Kyushu University Academic Staff Educational and Research Activities Database
List of Papers
Nokitaka Setsu Last modified date:2020.05.29

Assistant Professor / Orthopaedics, Kyushu University School of Medicine / Department of Clinical Medicine / Faculty of Medical Sciences

1. Yoshihiro Matsumoto, Kenichi Kawaguchi, Jun ichi Fukushi, Makoto Endo, Nokitaka Setsu, Keiichiro Iida, Satoshi Baba, Hirokazu Saiwai, Akinobu Matsushita, Mitsumasa Hayashida, Seiji Okada, Yasuharu Nakashima, Clinical outcome and prognostic factors of malignant spinal dumbbell tumors, Spine Surgery and Related Research, 10.22603/ssrr.2018-0004, 2, 4, 317-323, 2018.01, Introduction: To investigate the clinical outcome and prognostic factors of malignant spinal dumbbell tumors (m-SDTs). Methods: We retrospectively reviewed the clinical outcome of 22 consecutive cases of m-SDTs and analyzed the prognostic factors associated with worse outcome. Results: Nineteen of the 22 cases were managed with surgery (86%), and gross total resection (GTR) was achieved in four cases (21%). The duration of overall survival (OS) ranged from 3 to 140 months, with a median survival time of 15.3 months. The 5 year OS rate was 55.6%. In multivariate analysis, histological subtype (high-grade malignant peripheral nerve sheath tumor) (hazard ratio [HR] 14.9, p = 0.0191), GTR (HR 0.07, p = 0.0343), and presence of local recurrences (HR 11.2, p = 0.0479) were significant and independent predictors of OS. Conclusions: On the basis of clinical data, we propose that GTR and prevention of local recurrence may improve the clinical outcome of m-SDTs..
2. Makoto Nakagawa, Fumihiko Nakatani, Hironori Matsunaga, Takahiko Seki, Makoto Endo, Yoko Ogawara, Yukino Machida, Takuo Katsumoto, Kazutsune Yamagata, Ayuna Hattori, Shuhei Fujita, Yukiko Aikawa, Takamasa Ishikawa, Tomoyoshi Soga, Akira Kawai, Hirokazu Chuman, Nobuhiko Yokoyama, Suguru Fukushima, Kenichiro Yahiro, Atsushi Kimura, Eijiro Shimada, Takeshi Hirose, Toshifumi Fujiwara, Nokitaka Setsu, Yoshihiro Matsumoto, Yukihide Iwamoto, Yasuharu Nakashima, Issay Kitabayashi, Selective inhibition of mutant IDH1 by DS-1001b ameliorates aberrant histone modifications and impairs tumor activity in chondrosarcoma, Oncogene, 10.1038/s41388-019-0929-9, 38, 42, 6835-6849, 2019.10, Chondrosarcoma is the second most common malignant bone tumor. It is characterized by low vascularity and an abundant extracellular matrix, which confer these tumors resistance to chemotherapy and radiotherapy. There are currently no effective treatment options for relapsed or dedifferentiated chondrosarcoma, and new targeted therapies need to be identified. Isocitrate dehydrogenase (IDH) mutations, which are detected in ~50% of chondrosarcoma patients, contribute to malignant transformation by catalyzing the production of 2-hydroxyglutarate (2-HG), a competitive inhibitor of α-ketoglutarate-dependent dioxygenases. Mutant IDH inhibitors are therefore potential novel anticancer drugs in IDH mutant tumors. Here, we examined the efficacy of the inhibition of mutant IDH1 as an antitumor approach in chondrosarcoma cells in vitro and in vivo, and investigated the association between the IDH mutation and chondrosarcoma cells. DS-1001b, a novel, orally bioavailable, selective mutant IDH1 inhibitor, impaired the proliferation of chondrosarcoma cells with IDH1 mutations in vitro and in vivo, and decreased 2-HG levels. RNA-seq analysis showed that inhibition of mutant IDH1 promoted chondrocyte differentiation in the conventional chondrosarcoma L835 cell line and caused cell cycle arrest in the dedifferentiated JJ012 cell line. Mutant IDH1-mediated modulation of SOX9 and CDKN1C expression regulated chondrosarcoma tumor progression, and DS-1001b upregulated the expression of these genes via a common mechanism involving the demethylation of H3K9me3. DS-1001b treatment reversed the epigenetic changes caused by aberrant histone modifications. The present data strongly suggest that inhibition of mutant IDH1 is a promising therapeutic approach in chondrosarcoma, particularly for the treatment of relapsed or dedifferentiated chondrosarcoma..
3. Yoshihiro Matsumoto, Akira Matsunobu, Kenichi Kawaguchi, Mistumasa Hayashida, Keiichiro Iida, Hirokazu Saiwai, Seiji Okada, Makoto Endo, Nokitaka Setsu, Toshifumi Fujiwara, Shingo Baba, Satoshi Nomoto, Yasuharu Nakashima, Clinical results of carbon-ion radiotherapy with separation surgery for primary spine/paraspinal sarcomas, International Journal of Clinical Oncology, 10.1007/s10147-019-01505-y, 24, 11, 1490-1497, 2019.11, Purpose: To evaluate the clinical outcome of combination of carbon-ion radiotherapy with separation surgery (CIRT-SS) in patients with primary spinal/paraspinal sarcoma (PSPS) and epidural spinal cord compression (ESCC). Methods: CIRT-SS was performed in 11 consecutive patients. Patients treated in the primary and salvage settings were categorized into Group A (n = 8) and Group B (n = 3), respectively. Clinical results and imaging findings were collected, with a particular focus on ESCC grade, treatment-associated adverse events (AEs), and the locoregional control (LRC) rate and overall survival (OS). Results: The median follow-up period from the start of CIRT-SS was 25 months (7–57 months). ESCC was improved by SS in all cases. No patients exhibited radiation-induced myelopathy (RIM), but three developed Grade 3 vertebral compression fracture (VCF) during follow-up. Locoregional recurrences were observed in four patients [Group A: 1 (12.5%), Group B: 3 (100%)]. Over the entire follow-up period, three patients developed distant metastases and two patients died. The 2-year LRC rate and OS were 70% and 80%, respectively. Conclusion: CIRT-SS in the primary setting achieved acceptable LRC and OS without RIM in patients with PSPS and with ESCC. VCF was the most frequent AE associated with CIRT-SS..
