)タ「深蟒兒・屬留篦垢陵廾・塙佑┐.一方,RCにおける器械吻合例のみに絞って解析すると(IA群8例,EA群96例),手術時間にも有意差を認めなかった.
【まとめ】IAは導入から間もなく手技がやや煩雑で手術時間が延長するが,その他の短期成績は許容しうる.特に結腸結腸吻合手技の定型化や術者の経験の積み重ねにより,時間短縮は可能と考える.2例にSSIを認めており,さらなる汚染対策が必要である.現時点で,腹腔内脂肪の少ない痩せ型患者におけるIAの利点については議論の余地がある.今後は播種再発のリスクやPfannenstiel切開の恩恵の評価など長期成績・予後を含めた検討が必要である.

Event date： 2022.7

Venue：横浜市   Country：Japan  

【背景】胃癌は世界で減少傾向であるものの本邦における罹患率・死亡数は依然として高い。近年、免疫チェックポイント阻害剤であるNivolumabは切除不能進行・再発胃癌の3次治療として推奨されており、胃癌における腫瘍免疫微小環境 (TIME) についての報告は増加している。しかし、免疫チェックポイント阻害薬の奏効率は限定的であり、胃癌TIMEに存在する免疫細胞については不明な点が未だに多い。
【対象】当科で胃切除術を施行した13例から採取した腫瘍部12例と正常部10例を対象とした。
【結論】全22症例の統合解析を行った結果、quality check後のCD8陽性T細胞数は11615であり、6の集団に分類された。細胞障害性機能が高い3つの細胞集団を遺伝子発現の特徴からterminal exhausted CD8陽性T細胞、progenitor exhausted CD8陽性T細胞、acute activated CD8陽性T細胞と判断した。これらの3つの細胞集団はすべて、正常部と比較して胃癌で有意に殺細胞性の上昇 (p<0.001)と疲弊化 (p<0.001)の上昇を認めた。また、これらの細胞集団は正常部と比較してG2M期に存在する細胞の割合が増加していた。さらに、疑似時間軸でterminal exhausted CD8陽性T細胞とprogenitor exhausted CD8陽性T細胞はacute activated CD8陽性T細胞とは異なる分化の軌跡をたどる可能性が示唆された。
【結論】胃癌ではterminal exhausted CD8陽性T細胞とprogenitor exhausted CD8陽性T細胞が抗腫瘍免疫において重要な役割を果たしており、CD8陽性T細胞の中でもこの2つの細胞集団を標的とした免疫療法が有用である可能性が示唆された。

Event date： 2022.7

Venue：横浜市   Country：Japan  

【背景】腫瘍局所の微小環境には、T細胞、B細胞、樹状細胞など多種多様な免疫細胞が存在し腫瘍の進展・縮小に大きく関与している。また近年、切除不能食道癌に対し抗PD-1抗体が保険適応になるなど、消化器癌における免疫微小環境の理解は急務である。Ｂ細胞は液性免疫における中心的役割を担っているが、腫瘍局所で実際にどのような機能を発揮しているのかは未だ不明である。今回、食道扁平上皮癌の切除検体を用いてシングルセル解析を行い、腫瘍内B細胞の詳細な機能解析を行った。
【対象】食道正常粘膜部・食道扁平上皮癌部の計17検体を対象にシングル解析を行った。シングルセル解析はDrop-seq技術を基盤とした10x genomics社のChromiumコントローラーを用いた。
【結果】計17検体から得られたシークエンスデータを一旦すべてmergeした上で、Rソフト上のSeuratパッケージを用いてQuality Checkを施行すると、全細胞数：88,954個、総遺伝子数：29,936が検出された。それらの検出細胞に対してUMAPを用いて次元圧縮を行いクラスター化を実行したところ、25のクラスターに分けられた。各クラスターの発現遺伝子からB細胞を分離し再クラスター化を実行すると、新たに5つのクラスターに分けられた。B細胞全体で5,338個検出され、正常部：1,982個、腫瘍部：3,376個が同定された。それぞれのクラスターに対し、代表的な機能遺伝子発現の解析を行った。B細胞の活性化を示すNF-κB経路関連遺伝子は活性化B細胞クラスターで強く発現していた。一方、細胞増殖シグナルであるMYC経路関連遺伝子は活性化B細胞・メモリーB細胞クラスターで発現していた。また、PD-1, PD-L1/2の発現にもクラスター間で不均一性がみられた。以上の結果より、B細胞クラスター間で活性や細胞増殖シグナルの不均一性ｋオ梟タ減澆垢襪海箸・・・辰拭
【結語】B細胞の各サブタイプにおける細胞活性の評価を行うことができた。今後は術前化学療法の有無や正常部・腫瘍部での比較を行い、より詳細な免疫微小環境の解析を行っていく。

Event date： 2022.7

Venue：横浜市   Country：Japan  

Event date： 2022.7

Venue：横浜市   Country：Japan  

Event date： 2022.7

Venue：横浜市   Country：Japan  

Event date： 2022.7

Venue：横浜市   Country：Japan  

＜背景と目的＞
我が国の食道癌根治手術後再発は28-47%と高率に認められる。また、再発診断時からの生存期間中央値は5-10か月であり、再発後の転帰は不良である。しかし、中には再発後の治療により長期間病勢を制御できたものや、完全奏功して長期生存できた例もみられる。そこで、食道癌再発後に長期生存できた症例の臨床病理学的特徴について検討した。
＜対象と方法＞
2000年から2016年までに当科で施行した食道癌根治手術288例のうち、術後再発を認めた81例(28.1%)について検討した。術後再発までの期間(中央値)は11.1か月、再発からの生存期間中央値は12.1か月であった。
＜結果＞
再発例81例のうち、再発後5年以上の長期生存例は3例(3.7%)であった。
再発後の生存期間が長い群(2年以上：L群）は、生存期間が短い群（6か月以内：S群）よりも、手術から再発までの期間が長かった（L群15.2か月、S群9.8か月、p=0.022）。また、手術時の進行度や再発形式による差はみられなかった。なお、5年以上長期生存例3例の再発後の治療内訳は、2例に化学放射線治療を施行、1例に化学療法を施行した。また、1例はニボルマブが有効であった。
＜結論＞
食道癌術後再発までの期間が長いほど、再発後の生存期間も長くなる傾向がある。

Event date： 2022.7

Venue：横浜市   Country：Japan  

【はじめに】
膵臓癌は予後不良な疾患であり、その原因の一つとして標準治療である化学療法に対しても抵抗性があることが挙げられる。化学療法抵抗性の機序に関して、膵癌においては高度な不均一性があり、さらに周囲の間質との相互作用も考慮に入れる必要もあることから、まだ十分に解明されていない。
近年シングルセルRNA解析技術の出現により、膵癌の不均一性の解明が徐々に可能となってきた。本研究ではこれを用いて、膵癌と癌間質を構成する主要な細胞である癌関連線維芽細胞(CAF)の相互作用により化学療法に耐性を生じる機序を明らかにする。
【方法】
膵癌細胞株SUIT-2単培養もしくはSUIT-2+CAF共培養に対してGEM投与群、非投与群を作成（計4群)。これを72時間培養し、単一細胞懸濁液Chromiumでバーコーディングし、シングルセルRNA解析を行った。
【結果】
SUIT-2単培養非投与群で9272細胞、SUIT-2単培養GEM投与群で6890細胞、SUIT-2+CAF共培養非投与群で6665細胞、SUIT-2+CAF共培養GEM投与群で6237細胞が得られた。これらをmergeし、検出された全細胞に対しUMAPで次元圧縮を行い、CAFのクラスターを同定した。CAFを除去し癌細胞のみで再度UMAPを施行した結果、20425細胞において、10のクラスターに分けることができた。これらのクラスター間ではGEM投与により細胞分裂期の違いを認めた。また、GEMを投与することで出現したクラスターでは細胞障害の関連遺伝子が上昇する一方、代謝や免疫反応に関するクラスターや、分裂期のクラスターはGEM投与により減少していた。
【結語】シングルセル解析を用いて、癌細胞のGEM投与による変化について検討をおこなった。今後、CAFとの細胞間相互作用にも注目し、より詳細に治療抵抗性のメカニズムの解析を行う予定である。