4. Amanda R. Dancsok, Nokitaka Setsu, Dongxia Gao, Jean Yves Blay, David Thomas, Robert G. Maki, Torsten O. Nielsen, Elizabeth G. Demicco, Expression of lymphocyte immunoregulatory biomarkers in bone and soft-tissue sarcomas, Modern Pathology, 10.1038/s41379-019-0312-y, 32, 12, 1772-1785, 2019.12, Despite advances in our understanding of the underlying molecular drivers of sarcomas, few treatments are available with proven benefit for advanced metastatic sarcomas. Immunotherapy has value in this setting for some types of cancers, but sarcomas, with their multiplicity of rare types, have not been characterized in detail for their expression of targetable immune biomarkers. This study provides the most systematic evaluation to date of tumor-infiltrating lymphocytes and immune checkpoint biomarker expression in sarcomas. We examined by morphology and immunohistochemistry 1072 sarcoma specimens representing 22 types, in addition to 236 benign bone and soft-tissue tumors. Genomically-complex sarcoma types—those driven by mutations and/or copy-number alterations—had much higher numbers of tumor-infiltrating lymphocytes than translocation-associated sarcomas. Prior exposure to radiotherapy was associated with increased immune infiltrates. Higher lymphocytic infiltration was associated with better overall survival among the non-translocation-associated sarcomas. Expression of PD-1 and CD56 were associated with worse overall survival. LAG-3 and TIM-3, two emerging immune checkpoints, were frequently expressed in most sarcoma types. Indeed, most cases positive for PD-(L)1 coexpressed one or both of these novel biomarkers, providing a potential rationale in support for trials targeting LAG-3 and/or TIM-3 in conjunction with PD-1 inhibition..
5. Noriko Oyama, Kanako Kojima-Ishii, Naoko Toda, Terumichi Matsuo, Vlad Tocan, Kazuhiro Ohkubo, Utako Oba, Yuhki Koga, Nokitaka Setsu, Yuichi Yamada, Kenichi Kohashi, Yasuharu Nakashima, Yoshinao Oda, Kenji Ihara, Shouichi Ohga, Malignant transformation of phosphaturic mesenchymal tumor
A case report and literature review, Clinical Pediatric Endocrinology, 10.1297/cpe.29.69, 29, 2, 69-75, 2020.01, Phosphaturic mesenchymal tumor, mixed connective tissue variant (PMT-MCT) causes tumor-induced osteomalacia (TIO). Most cases follow a benign clinical course, with rare occurrences of malignant transformation. We report a case of malignant PMT-MCT and review previous malignant cases to identify predictive factors for transformation. A 13-yr-old female, who presented with hypophosphatemic rickets, elevated serum intact fibroblast growth factor 23 (FGF23) levels, and a nodule in the back, received a diagnosis of TIO because of the benign PMT histopathology. After resection of the primary tumor, regular imaging analyses did not indicate any relapse. At 17 years of age, a tumor developed in the left leg and increased in size. The resected tumor showed a histopathology of pleomorphic sarcoma positive for the TP53 mutation. Despite amputation of the affected leg, the patient died due to multiple metastases at 18 years of age. A literature review revealed that 14 out of 15 reported malignant PMT-MCT tumors occurred in adults, and found no predictive factors for malignant transformation and treatment outcome. Changes in size or number of the tumors along with intact FGF23 levels have been considered as the only sign of malignant transformation. This pediatric case report and literature review indicate the need for prolonged regular monitoring for PMT-MCT..
6. Kenichiro Yahiro, Yoshihiro Matsumoto, Hisakata Yamada, Makoto Endo, Nokitaka Setsu, Toshifumi Fujiwara, Makoto Nakagawa, Atsushi Kimura, Eijiro Shimada, Seiji Okada, Yoshinao Oda, Yasuharu Nakashima, Activation of TLR4 signaling inhibits progression of osteosarcoma by stimulating CD8-positive cytotoxic lymphocytes, Cancer Immunology, Immunotherapy, 10.1007/s00262-020-02508-9, 69, 5, 745-758, 2020.05, Background: Osteosarcoma (OS) is the most common malignant bone tumor and the prognosis of advanced cases is still poor. Recently, there have been several reports suggesting the relationship between innate immunity and OS, but the detailed mechanism is unknown. We demonstrate the relationship between OS and Toll-like receptor 4 (TLR4) which is one of the most important factors in innate immunity. Methods: We established a syngenic mouse tumor model using C3H/HeN, C3H/HeJ mouse and a highly metastatic OS cell line, LM8. TLR4 activation with lipopolysaccharide (LPS) was performed on both mice and its influence on the progression of OS was evaluated. We also performed CD8 + cells depletion to examine the influence on TLR4 activation effects. Results: Tumor volume of C3H/HeN mice was significantly smaller and overall survival of C3H/HeN mice was significantly longer than C3H/HeJ mice. We found more CD8+ cells infiltrating in lung metastases of C3H/HeN mice and depletion of CD8+ cells canceled the antitumor effects of LPS. Conclusion: TLR4 activation by LPS increased CD8+ cells infiltrating into lung metastases and suppressed OS progression in the mouse model. TLR4 activation may suppress the progression of OS via stimulating CD8+ cells and can be expected as a novel treatment for OS..
7. Kenichiro Yahiro, Yoshihiro Matsumoto, Jun-Ichi Fukushi, Kenichi Kawaguchi, Makoto Endo, Nokitaka Setsu, Keiichiro Iida, Suguru Fukushima, Makoto Nakagawa, Atsushi Kimura, Yoshinao Oda, Yasuharu Nakashima, Class III β-Tubulin Overexpression Induces Chemoresistance to Eribulin in a Leiomyosarcoma Cell Line, Analytical cellular pathology (Amsterdam), 10.1155/2018/8987568, 2018, 2018.01, Eribulin is a new drug to treat soft tissue sarcoma (STS) that exerts antitumor activity by binding to microtubules. The prognosis of STS is poor, and eribulin is expected to improve the treatment outcome. We observed several cases that exhibited resistance to eribulin and developed an eribulin-resistant leiomyosarcoma cell line to investigate the mechanism of resistance. The IC50 of eribulin was 125 times higher in the resistant cell line than in the parental cell line, and eribulin did not induce G2/M arrest in resistant cells. The resistant cell line showed increased expression of MDR1 transcript, but protein levels and functional analysis results were similar to the parental cell line. We found that class III β-tubulin (TUBB3) was overexpressed in the resistant cell line, and siRNA knockdown of TUBB3 partially recovered sensitivity to eribulin. TUBB3 expression in clinical samples varied, suggesting that TUBB3 has the potential to be a biomarker for selection of anticancer drugs and may be a target for overcoming resistance to eribulin..
8. Fusako Kito, Rieko Oyama, Marimu Sakumoto, Mami Takahashi, Kumiko Shiozawa, Zhiwei Qiao, Hiromi Sakamoto, Takeshi Hirose, Nokitaka Setsu, Akihiko Yoshida, Akira Kawai, Tadashi Kondo, Establishment and characterization of novel patient-derived osteosarcoma xenograft and cell line, In Vitro Cellular and Developmental Biology - Animal, 10.1007/s11626-018-0274-2, 54, 7, 528-536, 2018.08, Osteosarcoma is an aggressive mesenchymal malignancy of the bone. Patient-derived models are essential tools for elucidating the molecular mechanisms associated with poor prognosis and the development of novel anticancer drugs. This study described the establishment of a patient-derived cancer model of osteosarcoma. Primary osteosarcoma tumor tissues were obtained from an osteosarcoma patient and inoculated in the skin of immunodeficient mice, followed by transplantation to other mice upon growth. Cells were maintained in monolayer cultures, and the capability of spheroid formation was assessed by seeding the cells on culture dishes. The invasion ability of cells was monitored by Matrigel assay, and genomic and proteomic backgrounds were examined by mass spectrometry. A cell line was established from patient-derived tumors and showed similar histology to that of the primary tumor tissue. Additionally, these cells formed spheroids on low-attachment tissue-culture dishes and exhibited invasive capabilities, and we confirmed that the genomic backgrounds were similar between patient-derived xenograft tumors and the cell line. Furthermore, the proteome of the patient-derived tumors and the cells exhibited similar, but not identical, patterns to that of the original tumor tissue. Our results indicated that this patient-derived xenograft model and cell line would be useful resources for osteosarcoma research..
9. Makoto Endo, Yoshinao Oda, Katsumi Harimaya, Sadafumi Tamiya, Hidetaka Yamamoto, Kenichi Kouhashi, Shuichi Kurihara, Nokitaka Setsu, Suguru Matsuura, Hiroshi Matono, Shuichi Matsuda, Yukihide Iwamoto, Masazumi Tsuneyoshi, Low-grade dedifferentiated liposarcoma of the neck
Magnetic resonance imaging and pathological correlation, Journal of Orthopaedic Science, 10.1007/s00776-009-1407-y, 15, 1, 148-152, 2010.01.
10. Xiaofeng Yan, Masakazu Takahara, Lining Xie, Chisato Gondo, Nokitaka Setsu, Yoshinao Oda, Satoshi Takeuchi, Yating Tu, Yoichi Moroi, Masutaka Furue, Arginine metabolism in soft tissue sarcoma, Journal of Dermatological Science, 10.1016/j.jdermsci.2010.12.009, 61, 3, 211-215, 2011.03, Background: l-Arginine (l-Arg) is a conditionally essential amino acid for humans, which is the substrate for both arginase (ARG) and the inducible form of nitric oxide synthase (iNOS) enzymes. Whether l-Arg metabolism has detrimental or beneficial influence on the tumor growth depends on local up regulation of the NOS or ARG pathways at the tumor site. Objective: To evaluate the expression profile of ARG and iNOS in various histological subtypes of soft tissue sarcomas (STSs). Methods: A series of 81 adult STSs were tested for ARG1, ARG2 and iNOS expression by immunohistochemical analysis. Results: ARG1, ARG2 and iNOS expression was found in tumor cells of all cases of STSs except dermatofibrosarcoma protuberans (DFSP) in a cytoplasmic pattern. However, there was no significant correlation found between ARG, iNOS expression and histopathological parameters. Conversely, the majority of DFSP were devoid of ARG and iNOS expression, while only two cases showed focal and weak expression. Conclusions: Overexpression of l-Arg-metabolizing enzymes ARG and iNOS in tumor cells of all of the STS cases except DFSP may have a role in mediating the biological processes which characterize STSs. New knowledge of the regulation of arginine metabolism in tumor tissues is key to designing sound therapeutic means to effectively prevent tumorigenesis. Further studies are needed to clarify the absence of ARG and iNOS staining in DFSP..
11. Makoto Endo, Chikashi Kobayashi, Nokitaka Setsu, Yusuke Takahashi, Kenichi Kouhashi, Hidetaka Yamamoto, Sadafumi Tamiya, Shuichi Matsuda, Yukihide Iwamoto, Masazumi Tsuneyoshi, Yoshinao Oda, Prognostic significance of p14ARF, p15INK4b, and p16INK4a inactivation in malignant peripheral nerve sheath tumors, Clinical Cancer Research, 10.1158/1078-0432.CCR-10-2393, 17, 11, 3771-3782, 2011.06, Purpose: p14ARF, p15INK4b, and p16INK4a are tumor suppressor genes that are located closely at 9p21 and are often coinactivated by genetic or epigenetic alterations. Malignant peripheral nerve sheath tumor (MPNST) is a rare sarcoma with poor prognosis. However, the prognostic implications of inactivation of p14ARF, p15 INK4b, and p16INK4a in MPNSTs have not been adequately investigated. Here we carried out a genetic, epigenetic, and expression analysis of p14ARF, p15INK4b, and p16INK4a, and clarified the prognostic significance of their inactivation in MPNSTs. Experimental Design: p14ARF, p15INK4b, and p16 INK4a protein expressions were assessed by immunohistochemistry in 129 formalin-fixed samples of MPNST including 85 primary tumors. Thirty-nine samples, for which frozen material was available, were also investigated by Western blotting and quantitative reverse transcription PCR (RT-PCR) to detect p14ARF, p15INK4b, and p16INK4a protein and mRNA expression, and by multiplex real-time PCR, PCR single strand conformation polymorphism and methylation-specific PCR to detect p14ARF, p15 INK4b, and p16INK4a gene alterations. Results: Immunohistochemically decreased expressions of p14ARF, p15 INK4b, and p16INK4a were observed in 48%, 54%, and 49% of primary MPNSTs, respectively, and were significantly correlated with their concordant mRNA levels. As for gene alterations, homozygous deletion of CDKN2A was detected in one third of the cases. Inactivation of p14ARF and p16INK4a was associated with poor prognosis by both univariate and multivariate analyses. Furthermore, cases with inactivation of all p14 ARF, p15INK4b, and p16INK4a genes showed the worst prognosis in a combined prognostic assessment. Conclusion: A comprehensive analysis of p14ARF, p15INK4b, and p16INK4a inactivation status provides useful prognostic information in MPNSTs..
12. Toshifumi Fujiwara, Jun-Ichi Fukushi, Shunsaku Yamamoto, Yoshihiro Matsumoto, Nokitaka Setsu, Yoshinao Oda, Hisakata Yamada, Seiji Okada, Kosuke Watari, Mayumi Ono, Michihiko Kuwano, Satoshi Kamura, Keiichiro Iida, Yuko Okada, Mihoko Koga, Yukihide Iwamoto, Macrophage infiltration predicts a poor prognosis for human ewing sarcoma, American Journal of Pathology, 10.1016/j.ajpath.2011.05.034, 179, 3, 1157-1170, 2011.09, Ewing sarcomaprimitive neuroectodermal tumor (EWS) is associated with the most unfavorable prognosis of all primary musculoskeletal tumors. The objective of the present study was to investigate whether tumor-associated macrophages (TAMs) affect the development of EWS. TAMs were isolated from mouse xenografts using CD11b magnetic beads and examined for their cytokine expression and osteoclastic differentiation. To evaluate the role of TAMs in xenograft formation, liposome-encapsulated clodronate was used to deplete TAMs in mice. Macrophage infiltration and tumor microvascular density were histologically evaluated in 41 patients with EWS, and association with prognosis was examined using Kaplan-Meier survival analysis. In mouse EWS xenografts, TAMs expressed higher concentrations of cytokines including interleukin-6, keratinocyte-derived chemokine, and monocyte chemotactic protein-1. TAMs were more capable than normal monocytes of differentiating into tartrate-resistant acid phosphatasepositive giant cells. Depleting macrophages using liposome-encapsulated clodronate significantly inhibited development of EWS xenografts. In human EWS samples, higher levels of CD68-positive macrophages were associated with poorer overall survival. In addition, enhanced vascularity, increase in the amount of C-reactive protein, and higher white blood cell counts were also associated with poor prognosis and macrophage infiltration. TAMs seem to enhance the progression of EWS by stimulating both angiogenesis and osteoclastogenesis. Further investigation of the behavior of TAMs may lead to development of biologically targeted therapies for EWS..