Event date： 2022.7

Venue：横浜市   Country：Japan  

Event date： 2022.7

Venue：横浜市   Country：Japan  

潰瘍性大腸炎(UC)患者には慢性炎症の影響により、発病後7-8年で大腸がんが高頻度に生じる。遺伝的要因や腸内細菌叢の異常、自己免疫異常など様々な因子の関連が報告されているが、その病態はまだ明らかになっていない。腫瘍微小環境のうち、cancer associated fibroblasts(CAFs)は過去にmyofibroblast(myCAF)、inflammatory CAF(iCAF)、antigen-presenting CAF(apCAF)など、機能の異なる亜集団が存在しており、がんの浸潤や転移などの生物像に影響を及ぼすと報告されている。近年、大腸癌を含む様々ながん種でsingle cell RNA sequence (scRNA-seq)によるがん組織のheterogeneity解明が盛んに行われるようになったが、Colitis associated cancer(CAC)における腫瘍微小環境のheterogeneityはまだ解明されていない。scRNA-seqにより、同一UC患者内における大腸がん発生の様々な段階を単一細胞ごとに観察できると考えられる。今回我々は、当科にてUC由来大腸癌に対し大腸全摘術を行った1人の患者の正常部、炎症部、高度異形成部、癌部ｋオ・箇所より検体採取し、scRNA-seqを施行した。14936個の細胞の遺伝子発現データが得られ、クラスタリングにより19個の細胞集団が同定された。既知のマーカー遺伝子を用いてcell typeを同定した。腫瘍微小環境のうち特にfibroblastに着目し、fibroblast408細胞を抽出した。過去に報告のあるmyCAF、iCAF、apCAF関連遺伝子をスコア化し、検体採取部位別に比較した。また、それらをpublic dataを用いて通常型大腸癌と比較した。myCAFスコアは正常部・炎症部・高度異形成部には差がなかったが、癌部で上昇していた。iCAFスコアは癌部だけでなく高度異形成部でも低下していた。apCAFスコアは高度異形成部や癌部で低下しており、炎症部でも低下傾向が認められた。正常部と癌部の比較では、UC患者の各CAFスコアは通常型大腸癌と同じ傾向を示した。今回の検討で示された結果に基づき、UC由来癌の各発がん段階におけるfibroblastの変化について考察を加えて報告する。

Event date： 2022.7

Venue：横浜市   Country：Japan  

本邦における食道癌のうち85%以上が食道扁平上皮癌(ESCC)である。Stage II, IIIに対しては術前化学療法(NAC)後の外科的切除が標準治療とされているが、その5年生存率はそれぞれ56%, 29%と満足のいくものではない。一方近年、免疫チェックポイント阻害薬(ICB)の開発、適応が目覚ましく、ESCCに対してもICBの有効性が示されているが、その奏効率は20%ほどにとどまる。ICBの普及に伴い免疫学的側面から腫瘍微小環境を解明する試みも全世界的に加速しており、その手段として単一細胞レベルのmRNAを網羅的に解析するsingle cell RNA-sequence(scRNA-seq)の技術が盛んに用いられるようになってきている。本研究では、腫瘍微小環境における免疫環境の一端を担っている腫瘍浸潤マクロファージ(TAM)をscRNA-seqを用いて解析し、NACの有無で比較検討することで、NACがTAMにもたらす影響を解明することを目的とした。
当科においてESCCに対し手術を行った症例の摘出標本より正常食道粘膜および腫瘍部を採取しscRNA-seqを用いて細胞構成を同定し、その中で同定されたマクロファージ(Mp)集団を細分化して、それぞれの機能に着目して解析を行った。
正常部12症例、腫瘍部18症例の統合解析を?いCD45陽性細胞を抽出したところ、quality check後の細胞数は83,151で、11のクラスターに分割された。既知のcell typeに特徴的な遺伝子の発現パターンからそれぞれのクラスターが対応する既知のcell typeを同定し、その中からMpを抽出した。Mp (n=3,582)は4つに再クラスタリングされたが、既知のM1、M2マーカーでそのクラスターを特徴づけることは困難であり、差次的発現遺伝子からそれぞれが食作用、遊走、そしてサイトカイン分泌に特化したクラスターと、成熟したクラスターであることが示された。正常粘膜と腫瘍部を比較すると、腫瘍部では食作用の高い集団が著明に少なく、成熟した集団が極めて多い割合を占めていた。腫瘍部における成熟したTAMをNACの有無で比較したところ、活性化の程度には差がなかったが、走化性は有意に高く(p<0.001)、抑制性遺伝子の発現はNAC施行群で有意に低かった(p<0.001)。
以上から、NACがESCCのTAMの機能的なheterogeneityに変化をもたらしている可能性が示唆された。

Event date： 2022.7

Venue：横浜市   Country：Japan  

Event date： 2022.7 - 2023.7

Venue：函館市   Country：Japan  

Event date： 2022.7

Venue：Kyoto   Country：Japan  

Background
The goal of pancreatic cancer surgery is to obtain tumor-free margins (R0) and to perform a sufficient regional lymphadenectomy. To achieve these goals, radical antegrade modular pancreatosplenectomy (RAMPS) for pancreatic cancer has been widely accepted for cases with cancer at body and tail of the pancreas. However, some reports identified that medial approach of RAMPS in minimally invasive procedure is more difficult than that in open procedure due to the limitation of the operation view. We have developed an Gerota’s fascia identified “superior approach” to secure and identify the retroperitoneal borderline of the pancreas.
Method
Between April 2016 to December 2021, MIDP for PDAC was performed for 55 cases and the approach was performed from September 2019. We identify Gerota’s fascia as a landmark, because splenic artery (SPA), lymph node to be dissected, pancreas and tumor are all laid on the “cradle” of the Gerota’s fascia if there is no invasion of the tumor to the kidney. Therefore, if dissection was performed under Gerota’s fascia, the tissues to be dissected are all removed clearly. Approach from superior side is a useful way for detecting Gerota’s fascia and it is a “good entry” to the posterior plane of the fascia. Therefore, we start the dissection from superior side of the pancreas and widely pull the Gerota’s fascia off and remove all tissue to be dissected such as SPA, pancreas and lymph nodes and tumor
Results
Between 2019 November to 2021 September, Superior approach was performed in 16 cases (Robotic 4 cases and laparoscopic 12 cases). The median operation time was 276 min (154min to 467 min) and the median blood loss was 42.5g (0g to 619 g). All the cases were performed with pathological R0 resection.
Conclusion
The approach has been applied safely and will be a reasonable procedure for pancreatic cancer