13. Nokitaka Setsu, Hidetaka Yamamoto, Kenichi Kouhashi, Makoto Endo, Shuichi Matsuda, Ryohei Yokoyama, Kenichi Nishiyama, Yukihide Iwamoto, Yoh Dobashi, Yoshinao Oda, The Akt/mammalian target of rapamycin pathway is activated and associated with adverse prognosis in soft tissue leiomyosarcomas, Cancer, 10.1002/cncr.26448, 118, 6, 1637-1648, 2012.03, BACKGROUND: The Akt/mammalian target of rapamycin (mTOR) pathway mediates cell survival and proliferation and contributes to tumor progression. Soft tissue leiomyosarcoma continues to show poor prognosis, and little is known about its mechanisms of tumor progression. Here the authors investigated the significance of activation of the Akt/mTOR pathway in soft tissue leiomyosarcomas. METHODS: The phosphorylation status of Akt, mTOR, S6, and the eukaryotic translation initiation factor 4E-binding protein (4E-BP1) and the protein expression of phosphatase and tensin homologue (PTEN) were assessed by immunohistochemistry in 145 formalin-fixed paraffin-embedded samples of soft tissue leiomyosarcoma including 129 primary tumors. The expression of phosphorylated Akt and mTOR in comparison with their total forms was assessed by Western blot analysis in 13 frozen samples, which were paired with normal tissue samples. Moreover, 39 frozen tumor samples were analyzed for PIK3CA and AKT1 gene mutation. RESULTS: Immunohistochemically, phosphorylated forms of Akt, mTOR, S6, and 4E-BP1 were positive in 78.3%, 72.6%, 74.5%, and 70.5% of the samples, respectively. These results were correlated with each other, and associated with higher mitotic activity and adverse prognosis. Decreased expression of PTEN was recognized in only 19.7% and had no statistically significant correlation with Akt or other molecules. Immunoblotting showed a high degree of Akt and mTOR phosphorylation in tumor samples compared with that in non-neoplastic tissue. Mutational analysis failed to reveal any PIK3CA or AKT1 mutations around the hot spots. CONCLUSIONS: The Akt/mTOR pathway was activated in most cases of soft tissue leiomyosarcoma and associated with worse clinical behavior and aggressive pathological findings..
14. Nokitaka Setsu, Kenichi Kouhashi, Makoto Endo, Hidetaka Yamamoto, Yoshihiro Ohishi, Kazunobu Sueyoshi, Yukihide Iwamoto, Masazumi Tsuneyoshi, Toru Motoi, Arisa Kumagai, Yoshinao Oda, Inhibin-α and synaptophysin immunoreactivity in synovial sarcoma with granular cell features, Human Pathology, 10.1016/j.humpath.2011.07.012, 43, 6, 850-857, 2012.06, We recognized immunoreactivity for the α subset of inhibin and synaptophysin in synovial sarcomas with granular cell features. Histologic findings of 90 cases of synovial sarcoma were reviewed. Two (2.2%) of the 90 cases had granular cell features, showing sheet or nested proliferation of characteristic epithelioid cells with abundant eosinophilic and granular cytoplasm, in addition to the typical spindle cell component. The 2 cases were both female (aged 86 and 76 years). The tumors were located in the foot and the retroperitoneum and measured 3.5 and 14 cm in maximum diameter. Reverse transcriptase polymerase chain reaction analysis revealed SS18-SSX1 transcripts in both cases. SS18 gene rearrangement was detected in granular cells as well as spindle cells by chromogenic in situ hybridization. Immunohistochemistry found the granular cells to be positive for inhibin-α in both cases and for synaptophysin in 1 case, whereas spindle cells were not. Thirty-six cases (20 monophasic fibrous, 11 biphasic, and 5 poorly differentiated synovial sarcomas) were additionally examined for comparison; they showed no immunoreactivity for inhibin-α or synaptophysin. This is the first report of immunoreactivity for inhibin-α and synaptophysin in synovial sarcoma. These immunohistochemical findings might be characteristic of synovial sarcomas with granular cell features..
15. Makoto Endo, Hidetaka Yamamoto, Nokitaka Setsu, Kenichi Kouhashi, Yusuke Takahashi, Takeaki Ishii, Keiichiro Iida, Yoshihiro Matsumoto, Michiyuki Hakozaki, Mikiko Aoki, Hiroshi Iwasaki, Yoh Dobashi, Kenichi Nishiyama, Yukihide Iwamoto, Yoshinao Oda, Prognostic significance of AKT/mTOR and MAPK pathways and antitumor effect of mTOR inhibitor in NF1-related and sporadic malignant peripheral nerve sheath tumors, Clinical Cancer Research, 10.1158/1078-0432.CCR-12-1067, 19, 2, 450-461, 2013.01, Purpose: Malignant peripheral nerve sheath tumor (MPNST) is a rare soft tissue sarcoma with poor prognosis. MPNSTs occur frequently in patients with neurofibromatosis type 1 (NF1), in which NF1 gene deficiency leads to Ras hyperactivation. Ras activation causes the subsequent activation of the AKT/mTOR and Raf/MEK/ERK pathways and regulates cellular functions. However, the activation profiles of the AKT/ mTORandMAPKpathways in MPNSTs are poorly understood. The purposes of this study are to examine the correlation between the activation of these pathways and clinicopathologic or prognostic factors and to identify candidate target molecules in MPNST. Moreover, we assessed the antitumor effects of the inhibitor of candidate target. Experimental Design: Immunohistochemistry was conducted to evaluate the activation profiles of AKT/ mTOR and MAPK pathways using 135 tumor specimens. Immunohistochemical expressions were confirmed by Western blotting. Then, an in vitro study was conducted to examine the antitumor effect of the mTOR inhibitor on MPNST cell lines. Results: Phosphorylated-AKT (p-AKT), p-mTOR, p-S6RP, p-p70S6K, p-4E-BP1, p-MEK1/2, and p-ERK1/ 2 expressions were positive in 58.2%, 47.3%, 53.8%, 57.1%, 62.6%, 93.4%, and 81.3% of primary MPNSTs, respectively. Positivity for each factor showed no difference between NF1-related and sporadic MPNSTs. Univariate prognostic analysis revealed that p-AKT, p-mTOR, and p-S6RP expressions were associated with poor prognosis. Furthermore, activation of each p-mTOR and p-S6RP was an independent poor prognostic factor by multivariate analysis. mTOR inhibition by Everolimus showed antitumor activity on MPNST cell lines in vitro. Conclusion: mTOR inhibition is a potential treatment option for both NF1-related and sporadic MPNSTs..