Event date： 2022.7

Venue：Kyoto   Country：Japan  

Pancreatic ductal adenocarcinoma (PDAC) is a deadly disease, partly due to its resistance to chemotherapy. The mechanism of chemoresistance in PDAC involves the interaction between the high level of heterogeneity and abundant stroma cells, especially cancer-associated fibroblasts (CAFs), which is characteristic of PDAC and needs to be elucidated. To clarify it, we performed single-cell RNA sequence (scRNA-seq) analysis of PDAC cell-line that were cultured mono or co-culture with CAFs with Gemcitabine treatment.
As a result, there were 5496 viable cells of mono-culture group and 5251 cells of co-culture group. To analyze the PDAC cells only, we merged these cells into one data set and excluded CAFs. Using UMAP, a total of 10420 viable cells were identified with 7 distinct clusters. The proportion of specific three clusters was higher in co-culture group than that in mono-culture group. The ratio of G2M/S phase was higher in these clusters than that in other clusters, suggesting that cell division and DNA replication were active. In addition, these three clusters had unique pathways respectively: (1) pathways related to epithelial-mesenchymal transition, angiogenesis, and activin receptors, (2) pathways related to Rho GTP, and (3) HNF3B pathway and pathways that respond to drugs.
Here, we performed scRNA-seq analyses and investigated the effect of CAFs on PDAC cells under gemcitabine treatment, and present data suggest that CAFs may contribute to the more malignant behavior of PDAC cells even under chemotherapy, although further examination will be needed to elucidate the detailed mechanism of CAF-related chemo-resistance.

Event date： 2022.7

Venue：Kyoto   Country：Japan  

Pancreatic stellate cells (PSCs) are the primary stromal cells that interact with pancreatic tumour cells. PSCs transform from a quiescent state to activated myofibroblast-like cells with various stimuli including tumour-stromal interactions, such as direct contact with pancreatic cancer cells (PCCs) and growth factors secreted from PCCs. In turn, activated PSCs have reduced levels of lipid droplets containing vitamin A, increased α-smooth muscle actin (αSMA) expression, and secrete cytokines that promote proliferation and invasiveness of PCCs. Activated PSCs also produce extracellular matrix (ECM), which contributes to chemoresistance in cancer cells. Also, it has been reported that interactions with PSCs increase the malignancy of pancreatic cancer; therefore, attenuating the activation of PSCs may be a promising therapy for pancreatic cancer. However, little is known about the mechanisms of PSCs activation.
We have been investigated the mechanisms of PSC activation and also developed a new therapy for pancreatic cancer.
We have found, autophagy is related with activation of PSCs, and autophagy activation in PSCs is related with poor survival of patients with pancreatic cancer. Autophagy inhibition led PSCs to the quiescent state, which reduced the production of extracellular matrix and interleukin-6 in PSCs, followed by attenuated aggressiveness of pancreatic cancer in vitro. Autophagy inhibition in PSCs also reduced tumor enlargement, liver metastasis, and peritoneal dissemination in vivo. We found repressing autophagy in PSCs can be a promising therapy to inhibit the invasiveness of pancreatic cancer by decreasing tumor stromal interaction with PSCs.
We also have recently established a new high-throughput screening platform focusing on the morphologic feature of quiescent PSC. And the platform was able to visualize and quantify the activation of PSCs. This screening system is useful for discovering the compounds to attenuate PSCs activation and inhibit the progression of pancreatic cancer. We performed drug screening of the libraries of 3398 FDA-approved drugs with this platform and found that the accuracy of the screening system and also found that several compound could be promising candidate for new therapy for pancreatic cancer. Based on these findings, we hope the prognosis of pancreatic cancer would be improved in near future.

Event date： 2022.6

Venue：佐賀市   Country：Japan  

Event date： 2022.6

Venue：佐賀市   Country：Japan  

Event date： 2022.6

Venue：佐賀市   Country：Japan  

症例は55歳、女性。胃前庭部後壁の粘膜下腫瘍に対する術前の生検で胃NET,G3の診断であり、ロボット支援下幽門側胃切除及びD2リンパ節郭清術を施行したが、術後の病理診断では胃NEC、pT3(SS)pN3aM0;pStage3Bであった。WHO2010で消化管の神経内分泌腫瘍は、組織像によらずに増殖能(Ki-67指数)により、低値の場合は高分化腫瘍NET,G1(＜3%)、G2(3-20%)と称され、高値(＞20%)の場合は低分化癌NEC,G3に大別された。その後、形態学的に神経内分泌パターンを呈する高分化腫瘍NETでもKi-67が高値(>20%)の症例も多く報告され、低分化でKi-67指数が>20%のNECとは、予後、治療、バイオマーカーも異なることが判明し、WHO2019では、組織学的に神経内分泌パターンを示す腫瘍を高分化と称し、Ki-67指数が<3%、3-20%、>20%の判定により各々NET,G1、G2、G3と分類し、形態学的に低分化な神経内分泌腫瘍でKi-67指数が20%を超える腫瘍を・オ梵オ估睚・NECと分類することになった。本症例では術前の生検検体でKi-67指数が70%で比較的高分化な腫瘍部分が目立ち胃NET,G3と診断したが、外科切除検体では小細胞癌やLCNECと判断可能な組織像を認め、Rb及びp53の発現が消失していることから胃NECの最終病理診断に至った。胃NECは根治的切除が可能な局所領域病変であっても、手術単独での治療成績は極めて不良であり、手術単独療法は推奨されず、薬物療法や放射線療法を含めた集学的治療の一環として行うことが推奨されている。胃NECの抗腫瘍薬の選択には肺小細胞癌の治療に準じプラチナ系薬剤を含む併用療法が推奨されており、CDDP＋ETPやCDDP＋CPT-11を用いることが多い。今回、術前の生検検体で胃NET,G3であったが、外科切除検体の病理診断で胃NECと診断され、術後補助化学療法としてCDDP＋ETPの投与を行った1例を経験した。NET,G3およびNECに対する診断と治療方針に関して、若干の文献的考察を含めて報告する。

Event date： 2022.6

Venue：鳴門市   Country：Japan  

【背景】胃癌において、非充実型低分化型腺癌 (por2)は他のサブタイプより予後不良であり、一般的に豊富な繊維化組織を伴うことが多い。近年、膵癌や大腸癌でcancer-associated fibroblast (CAF)のheterogeneityがシングルセルRNAシーケシング (scRNA-seq)によって解明されているが、por2胃癌微小環境におけるCAFをscRNA-seqを用いて解明した報告はない。
【対象・方法】当科で胃切除術を施行した胃癌患者13例から採取した腫瘍部12例と正常部10例を対象とし、scRNA-seqを用いてpor2胃癌CAFが形成する免疫抑制環境について評価した。
【結論】全22症例の統合解析を行った結果、quality check後のCAF細胞数は2383であり、遺伝子発現の特徴からinflammatory CAF (iCAF)とmyoblastic CAF (myCAF)の2群に分類した。iCAFはmyCAFと比較して、免疫抑制機能関連遺伝子の発現が高値であり、Gene Set Enrichment Analysisではケモカイン産生や血管新生の遺伝子セット発現が有意に高値であった。por2胃癌のiCAFは、non-por2胃癌iCAFと比較して腫瘍促進やEMTに関連する遺伝子発現が有意に高値であった。一方、por2胃癌におけるTregの免疫抑制関連遺伝子の発現はnon-por2胃癌Tregと比較して有意に低かった。また、por2胃癌では細胞障害性T細胞の細胞障害性機能関連遺伝子の発現がnon-por2胃癌と比較して有意に低く、細胞障害性T細胞の割合もpor2胃癌で低かった (28.6 % vs. 35.6 %)。
【結論】por2胃癌の免疫抑制環境の形成にiCAFが重要な役割を果たしている可能性が示唆された。

Event date： 2022.6

Venue：千葉市   Country：Japan  

【Introduction】
The tumor microenvironment (TME) comprises a heterogeneous population of cancer-associated fibroblasts (CAF) with different functions: myofibroblastic CAF (myCAF), inflammatory CAF (iCAF), and antigen-presenting CAF (apCAF) in pancreatic and breast cancers. However, no study has investigated whether these CAF exist in gastric cancer (GC).
【Method】
We performed single-cell RNA sequencing analysis on 125898 cells from 12 tumors and 10 non-tumor samples from patients who underwent gastrectomy.
【Result】
We isolated the fibroblast cluster (n=2383) and identified four fibroblast clusters. The CAF marker genes in each cluster were cross-referenced with known markers of cell clusters to identify the different clusters such as my CAF, iCAF, and apCAF. We determined F0 and F2 clusters as iCAF, while F1 and F3 clusters as myCAF. The apCAF did not form a cluster but existed within the iCAF and myCAF clusters. To investigate the immune function of iCAF (F0 and F2) in GC, we used gene set enrichment analysis and found that four gene sets related to immunity, angiogenesis, and cytokine production are more enriched in iCAF cluster than in myCAF cluster. Furthermore, expression levels of immunosuppressive genes were highest in the F0 CAF cluster among all CAF clusters.
【Conclusion】
In the present study, we demonstrated that GC TME was composed of a heterogeneous population of CAF with different functions, as well as pancreatic and breast cancers.

Event date： 2022.5

Venue：San Diego  

【Introduction】
Myeloid-derived suppressor cells (MDSCs) suppress the immune response of cytotoxic T cells by inducing T cells to exhaust or fail to function. According to a previous report, there are two differentiation pathways from common myeloid progenitor to neutrophils and polymorphonuclear-MDSC (PMN-MDSC). However, a better understanding of PMN-MDSC differentiation remains to be established.
【Method】
We performed single-cell RNA sequencing analysis on 125898 cells from 12 tumors and 10 non-tumor samples from patients who underwent gastrectomy. We excluded the other cell types using canonical gene expression and doublet finder and isolated neutrophils and PMN-MDSC.
【Result】
We identified two neutrophil and three PMN-MDSC clusters, visualized with the UMAP algorithm. The analyses of neutrophil clusters revealed that P0, P1, and P4 expressed high immunosuppressive gene expression levels. These genes related to immunosuppressive function have been reported to be expressed in MDSC. Based on these findings, we defined P0, P1, and P4 clusters as PMN-MDSC. The percentage of PMN-MDSC was higher in the GC samples compared to the non-malignant samples (58.6 % vs. 46.9 %). We next used the Monocle analysis toolkit to perform cell trajectory analysis to investigate the potential transition between neutrophils and PMN-MDSC. The pseudotime trajectory axis indicated that neutrophils could transdifferentiate into PMN-MDSC. Furthermore, the cell cycle of PMN-MDSC had a lower percentage of G2M phase than that of neutrophils.
【Conclusion】
The present results suggest that local neutrophils can give rise to PMN-MDSC in gastric cancer.

Event date： 2022.5

Venue：San Diego  

【Introduction】
Myeloid-derived suppressor cells (MDSCs) suppress the immune response of cytotoxic T cells by inducing T cells to exhaust or fail to function. According to a previous report, there are two differentiation pathways from common myeloid progenitor to neutrophils and polymorphonuclear-MDSC (PMN-MDSC). However, a better understanding of PMN-MDSC differentiation remains to be established.
【Method】
We performed single-cell RNA sequencing analysis on 125898 cells from 12 tumors and 10 non-tumor samples from patients who underwent gastrectomy. We excluded the other cell types using canonical gene expression and doublet finder and isolated neutrophils and PMN-MDSC.
【Result】
We identified two neutrophil and three PMN-MDSC clusters, visualized with the UMAP algorithm. The analyses of neutrophil clusters revealed that P0, P1, and P4 expressed high immunosuppressive gene expression levels. These genes related to immunosuppressive function have been reported to be expressed in MDSC. Based on these findings, we defined P0, P1, and P4 clusters as PMN-MDSC. The percentage of PMN-MDSC was higher in the GC samples compared to the non-malignant samples (58.6 % vs. 46.9 %). We next used the Monocle analysis toolkit to perform cell trajectory analysis to investigate the potential transition between neutrophils and PMN-MDSC. The pseudotime trajectory axis indicated that neutrophils could transdifferentiate into PMN-MDSC. Furthermore, the cell cycle of PMN-MDSC had a lower percentage of G2M phase than that of neutrophils.
【Conclusion】
The present results suggest that local neutrophils can give rise to PMN-MDSC in gastric cancer.

Event date： 2022.5

Venue：佐賀市   Country：Japan  

ロボット支援下胃切除術は、郭清手技の自由度の高さやスムーズさに比べると、消化管再建手技はやや難しく感じる。その理由としては、ポート配置が腹腔鏡手術とは異なる点と、術者と助手の協働作業が必要になる点にあると考えられる。当科ではロボット支援下胃切除術におけるデルタ吻合を、腹腔鏡手術での手技と同じ手技で行えるように工夫している。
具体的には、術者が患者右側２本のアームを両手で操作できるように紐付けを変更し、助手が患者左側のアシストポートから自動縫合器を挿入し、胃壁を術者3rd arm（患者左側）で把持する。まず胃の小孔に助手が自動縫合機を挿入し、3rd armとともに十二指腸側にゆっくり移動した後は、そのまま静止する。そして、術者が２本のアームを用いて、自動縫合器に十二指腸をかぶせるように（靴下を履かせるように）しながら小孔に挿入する。
この手技は腹腔鏡手術と同じ感覚で行うことができ、かつロボット鉗子の自由度の高さや緻密さを生かせる点で有用だと考えられる。

Event date： 2022.5

Venue：佐賀市   Country：Japan  

Event date： 2022.5

Venue：佐賀市   Country：Japan  

家族性大腸腺腫症(FAP)はAPC遺伝子の生殖細胞系列遺伝子変異の影響で大腸癌が高頻度に生じる疾患で、標準術式は大腸全摘である。大腸全摘は剥離範囲が非常に広く、腸間膜の処理、再建など長時間手術が必要になる。低侵襲性からFAPに対する大腸全摘を腹腔鏡手術で行う施設が増えているが、さらなる長時間手術が問題になっている。経肛門的内視鏡手術は直腸癌手術で確立しつつある方法であり、経肛門もしくは会陰的に逆行性に直腸固有筋膜レベルでの剥離を行う手技である。腹腔操作と同時に行うことによって手術時間の短縮が可能になるだけでなく、2方向から確認を行いながらの剥離が可能であり、正確で安全な手術が可能になる。当科で施行したFAPに対する経肛門的内視鏡手術併用大腸全摘術は3症例で男性1例、女性2例で年齢は23.8 (18-29)歳であった。施行術式は全例大腸全摘であり、3例ともにHand-sewn IAAで、全例に回腸人工肛門を造設した。手術時間は590 (423-641)分であり、出血量は100 (0-200)gであった。Outlet obstructionが結オ梛ョ・隼廚錣譴Clavien-Dindo Grade 2の腸閉塞を1例に認めた。手術時間についてはやや長時間となっているが500分を超えた2例はともに腹部手術歴があり腹腔内癒着の剥離に時間を要したが安全に手術可能であった。本セッションでは2020年から当科で施行している経肛門アプローチ併用大腸全摘術の手術術式を供覧いただく。