16. Nokitaka Setsu, Kenichi Kouhashi, Makoto Endo, Hidetaka Yamamoto, Sadafumi Tamiya, Yusuke Takahashi, Yuichi Yamada, Takeaki Ishii, Shuichi Matsuda, Ryohei Yokoyama, Yukihide Iwamoto, Yoshinao Oda, Phosphorylation of signal transducer and activator of transcription 3 in soft tissue leiomyosarcoma is associated with a better prognosis, International Journal of Cancer, 10.1002/ijc.27655, 132, 1, 109-115, 2013.01, Signal transducer and activator of transcription (STAT) 3 mediates a broad range of biological processes, including cell survival and proliferation, and STAT3 has generally been regarded as a pro-oncogenic transcription factor. We investigated the phosphorylation status of STAT3 and the protein expression of the suppressor of cytokine signaling 3 (SOCS3) by immunohistochemistry in 145 formalin-fixed, paraffin-embedded samples of soft tissue leiomyosarcoma (LMS), including 129 primary tumors. Eight benign soft tissue smooth muscle tumors were also examined. Thirteen frozen LMS samples, which were paired with normal tissue samples, were assessed by Western blot analysis for the phosphorylation of STAT3 and SOCS3 expression. Immunohistochemical study showed that the phosphorylation of STAT3 was not a major event in LMS (37%). Moreover, phosphorylated STAT3 (pSTAT3) expression was significantly correlated with a better prognosis. Overexpression of SOCS3 was recognized in 52% of the cases and negatively correlated with pSTAT3 expression. Among the benign tumors, 63 and 25% were positive for pSTAT3 and SOCS3, respectively. Immunoblotting detected pSTAT3 in all tumor samples, but at lower levels than in non-neoplastic tissue. SOCS3 was detected in 92% (12 out of 13) of tumor tissues, but in none of the normal tissues. Contrary to the previous investigations of many other malignant tumors, STAT3 was inactivated in most LMS cases, likely owing to SOCS3 overexpression. STAT3 might not contribute to the progression of soft tissue LMS, and the phosphorylation status of STAT3 has the potential to be a favorable prognostic marker of LMS. Whats new? Signal transducer and activator of transcription (STAT) 3 mediates a broad range of biological processes and is regarded as pro-oncogenic. Contrary to previous investigations of malignant tumors, STAT3 was found here to be inactivated in most soft tissue leiomyosarcomas, likely due to overexpression of the potential STAT3 inhibitor, suppressor of cytokine signaling (SOCS) 3. Since phosphorylation leads to STAT3 activation, the transcription factors phosphorylation status could serve as a prognostic marker of soft tissue leiomyosarcoma..
17. Suguru Matsuura, Takeaki Ishii, Makoto Endo, Yusuke Takahashi, Nokitaka Setsu, Hidetaka Yamamoto, Sadafumi Tamiya, Yukihide Iwamoto, Yoshinao Oda, Epithelial and cartilaginous differentiation in clear cell chondrosarcoma, Human Pathology, 10.1016/j.humpath.2012.05.012, 44, 2, 237-243, 2013.02, Clear cell chondrosarcoma is a rare cartilaginous bone tumor, and little is known about its pathology. We investigated the immunohistochemical expression profiles of cytokeratins (CAM5.2, AE1/AE3, CK7, CK8, CK18, and CK20), epithelial membrane antigen, SRY (sex-determining region Y)-box 9, type II collagen, runt-related transcription factor 2, and osteocalcin in clear cell chondrosarcoma and compared them with those in chondroblastoma, conventional chondrosarcoma, and osteosarcoma. Of 5 cases of clear cell chondrosarcoma, 3 demonstrated positive staining for AE1/AE3 and some form of cytokeratin in the clear cell component. Of the 5 cases, 4 strongly expressed SRY (sex-determining region Y)-box 9 in the clear cell component but weakly expressed it in the cartilaginous component. Of the 5 cases of clear cell chondrosarcoma, 3 expressed runt-related transcription factor 2 in both the clear cell and cartilaginous components, but no expression of osteocalcin was detected. In chondroblastoma, 8 of 13 cases expressed AE1/AE3, and other cytokeratins, such as CK7 (4/13), CK8 (6/13), CK18 (8/13), and CK20 (3/13), demonstrated a similar staining extensity pattern between the cellular and cartilaginous components. Clear cell chondrosarcoma and chondroblastoma have similar immunohistochemical features in that they both express epithelial and chondrogenetic markers. On the other hand, tumor cells of clear cell chondrosarcoma have no osteoblastic immunohistochemical expression in comparison with chondroblastoma..
18. Y. Fujiwara-Okada, Yoshihiro Matsumoto, Jun-Ichi Fukushi, Nokitaka Setsu, S. Matsuura, S. Kamura, Toshifumi Fujiwara, Keiichiro Iida, M. Hatano, A. Nabeshima, H. Yamada, M. Ono, Yoshinao Oda, Y. Iwamoto, Y-box binding protein-1 regulates cell proliferation and is associated with clinical outcomes of osteosarcoma, British Journal of Cancer, 10.1038/bjc.2012.579, 108, 4, 836-847, 2013.03, Background:Prognosis of osteosarcoma (OS) with distant metastasis and local recurrence is still poor. Y-box binding protein-1 (YB-1) is a multifunctional protein that can act as a regulator of transcription and translation and its high expression of YB-1 protein was observed in OS, however, the role of YB-1 in OS remains unclear.Methods:Y-box binding protein-1 expression in OS cells was inhibited by specific small interfering RNAs to YB-1 (si-YB-1). The effects of si-YB-1 in cell proliferation and cell cycle transition in OS cells were analysed in vitro and in vivo. The association of nuclear expression of YB-1 and clinical prognosis was also investigated by immunohistochemistry.Results: Proliferation of OS cell was suppressed by si-YB-1 in vivo and in vitro. The expression of cyclin D1 and cyclin A were also decreased by si-YB-1. In addition, si-YB-1 induced G1/S arrest with decreased cyclin D1 and cyclin A in OS cell lines. Direct binding of YB-1 in OS cell lines was also observed. Finally, the nuclear expression of YB-1 was significantly related to the poorer overall survival in OS patients.Conclusion:Y-box binding protein-1 would regulate cell cycle progression at G1/S and tumour growth in human OS cells in vitro and in vivo. Nuclear expression of YB-1 was closely associated with the prognosis of OS, thus, YB-1 simultaneously could be a potent molecular target and prognostic biomarker for OS..