Event date： 2022.5

Venue：佐賀市   Country：Japan  

Event date： 2022.5

Venue：佐賀市   Country：Japan  

症例は７８歳女性。胸部中部進行食道癌に対して鏡視下食道亜全摘、後縦隔胃管再建を施行した。術後経過は順調で退院後は外来で定期的な画像検査を行っていた。術後7年経過した際に、嘔吐と食事摂取不良、体重減少が出現し、精査を行った。内視鏡検査で胃内に悪性腫瘍をふくむ局所病変は認めなかったが、残渣が多量に貯留していた。造影CTでは胸腔内の胃管は多量の内容物で拡張しており横隔膜脚付近で管腔の狭小化を認め通過障害と考えられた。通過障害部位には硬い狭窄は認めず、食道裂孔レベルでの周囲組織からの圧迫に加え、加齢や胃壁の過度な膨張に伴う胃管の蠕動能の低下が原因と考えられた。絶食管理と経鼻胃管による減圧、蠕動促進薬の投与による保存的加療を行ったが、胃管の拡張は改善したものの通過障害の改善には至らなかった。そこで、通過障害部位に内視鏡的にカバードステントを留置することとし、ステント逸脱予防のためクリップで粘膜面より固定した。留置後しばらくは胃管拡張せず食事も再開できていたが、留置後1週間弱でクリップが外れステントは胸腔内へ逸脱した。ステントの固定がキ)タヌ・廚塙佑─▲好謄鵐販叡崕僂吠四僂靴栃・亢晴室蟒僂妊好謄鵐噺把蠅垢詈・砲箸靴拭・泙此・盪覿静・剖杭・・未縫・弌璽疋好謄鵐箸鯲叡屬靴人眛釗・盪覿世妊好謄鵐汎盥个魍稜Г靴疹・屬如∧・亢請犧遒砲曇勀賁未・薜瀛描漢悗鬚・韻織好謄鵐噺把蠅2針行い、同時に予防的に腸瘻造設を行った。術後、通過障害は認めず、栄養障害は改善し2週間程度で退院となった。現在術後2年経過しているが、ステントの逸脱は認めず有害事象なく経過している。近年では高齢化に伴い、術後通過障害の一因として加齢に伴う蠕動力低下が予想される。今回、ステント留置術に加え、より確実なステント固定として、腹腔鏡下にステントを全層固定したことが非常に有用であったと考えられた。若干の文献的考察をふまえて報告する。

Event date： 2022.5

Venue：佐賀市   Country：Japan  

Event date： 2022.5

Venue：佐賀市   Country：Japan  

症例は71歳、男性。9年前に喉頭癌に対して化学放射線療法を施行されたが、その2年後に再発を来したために、喉頭全摘・永久気管孔造設術を施行された。今回、吻合部を含む頚部～胸部食道に認められる多発食道癌に対して、当科紹介となり手術の方針とした。本症例は喉頭全摘術後であり、咽頭付近まで食道切除が必要であったが、可及的に長い胃管を作成することで咽頭胃管吻合が可能となり遊離空腸は不要であった。　胃管の作成時、右側の大網処理に際し、大網組織の血管系を損傷しないように慎重に処理して横行結腸をtake downし、胃及び十二指腸の受動を十分に行った。胃管の血行再建には短胃静脈を使用するため、腹部操作時には可能な限り長く損傷なく残すことに留意し、ヘモロックとハーモニックを使用して長い短胃静脈を3本温存した。胃管の作成は右胃動脈の分枝を確認し、2本残す部位で小弯の切離ラインを決定した。近年のICGによる血流評価の独自のデータの蓄積から、細径胃管としても血流に問題ないことが明らかになっており、本症例では幅３ｃｍの細径となるようにlin
ear staplerを用いて胃管を延長した。また、最初のstaplerは小弯から垂直に切り込むようにcurved cutterを使用し、これも胃管延長に有用であった。ICGを静脈投与し、胃管の先端まで血流がしっかりと存在していることを確認した。後縦隔経路で胃管を頚部まで挙上し、咽頭までの距離は十分であったため、咽頭胃管吻合を手縫いで行った。永久気管孔造設後の症例であったため後縦隔経路でも椎体と気管による胃管の圧排所見はなかったが、念の為、形成外科によるsuper chargeを頚横動脈-短胃動脈、super drainageを内頚静脈-短胃静脈で行った。当科における胃管再建の工夫について考察し、報告する。

Event date： 2022.5

Venue：佐賀市   Country：Japan  

Event date： 2022.4 - 2022.5

Venue：Tampa  

Event date： 2022.4

Venue：熊本市   Country：Japan  

本邦における食道癌のうち85%以上が食道扁平上皮癌(ESCC)である．その中でもStage II, IIIについては術前化学療法(NAC)後の切除が標準治療である．近年，固形癌に対する免疫チェックポイント阻害薬(ICB)の効果が次々と実証され，ESCCについても二次治療としてICBが適応となったが，その奏効率は20%程度に留まる．腫瘍が発生し制御できなくなってしまうことや腫瘍が治療抵抗性を持ってしまうことには多くの原因が存在しうるが，腫瘍微小環境(TME)における免疫状態の変化もその一因として極めて重要と考えられる．そこで今回，術前化学療法(NAC)が，ESCCのTMEの中でも樹状細胞(DC)にもたらす変化を解明することを目的として解析を行った．
当科にてESCCに対し手術加療を行った症例の摘出標本よりその腫瘍部を採取し，single-cell RNA-sequenceを用いて腫瘍組織の細胞構成，およびその同定された細胞集団ごとの機能に関して解析し，NACの有無で比較検討を行った．DCは，NAC施行9症例より718細胞，非NAC施行9症例で394細胞得られ，5つのクラスターに大別された．それぞれのクラスターの機能を解析したところ，NAC施行群では，cDC1クラスターは活性化シグネチャが有意に高いこと(p<0.01)，cDC2クラスター，moDCクラスターは有意に機能不全シグネチャが低いこと(p<0.01)がそれぞれ示された．
以上より、NACがESCCのTMEにおけるDCに機能的な変化をもたらしている可能性が示唆された。

Event date： 2022.4

Venue：熊本市   Country：Japan  

【背景】膵癌(PDAC)は予後不良な腫瘍であり、5年生存率が8％未満で致命的な癌の1つである。Acinar-to-ductal metaplasia (ADM)は、膵癌の前癌病変の一時的な状態であり、さらに低悪性度の前癌病変になり、膵癌に進展すると言われている。我々は以前に、cancer-associated ADM(CA-ADM)が線維形成性変化と局所膵実質への癌細胞の浸潤を促進することを明らかにした、さらに、IL12Aが正常な腺房細胞と比較してCA-ADMでより高く発現することも報告している(Kibe et al、Cancer Letter、2019)。一方で、IL12Aは、膵癌の血管新生につながる単球系統細胞を動員することが報告されており、血管新生は腫瘍進展における重要なイベントであることが知られている。ただし、IL12AがCA-ADMにおける血管新生機序に関わっているかは不明である。
【目的】PDAC進展過程におけるCA-ADMや同部の血管新生の意義について、その癌微小環境に着目し検討する。
【方法】ヒト膵癌切除組織131例を用いて、CD31、Amylase、CK19、CD68、CD163、IL12Aの免疫組織染色を行なった。KPC (KrasLSL-G12D/+;Trp53LSL-R172H/+;Cre)マウス膵癌切除36例をCD34、AmylaseとCK19の免疫組織染色を行なった。さらに、ヒト膵癌切除組織5例の多重免疫組織染色を行い、CD68、CD163とMMP9の発現と分布を検討し、CA-ADMにおける微小血管密度とtumor-associated macrophage(TAM)の関連を評価した。また、単球細胞株THP-1をマクロファージ細胞に分化誘導し、Western BlotとRT-PCRでMMP9の発現量を測定した。
【結果】ヒトCA-ADMの微小血管密度はPDACに比べ顕著に高かった。また、CA-ADMについて微小血管密度が高いほど予後不良であった。さらに、CD68とCD163、IL12Aを染色し、M2型を含むTAMとIL12Aの発現を検討した結果、CA-ADMのCD68、CD163とIL12AはPDACに比べ顕著に高かった。多重染色によって、TAMのM2型はMMP9を分泌することが明らかになった。一方、KPCマウスCA-ADMの微小血管密度はPDACに比べて高かった。ただし、微小血管密度と生存率が関係なかった。
【考察】ヒトCA-ADMの微小血管密度は豊富である。TAMはCA-ADMの血管新生に関与している可能性がある。一方、マウスではヒトの結果とは一致しておらず、その原因を明らかにするためには検討が必要である

Event date： 2022.4

Venue：熊本市   Country：Japan  

【はじめに】横行結腸癌に対する腹腔鏡下手術は,郭清手技を含めて難易度が高いとされている.郭清範囲やアプローチ方法も腫瘍局在や施設間で異なり,特に横行結腸中央部癌に対して,結腸半側切除(HC)か結腸部分切除(PC)かの選択について明確な基準がない.今回当科で施行した腹腔鏡下手術について,腫瘍局在や術式選択による術後成績への影響を検証した.
【方法】2010年1月から2020年12月まで,当科で腹腔鏡下手術を施行した84例(Stage?-?,他臓器合併/同時切除症例除く)を対象とした.患者背景,腫瘍局在と術式,術後合併症率等を後方視的に検討した.局在は右側(R)/中央(M)/左側(L)群に,術式はHC群(右:RHC/左:LHC),PC群に分類した．
【結果】腫瘍局在はR群40例(48%)/M群17例(20%)/L群27例(32%)で，術式はR群でRHC29例/PC11例,M群でRHC4例/PC12例,L群でRHC1例/PC21例/LHC5例が行われた.HC群とPC群で年齢,性別,BMIに差はなかった.リンパ節転移はR群で13例(33%)に認め2例は回結腸(ICR)と中結腸動脈(MCA)領域に,残り11例はMCA領域のみに,M群では8例(47%)で全てMCA領域に,L群は9例(33%)で7例はMCA領域のみ,2例は左結腸動脈領域のみの転移であった.手術成績をHC(40例)とPC(44例)群で比較すると,手術時間(318vs298分),出血量(69vs54ml)に差はなかった．術後在院日数(12.3vs16.4日,p=0.11)はPC群でやや長い傾向にあった.Clavien-Dindo分類Grade3以上の合併症率には差を認めなかったが，Grade2以上はPC群(18例)で有意に多く(p&lt
;0.01),そのうち7例がイレウスであった.R群で5例,M群1例,L群4例に再発を認め,L群の1例のみSMA腹側の局所リンパ節再発であったが,高齢のためPC+D2郭清に止めた症例(pT3N2a)であった.
【まとめ】局在別の術式・リンパ節郭清範囲は許容できると考えられた.PC群はやや合併症が多く在院日数が長くなる傾向にあり,高齢者などハイリスク患者への適応は慎重に行う必要がある.今後は長期成績・予後についての検討必要である.

Event date： 2022.4

Venue：熊本市   Country：Japan  

日本における膵癌の約95％が腺がんである。その5年生存率は男性8.9%、女性8.1%、従来療法の治療効果は十分ではない。近年、免疫チェックポイントインヒビターは様々な癌種に対して治療効果を示しているが、膵癌に対しては効果的ではない。膵癌は免疫学的に「コールド」腫瘍微小環境(TME)を示し、CD8+ T細胞の低浸潤とGranzyme-BやIFN-γなどの低発現、骨髄由来細胞の高浸潤を特徴とする。
栄養競争は「コールド」TMEの原因の一つとして考えられている。グルコース代謝は免疫細胞において重要で、その生存と機能を制御している。KRAS変異（膵癌は95％以上）によって癌細胞は解糖系を亢進させ、TME中のグルコースを消耗する。同時に、微小環境中のPD-L1とPD-1の結合で癌細胞の解糖系を亢進し、結果として、TME中の乳酸濃度が上昇し、免疫抑制性のTMEの原因になる可能性が示唆されている。樹状細胞(DC)は抗原提示細胞として、T細胞の活性化に重要な役割を果たしているが、その成熟には急速な解糖系の亢進が必要でさらにその機能維持にもその解糖状態を保つ必要がある。T細胞はその活性化と抗腫瘍作用において糖代謝に依存している。
膵癌におけるコールド微小環境の形成に、微小環境中のグルコース代謝の異常がどのように影響するかは明らかになっていない。
【目的】腫瘍微小環境におけるグルコース代謝が抗腫瘍免疫に与える影響を検討する。
【方法】KPC(KrasLSL-G12D/+;Trp53LSL-R172H/+;Cre)マウス由来の癌細胞から各グルコース消耗率(高グルコース代謝、Control、低グルコース代謝)の細胞株を樹立し、B6マウス(C57BL/6)に同所と皮下に移植しすることで低グルコースTMEモデル、Control TMEモデル、高グルコースTMEモデルを樹立して、Flow cytometryとIHCにて免疫微小環境の変動を検討した。また、オルガノイドを用いた癌細胞とDCの 3D共培養実験でCD80、CD86、MHC I、MHC IIの発現及び細胞数を検討した。
【結果】低グルコースTMEの移植モデルはControlより低CD4＋T細胞、低CD8＋T細胞、低DCの浸潤が減少していた。3D共培養実験では低グルコースTMEのモデルはControlよりDCの細胞数が減少し、低グルコースTMEの中ではDCの生存が抑制されることが示唆された。また、CD86、MHC I、MHC II発現が低下していた。
【考察と結論】今後、さらなる検討が必要であるが、膵癌微小環境中のグルコース代謝を選択的に制御することで抗腫瘍免疫を増強できる可能性があると考えられる。