19. Yusuke Takahashi, Yoshinao Oda, Hidetaka Yamamoto, Takeaki Ishii, Nokitaka Setsu, Makoto Endo, Shuichi Matsuda, Yukihide Iwamoto, Fibrocartilaginous mesenchymoma arising in the pubic bone
A case report, Pathology International, 10.1111/pin.12052, 63, 4, 226-229, 2013.04, Fibrocartilaginous mesenchymoma (FCM) of the bone is a rare tumor, with only 21 reported cases since 1984. It usually occurs in the long bones of children and adolescents, but in this case, the tumor arose in the pubic bone. The pathological diagnosis of FCM can be challenging, and the treatment should be a wide resection because of its locally aggressive behavior. Histologically, our tumor was composed of a lobular proliferation of spindle cells juxtaposed to the cartilaginous tissue, lacking cytological atypia. Enchondral ossification was observed at the periphery of the cartilaginous nodules, and hypertrophic chondrocytes was recognized, reminiscent of an epiphyseal plate. Intralesional resection followed by phenol and ethanol cauterization was performed in place of the usual wide resection. We report a case of FCM arising in a rare anatomical site, the pubic bone, successfully treated by intralesional resection. One year after the surgery, the patient is free of disease..
20. Makoto Endo, Tatsuya Yoshida, Hidetaka Yamamoto, Takeaki Ishii, Nokitaka Setsu, Kenichi Kouhashi, Tomoya Matsunobu, Yukihide Iwamoto, Yoshinao Oda, Low-grade central osteosarcoma arising from bone infarct, Human Pathology, 10.1016/j.humpath.2012.11.011, 44, 6, 1184-1189, 2013.06, Summary Bone infarct-associated sarcoma is a rare sarcoma, accounting for less than 1% of all bone sarcomas. Its histology usually reflects a high-grade sarcoma, such as malignant fibrous histiocytoma of bone or conventional osteosarcoma. Low-grade sarcoma arising from bone infarct has not been described well in the literature. Here, we present a 17-year follow-up of a female patient with bone infarct in her right humerus, from which a low-grade central osteosarcoma developed during follow-up. A histologic diagnosis of low-grade central osteosarcoma was confirmed by immunohistochemical expression of MDM2 and CDK4. She underwent a wide resection surgery. As of 4 years after surgery, she has remained free of any evidence of recurrence or metastasis. Here, we present clinical and pathologic findings of our case in detail and discuss the differential diagnoses of this extremely rare condition..
21. Hidetaka Yamamoto, Mizuki Handa, Taro Tobo, Nokitaka Setsu, Kohei Fujita, Yumi Oshiro, Yumi Mihara, Yasuji Yoshikawa, Yoshinao Oda, Clinicopathological features of primary leiomyosarcoma of the gastrointestinal tract following recognition of gastrointestinal stromal tumours, Histopathology, 10.1111/his.12159, 63, 2, 194-207, 2013.08, Aims: We aimed to elucidate the clinicopathological and immunohistochemical features of leiomyosarcoma (LMS) of the gastrointestinal (GI) tract. Methods and results: We encountered seven cases of GI-LMS in the colon (n = 4), rectum (n = 1), jejunum (n = 1) and stomach (n = 1). They ranged from 1 to 25 cm (median, 8.5 cm) in size and had high mitotic counts (median 38 per 50 high-power fields). Morphologically, the tumours were composed mainly of spindle cells with eosinophilic cytoplasm and various degrees of nuclear atypia and pleomorphism. Immunohistochemically, the tumours were positive for α-smooth muscle actin (86%), muscle-specific actin (71%), desmin (86%), calponin (71%), h-caldesmon (57%) and smoothelin (71%). All were negative for KIT, CD34, protein kinase C theta and DOG1. Local recurrence and distant metastasis occurred in one and three patients, respectively. We then reviewed 55 cases of GI-LMS from the era following the recognition of gastrointestinal stromal tumours. Among 29 of 55 cases for whom follow-up information was available, the estimated 5-year overall survival rate was 51.6%; tumour size ≥5 cm was correlated significantly with shorter overall survival time (P = 0.0016), while mitotic count (≥50 or ≥100 per 50 high-power fields) proved to be no prognostic factor. Conclusions: GI-LMSs have distinctive clinicopathological and immunohistochemical features and exhibit aggressive biological behaviour..
22. Nokitaka Setsu, Kenichi Kouhashi, Fumiyoshi Fushimi, Makoto Endo, Hidetaka Yamamoto, Yusuke Takahashi, Yuichi Yamada, Takeaki Ishii, Koichirou Yokoyama, Yukihide Iwamoto, Yoshinao Oda, Prognostic impact of the activation status of the Akt/mTOR pathway in synovial sarcoma, Cancer, 10.1002/cncr.28255, 119, 19, 3504-3513, 2013.10, BACKGROUND The Akt/mammalian target of rapamycin (mTOR) pathway, downstream from phosphatidylinositol 3-kinase (PI3K), mediates cell survival and proliferation. Although this pathway reportedly contributes to the progression of synovial sarcoma, its prognostic impact has not been clarified. METHODS The authors analyzed clinicopathologic data and phosphorylation status of Akt (a serine/threonine kinase also known as protein kinase B), mTOR, the eukaryotic translation initiation factor 4E binding protein (4E-BP1), and the S6 ribosomal protein by immunohistochemical analysis of 120 formalin-fixed, paraffin-embedded samples and by Western blot analysis of 24 frozen samples from 112 patients with synovial sarcoma. RESULTS Akt, mTOR, 4E-BP1, and S6 were activated in 76.5%, 67.6%, 59.6%, and 42.6% of samples, respectively. Immunohistochemically positive phosphorylated (p) mTOR (pmTOR) and p4E-BP1 results were correlated with higher mitotic activity, and positive p4E-BP1 results were correlated with greater necrosis. No mutations around the hot spots in the PI3K catalytic subunit α (PI3KCA) and Akt1 genes were observed. In multivariate analysis of clinicopathologic parameters, frequent mitosis was a risk factor for shorter overall survival; and male sex, visceral location, larger tumor size, and frequent mitosis were identified as risk factors for shorter event-free survival. Positive pmTOR and p4E-BP1 results were correlated significantly with shorter overall survival, and positive p4E-BP1 results were correlated with shorter event-free survival in univariate analysis. Positive pAkt results were associated significantly with shorter event-free survival in multivariate analysis. CONCLUSIONS In this study, the Akt/mTOR pathway was activated and was associated with worse clinical and pathologic behavior in patients with synovial sarcoma. The authors propose that this pathway may have potential as a therapeutic target..