Event date： 2022.4

Venue：熊本市   Country：Japan  

＜はじめに＞
鏡視下膵頭十二指腸切除術（MIPD）においてSMAへのアプローチに関して複数の報告がある。昨年本邦で行われた”安全な肝胆膵MISのためのPrecision Anatomyに関するコンセンサスミーティング”でも右側からのアプローチは知っておくべきであるが、様々なアプローチを取り入れるべきであるというコンセンサスであった。当科では膵頭部左側から膵頭部背側を授動、SMAへのアプローチを行う独自のアプローチを用いており、ビデオを供覧しながらその手技を紹介する。
＜手術手技＞
・膵頭部背側授動；いわゆる患者右側からの＜kocher＞の授動は用いていない。横行結腸間膜を頭側に翻転し空腸起始部を確認する。空腸とIMV間の腹膜を剥離、空腸左側の剥離により下大静脈を同定、下大静脈前面を露出させる。その後頭側に向かって剥離、左腎静脈を確認、十二指腸右側まで膵頭部背側を剥離する。
・SMAアプローチ；空腸を扇型に展開、空腸間膜を切開し、1stJAを同定する。1stJA前面に沿って小腸間膜を中枢側に向かって剥離、共通幹から分岐するIPDAを同定、これをクリップし、膵頭部の血流遮断を行っておく。症例が膵癌の場合には1stJAも切離しておく。その後は右側からのアプローチで右側からのアプローチでIPDV、IPDAの処理を行う。膵を切離後、SMV右側を剥離、膵上縁でPSPDVを確認、標本を摘出する。
＜結果＞
2016年7月から2021年8月までの間にMIPD 58例（腹腔鏡下34例、ロボット支援下24例）施行した。手術時間中央値、出血量中央値はそれぞれ690分 (p10-p90,445-838分)、347ml(p10-p90,81-976ml)であった。術後在院死亡は認めてない。
＜結語＞
ロボット支援下PDの保険収載に伴い、腹腔鏡、ロボット支援下手術いずれでも利用できる手技での定型化をおこなった。今後さらに定型化を進め、安全なMIPDの普及に勤めたい。

Event date： 2022.4

Venue：熊本市   Country：Japan  

【背景】
癌の進展において、癌細胞とその周囲組織から構成される腫瘍微小環境が重要な役割を果たしているが、特に近年、免疫チェックポイント阻害薬の出現によって腫瘍内の多様な免疫細胞で構成される免疫微小環境の不均一性が注目されている。一方、cisplatinやgemcitabineなどの化学療法により免疫微小環境が大きく変動することが明らかになっている。今回、食道癌の手術切除サンプルを使用したscRNA-seqにより腫瘍浸潤B細胞の遺伝子発現解析を行うことで、術前化学療法(NAC)によるB細胞の不均一性の変動解析を行った。
【対象】
当科で外科的切除を行った食道扁平上皮癌において、NAC施行例 (4例)、NAC未施行例(5例)、正常食道粘膜部(7例)の計16検体を対象とした。検体は単一細胞化したのちにchromiumを使用しscRNA-seqを行い腫瘍浸潤B細胞の発現遺伝子の変化を評価した。
【結果】
計16検体のシークエンスデータに対しSeuratパッケージを使用しQuality Checkを施行すると全細胞数：89269、総遺伝子数：29331が検出された。UMAPで次元圧縮を行い有意発現遺伝子でクラスター化すると19のクラスターに分けられた。さらに各クラスターの発現遺伝子からＢ細胞を分離し、再クラスター化を行うと9のクラスターに分類でき、NAC施行群のB細胞総数は1981個、NAC未施行群は2200個であった。Ｂ細胞の各サブタイプの代表遺伝子の発現で評価すると、Naive B細胞、メモリーB細胞などのクラスターが同定できた。Naive B細胞に注目して、代表的な活性化受容体であるIgDを評価したところ、NAC施行群で著明に発現が低下していることがわかった。
【結語】Naive B細胞がNAC施行に伴って機能低下している可能性が示唆された。今後T細胞や樹状細胞などの多様な免疫細胞との関係を考慮しながら解析をすすめていく予定である。

Event date： 2022.4

Venue：熊本市   Country：Japan  

Event date： 2022.4

Venue：熊本市   Country：Japan  

末期腎不全に対する血液・腹膜透析の代替手段として腎移植は海外で広く行われており、国内でも生体腎移植が増加傾向にある。移植によって透析から離脱できるだけでなく、移植により長期予後の改善も期待できる。腎移植の増加による末期腎不全患者の予後改善により大腸癌に罹患する腎移植患者は今後も増えてくると考えられる。長期間使用する必要のある免疫抑制剤が大腸癌に与える影響も懸念される。今回我々は2008年～2021年に当院で大腸癌に対する外科手術を施行した腎移植後大腸癌6例について臨床病理学的因子を検討した。男女比は3：3で年齢は56.8 (42-70)歳、腎移植から大腸癌手術までの期間は95.5 (24-180)ヶ月、移植までの透析期間は17.2 (0-36)ヵ月であった。免疫抑制剤は
ミコフェノール酸モフェチルを5例、タクロリムス5例、メトロプレドニゾロン6例、シクロスポリン1例、アザチオプリン1例を使用していた。癌の局在は右側結腸3例、横行結腸1例、直腸2例であった。クローン病に合併する痔瘻癌と高度の腹膜播種を認めた2例は人工肛門造設のみであったが、残り4例は腹腔鏡で根治手術が可能であった。術後合併症は1例に縫合不全、脳梗塞を発症した。病理組織診断では分化型腺癌が4例、印環細胞を伴う低分化腺癌が1例、扁平上皮癌が1例であった。切除可能であった4例ではT2 1例、T3 3例であり、リンパ節転移陽性を1例に認めた。長期予後としては切除可能な症例では4例中1例で早期の肺転移再発を来していた。小数例の検討であり十分な検討ができているとは言い難いが移植後大腸癌では扁平上皮癌や印鑑細胞癌などの特殊型が多い可能性が示唆された。

Event date： 2022.4

Venue：熊本市   Country：Japan  

Event date： 2022.4

Venue：熊本市   Country：Japan  

【背景】進行胃癌の再発率を改善するため、従来の術後化学療法に加えて、術前化学療法(NAC)について議論されている。本邦においてはbulky Nに対するNACの有用性が報告されたが、まだ確立された治療戦略とは言えない。
【目的】当科における切除可能進行胃癌に対するNAC施行例を後ろ向きに検討し、特に再発例の臨床病理学的特徴について考察した。
【対象】2010年以降、進行胃癌・食道胃接合部癌に対してNACを1コース以上施行後に根治手術を行った24例について検討した。ただし、Conversion手術例は含まず、手術時にCY1などの非治癒因子を認めた症例(Stage IV)は除外した。
【結果】24例中11例(46%)に再発を認めた。治療前の病態により以下の3群に分けて、その特徴を検討した。(1) bulky Nまたは多発リンパ節転移(cN2-3)では14例中6例(43%)に再発を認めた(リンパ節再発2例、肝転移2例、後腹膜再発2例）。再発は全て2年以内に認められ、うち4例は術後6か月以内の再発であった。(2) 腫瘍径8cm以上の巨大3型進行癌では7例中2例（29%)に再発を認め、1例は術後3か月（肝転移）、もう1例は術後1年半（後腹膜）での再発であった。(3) 4型進行癌では4例中3例(75%)に再発を認め（腹膜転移2例、局所再発1例）、1例は他病死であった(術後2か月)。再発は術後1年-3年半までに認められた。ただし、これ以外にConversion手術後(CY1→CY0)に術後6年無再発生存例がある。なお、食道胃接合部癌では10例中5例に再発を認めた(術前bulky N 4例、巨大腫瘍1例)$
B!#
【考察】bulky N・多発リンパ節転移群では予後良好な症例がある一方で、再発までの期間が短く、術後早期の化学療法が必要と考えられた。4型進行癌はNACが無効であることが多く、現時点では早期の手術を考慮すべきであると考えられた。
【結論】進行胃癌に対しては、治療前の病態によって術後再発率や再発までの期間が異なることに留意した治療戦略が求められる。