23. Makoto Endo, Hidetaka Yamamoto, Katsumi Harimaya, Kenichi Kouhashi, Takeaki Ishii, Nokitaka Setsu, Yukihide Iwamoto, Yoshinao Oda, Conventional spindle cell-type malignant peripheral nerve sheath tumor arising in a sporadic schwannoma, Human Pathology, 10.1016/j.humpath.2013.05.021, 44, 12, 2845-2848, 2013.12, Malignant peripheral nerve sheath tumor is a malignant tumor showing nerve sheath differentiation. Approximately one-half of malignant peripheral nerve sheath tumors arise from a benign peripheral nerve sheath tumor, which is commonly a neurofibroma in patients with neurofibromatosis type 1. Malignant peripheral nerve sheath tumor arising in a sporadic schwannoma of soft tissue is extremely rare. In this condition, malignant cells usually show epithelioid morphology, meeting the diagnostic criteria for epithelioid malignant peripheral nerve sheath tumor. Here, we present an extraordinary case of spindle cell-type malignant peripheral nerve sheath tumor arising in a schwannoma on the back of a 58-year-old woman without neurofibromatosis. The malignant component showed hypercellular spindle cell proliferation with high mitotic activities; in contrast, the benign component showed hypocellular spindle cell proliferation in a palisading pattern and with Verocay bodies. Immunohistochemical S-100 protein staining showed a clear contrast between the malignant (negative) and benign (positive) components, which was useful for differentiating cellular schwannoma. Recognizing this rare condition is helpful in the pathologic diagnosis of schwannoma showing cellular proliferation in part..
24. Nokitaka Setsu, Akihiko Yoshida, Fumiaki Takahashi, Hirokazu Chuman, Ryoji Kushima, Histological analysis suggests an invasion-independent metastatic mechanism in alveolar soft part sarcoma, Human Pathology, 10.1016/j.humpath.2013.07.045, 45, 1, 137-142, 2014.01, Summary Alveolar soft part sarcoma (ASPS) is a rare soft tissue tumor characterized by pseudoalveolar growths associated with abundant sinusoidal vessels. It has a high proclivity to blood-borne metastasis, but the exact mechanism of spread has not been widely discussed, and detailed histological analysis of vascular involvement is still lacking. In this study, we histologically analyzed 32 surgically resected ASPSs, with particular attention to the mode of vascular involvement. Among 188 instances of unequivocal vascular involvement, 184 (98%) were in the form of variously sized cohesive clusters that were completely enveloped by endothelial cells, confirmed by CD31 immunostaining. Discohesive intravascular tumor cells without endothelial wrapping were rare (2%). The clinical relevance of vascular involvement was supported by survival analysis where the average number of vascular involvements per slide was an independent risk factor for shorter progression-free survival. Our findings suggest that ASPSs do not actively break through the vascular walls to initiate the metastatic process. They instead suggest that ASPSs almost exclusively follow the recently postulated "invasion-independent mechanism" for entry into circulation, in which cancer cells are shed into vessels, while being completely enveloped by endothelial cells, and are subsequently entrapped at recipient organs. Because the latter mechanism is reportedly dependent on tumor angiogenesis and vascular remodeling, our data provide a morphological rationale for the use of anti-angiogenic therapy to treat ASPSs..
25. Yusuke Nishida, Eisuke Kobayashi, Daisuke Kubota, Nokitaka Setsu, Koichi Ogura, Yoshikazu Tanzawa, Fumihiko Nakatani, Yoshiharu Kato, Hirokazu Chuman, Akira Kawai, Chronic expanding hematoma with a significantly high fluorodeoxyglucose uptake on 18F-fluorodeoxyglucose positron emission tomography, mimicking a malignant soft tissue tumor
A case report, Journal of Medical Case Reports, 10.1186/1752-1947-8-349, 8, 1, 2014.01, Introduction: Chronic expanding hematoma is a rare persistent hematoma that can sometimes be misdiagnosed as a malignant tumor due to its clinical and radiological features. Case presentation: A 42-year-old Japanese man with a large mass in his leg, suggestive of malignancy, presented to our hospital. He had been aware of the leg swelling for the last eight years. A magnetic resonance imaging scan demonstrated a large mass with two components. One was a large, well-defined cystic mass (13×9cm) showing high intensity on T1-and T2-weighted images, and the other was a solid mass (3.5×2.5cm, adjacent to the large mass) with high intensity on T1-weighted images. Two-[18F]fluoro-2 deoxy-D glucose positron emission tomography images revealed increased uptake with a maximum standardized uptake value of 15.8 in the solid mass. As these findings were considered suggestive of hematoma associated with a malignant lesion, an open biopsy was performed. A pathological examination demonstrated a hematoma with xanthogranuloma, and no malignant cells were evident. Therefore, we resected the tumor including both components, and the histological diagnosis was chronic expanding hematoma. Clinical diagnosis based on 2-[18F]fluoro-2 deoxy-D glucose uptake is sometimes limited by the fact that 2-[18F]fluoro-2 deoxy-D glucose is taken up by not only malignant tumor cells but also macrophages and tissues with granulation or inflammation. Conclusions: Significantly increased standardized uptake value in the peripheral rim of the lesion on 2-[18F]fluoro-2 deoxy-D glucose positron emission tomography imaging, mimicking a soft tissue sarcoma, should be recognized as a potential diagnostic pitfall in cases of chronic expanding hematoma..
26. Yusuke Takahashi, Kenichi Kouhashi, Yuichi Yamada, Makoto Endo, Nokitaka Setsu, Takeaki Ishii, Hidetaka Yamamoto, Yukihide Iwamoto, Yoshinao Oda, Activation of the Akt/mammalian target of rapamycin pathway in myxofibrosarcomas, Human Pathology, 10.1016/j.humpath.2013.12.012, 45, 5, 984-993, 2014.01, The Akt/mammalian target of rapamycin (mTOR) pathway plays important roles in modulating cellular function in response to extracellular signals such as growth factors and cytokines. The Akt/mTOR signaling pathway is activated in certain kinds of sarcomas. Myxofibrosarcoma is a soft tissue sarcoma, characterized by abundant myxoid stroma and frequent local recurrence. Here, we conducted a large-scale examination of the clinicopathological and activation statuses of the Akt/mTOR pathways in myxofibrosarcoma. The phosphorylation status of Akt, mTOR, S6 ribosomal protein, and the eukaryotic translation initiation factor 4E-binding protein, and mitogen-activated protein kinase were assessed by immunohistochemistry in 101 formalin-fixed, paraffin-embedded samples, including 68 primary tumors in myxofibrosarcoma. Immunohistochemical expressions were confirmed by Western blotting with 20 frozen samples, which were paired with normal tissue samples. PIK3CA and AKT1 gene mutations were also analyzed using 12 primary tumor frozen samples. Immunohistochemically, phosphorylations of Akt, mTOR, S6 ribosomal protein, 4E-binding protein, and mitogen-activated protein kinase 1/2 were observed in 64.7%, 45.6%, 42.6%, 63.2%, and 64.7% of samples. Phosphorylated Akt/mTOR pathway proteins were correlated with one another and were also correlated with the phosphorylation of these proteins in the concordant recurrent tumors. Immunoblotting showed a high degree of phosphorylation in tumor samples, compared with that in normal tissue samples. Activation of the Akt/mTOR pathway was correlated with histologic grade and tumor progression. Mutational analysis failed to reveal any PIK3CA or AKT1 mutations around the hot spots. Activation of the Akt/mTOR pathway was associated with histologic malignancy and tumor progression in primary and recurrent myxofibrosarcoma..