Event date： 2022.4

Venue：熊本市   Country：Japan  

[背景] 膵癌は、膵癌は豊富な間質を特徴とし、その癌間質から誘導される微小環境因子が癌の増殖や転移に寄与するとされる一方で、癌間質を減少させると低分化型膵癌が増え予後が悪化したとの報告も認める。予後や治療反応性を左右するphenotypeを規定し得る膵癌間質の癌細胞への影響を明らかにすることは、膵癌における新たな治療戦略を構築する上で重要であると考えられるが、依然として不明な点は多い。
[目的] 微小環境因子依存性に基づいた膵癌サブタイプ分類を行い、発現プロファイルに応じた薬剤反応性の差違を明らかにする。
[方法] ヒト膵癌組織より膵癌オルガノイドを樹立し、網羅的遺伝子発現解析を行った。各膵癌オルガノイドの微小環境因子への依存性を評価し、微小環境因子依存性に基づいて<high>、<low>の2群に分けた。微小環境因子依存性と相関する遺伝子発現を評価し、各群において有効と考えられる薬剤を選出した。これら薬剤を用いたviability assayを行い、微小環境因子依存性と薬物反応性の相関を評価した。
[結果] 8例の膵癌オルガノイドを樹立した。これらオルガノイドは網羅的発現データを基に二つのクラスターに分類され、各クラスターは微小環境因子依存性に基づいた<high>、<low>の2群にそれぞれ対応していた。微小環境因子依存性と相関して変動する遺伝子をGene Ontology解析によって検索し、<high>群ではメバロン酸代謝関連遺伝子の発現が増加しており、一方で<low>群では細胞周期関連遺伝子の発現が増加していた。SimvastatinとGemcitabineを用いてviability assayを行ったところ、＜high＞群ではSimvastatinに対する感受性が有意に高く、＜low＞群ではGemcitabineに対する感受性が有意に高かった。
[結語] 膵癌オルガノイドは、微小環境因子依存性によってそれぞれ異なる発現プロファイルと薬剤反応性を示した。これらの結果は、今後の個別化治療や新たな標的治療を開発する上で重要な知見となり得ると考えられた。

Event date： 2022.4

Venue：熊本市   Country：Japan  

Event date： 2022.4

Venue：熊本市   Country：Japan  

腫瘍微小環境において、CD8+Tcellは樹状細胞などの抗原提示細胞からの刺激を受けて活性化し、がん細胞を直接攻撃、殺傷するなどして、腫瘍免疫の中心を担っている。家族性大腸腺腫症(FAP)患者にはAPC遺伝子の生殖細胞系列遺伝子変異の影響で大腸癌が高頻度に生じ、様々な段階の前癌病変も存在する。近年、single cell RNA sequence (scRNA-seq)によるがん組織のheterogeneity解明が盛んに行われるようになったが、FAPにおける腫瘍微小環境のheterogeneityはまだ解明されていない。
今回我々は、当科にてFAPに対し手術を行った1人の患者から、正常部、腺腫非密生部、腺腫密生部、癌部の4検体を採取し、10X chromium systemを用いてscRNA-seqを施行した。33266個の細胞の遺伝子発現データが得られた。Seurat上で解析を行い、UMAPを作成したところ，22個のクラスターが得られた。既知のマーカー遺伝子を用いてcell typeを同定し、CD8+Tcellのみを抽出して再クラスタリングを行ったところ、8個のクラスターが得られた。Na$(D+Ave、Cytotoxicity、Exhaustionに関連した遺伝子発現から、サブクラスタリングを行い、検体採取部位間でCD8+Tcellの構成を比較した。腺腫密生部、癌部ではCytotoxic effector CD8+ Tcellの割合が高いという結果が得られた。また、Exhaustionに関連した遺伝子の発現は、癌部だけでなく腺腫部でも上昇していた。
今回の検討で示された結果に基づきFAPの各発癌段階におけるCD8+Tcellのはたらきについて考察を加えて報告する。

Event date： 2022.3

Venue：Houston  

【Introduction】
Gastric non-solid type poorly differentiated adenocarcinoma (por2) has a poorer prognosis than other types of gastric cancer (GC). No studies have evaluated the function of CD8-positive T cells in por2 GCs using single-cell RNA sequence (scRNA-seq).
【Method】
We performed scRNA-seq analysis on 125898 cells from 12 tumors (including eight por2 GC) and ten non-tumor samples from patients who underwent gastrectomy.
【Result】
CD8-positive T cell cluster (n=11615) was extracted and reclassified into six clusters with different functions, classified based on RNA of T-cell-related genes. This process allowed the definition of five major populations: terminally exhausted, progenitor exhausted, acutely activated, γδT like and memory CD8-positive T cells. Por2 GCs were significantly less cytotoxic and exhausted than non-por2 GCs (p <0.001). Additionally, the proportion of cytotoxic T cells (28.6 % vs. 35.6 %) and naive T cells (23.6 % vs. 28.8 %) was lower in por2 GCs than in other GCs. The trajectory inferred using monocle3 indicated that exhausted T cells may follow a different pathway of differentiation from memory T cells and acutely activated T cells.
【Conclusion】
In the present study, we revealed CD8-positive T cells heterogeneity in GCs. Furthermore, we suggest that por2 GCs may be associated with a poor prognosis due to the less cytotoxic function of CD8-positive T cells.

Event date： 2022.3

Venue：神戸市   Country：Japan  

Event date： 2022.3

Venue：横浜市   Country：Japan  

【Background】
Gastric poorly differentiated adenocarcinoma has a poorer prognosis than other types of gastric cancer (GC), and non-solid type poorly differentiated adenocarcinoma (por2) generally has abundant tumor stroma. Among these various components in the tumor stroma, cancer-associated fibroblasts (CAFs) have been suggested to play a key role in tumor development. However, the role of CAFs in the tumor immune microenvironment (TIME) has rarely been reported in GC. Furthermore, to the best of our knowledge, no study has evaluated the heterogeneity of CAFs in GC. To reveal the heterogeneity of CAFs and TIME in GC, we focused on CAFs in por2 GC using single-cell RNA sequence (scRNA-seq).
【Method】
We performed scRNA-seq analysis on 125898 cells from 12 tumors (including eight por2 GC) and ten non-tumor samples from patients who underwent gastrectomy.
【Result】
Fibroblast cluster (n=2383) was extracted and reclassified into five clusters with different functions. In the ACTA2-negative CAF cluster, por2 GCs were significantly more capable of cancer promotion (p=0.017) and epithelial-mesenchymal transition (p<0.001) compared with non-por2 GCs. Additionally, in cytotoxic CD8 T cells, por2 GCs were significantly less cytotoxic than non-por2 GCs (p <0.001). Furthermore, the ratio of regulatory T cell counts/ cytotoxic CD8 T cell counts was significantly higher in por2 GCs. (p =0.011).
【Conclusion】
In the present study, we revealed CAFs heterogeneity in GC. Furthermore, we suggest that por2 CAFs may play an essential role in the formation of tumor-promoting TIME.