27. Yuichi Yamada, Kenichi Kouhashi, Fumiyoshi Fushimi, Yusuke Takahashi, Nokitaka Setsu, Makoto Endo, Hidetaka Yamamoto, Shoji Tokunaga, Yukihide Iwamoto, Yoshinao Oda, Activation of the Akt-mTOR pathway and receptor tyrosine kinase in patients with solitary fibrous tumors, Cancer, 10.1002/cncr.28506, 120, 6, 864-876, 2014.03, BACKGROUND Solitary fibrous tumors (SFTs) are soft tissue tumors of intermediate malignancy that rarely metastasize. Although unresectable SFTs are reported to have a poor prognosis, to the authors' knowledge there is currently no effective therapy. Molecular target therapy is a promising approach for patients with unresectable tumors, but knowledge of the molecular biology of SFTs is currently insufficient to support such therapy. The current study investigated the activation of receptor tyrosine kinases (RTKs) and the Akt-mammalian target of rapamycin (Akt-mTOR) pathway in SFTs as therapeutic targets. METHODS The phosphorylation statuses of Akt-mTOR pathway proteins (p-Akt, p-mTOR, phosphorylated 4E-binding protein [p-4EBP1], and phosphorylated S6 ribosomal protein [p-S6RP]) and RTKs (phosphorylated platelet-derived growth factor receptor-α [p-PDGFRα], p-PDGFRβ, p-c-met, and phosphorylated insulin-like growth factor-1 receptor-β [p-IGF-1Rβ]) were assessed by immunohistochemistry in 66 samples of SFTs, and the data were compared with clinicopathological and histopathological findings. The expression of phosphorylated proteins was assessed by Western blot analysis in 6 frozen samples. RESULTS The immunohistochemical results were as follows: p-Akt, 56.0% (nuclear and cytoplasmic staining); p-mTOR, 69.6% (nuclear and cytoplasmic staining); p-4EBP1, 80.3% (nuclear and cytoplasmic staining); p-S6RP, 69.6% (cytoplasmic staining); p-PDGFRα, 39.0% (cytoplasmic staining); p-PDGFRβ, 52.0% (cytoplasmic staining); p-c-met, 37.8% (nuclear staining) and 19.6% (cytoplasmic staining); and p-IGF-1Rβ, 16.6% (nuclear staining). Phosphorylation of the Akt-mTOR pathway proteins was correlated with one another except for p-Akt with S6RP. p-PDGFRβ and p-IGF-1Rβ were correlated with p-Akt. Moreover, significant relationships were noted between disease-free survival or overall survival and the presence of hypoglycemia, necrosis, cystic and myxoid degeneration, and atypical findings. CONCLUSIONS The Akt/mTOR pathway was activated in approximately 50% of the cases of SFTs and was associated with RTKs, which were phosphorylated at different rates. Thus, the Akt-mTOR pathway may be involved in the tumorigenesis of SFTs. Cancer 2014;120:864-876..
28. Eisuke Kobayashi, Taro Koyama, Kazuhiro Kobayashi, Nokitaka Setsu, Makoto Kawashima, Akira Kawai, Reversible hair depigmentation in a Japanese female treated with pazopanib, Journal of Dermatology, 10.1111/1346-8138.12654, 41, 11, 1021-1022, 2014.11.
29. Eisuke Kobayashi, Nokitaka Setsu, Osteosclerosis induced by denosumab, The Lancet, 10.1016/S0140-6736(14)61338-6, 385, 9967, 2015.01.
30. Nokitaka Setsu, An update of classification and new molecular insights - 2013 world health organization classification of tumours of soft tissue and bone, Japanese Journal of Cancer and Chemotherapy, 42, 3, 291-295, 2015.01.
31. Kensaku Yamaga, Eisuke Kobayashi, Daisuke Kubota, Nokitaka Setsu, Yuya Tanaka, Yusuke Minami, Yoshikazu Tanzawa, Fumihiko Nakatani, Akira Kawai, Hirokazu Chuman, Pediatric myositis ossificans mimicking osteosarcoma, Pediatrics International, 10.1111/ped.12672, 57, 5, 996-999, 2015.10, Myositis ossificans (MO) is a rare benign cause of heterotopic bone formation in soft tissue that most commonly affects young adults, typically following trauma. We report the case of an 11-year-old girl who developed MO mimicking osteosarcoma in her right shoulder. Plain radiography and computed tomography showed poorly defined flocculated densities in the soft tissue and a periosteal reaction along the proximal humerus. On magnetic resonance imaging, the mass displayed an ill-defined margin and inhomogeneous signal change. Histologically, the mass had a pseudosarcomatous appearance. Based on these findings, the patient was initially misdiagnosed with osteosarcoma at another hospital. The diagnosis was difficult because the patient was 11 years old and had no trauma history, with atypical radiographic changes and a predilection for the site of origin for osteosarcomas. We finally made the correct diagnosis of MO by carefully reviewing and reflecting on the pathological differences between stages..
32. Nokitaka Setsu, Eisuke Kobayashi, Akira Kawai, Orthopedic Management of Skeletal Metastases, Japanese Journal of Cancer and Chemotherapy, 42, 11, 1350-1354, 2015.11, Recent advances in drug therapy for metastatic bone diseases are changing the indications for surgical treatments. Bone modifying agents, including bisphosphonates and denosumab, have reduced skeletal-related events. Meanwhile, novel anticancer drugs that have been developed, especially molecular-targeted drugs, can improve the prognoses of patients with bone metastases, thereby potentially increasing the number of patients who could receive benefits from surgeries for metastases. A multidisciplinary approach and the early involvement of surgeons are required for the application of adequate orthopedic treatments, including braces, rehabilitation, and surgeries for local tumor control, palliative fixation, and spinal decompression..
33. Nokitaka Setsu, Eisuke Kobayashi, Naofumi Asano, Naoko Yasui, Hiroshi Kawamoto, Akira Kawai, Keisuke Horiuchi, Severe hypercalcemia following denosumab treatment in a juvenile patient, Journal of Bone and Mineral Metabolism, 10.1007/s00774-015-0677-z, 34, 1, 118-122, 2016.01, A 10-year-old boy diagnosed with unresectable giant cell tumor of bone in the sacrum was treated with a bone modifying agent denosumab. Administration of denosumab showed excellent clinical response without any major complications, and the tumor was surgically removed afterwards. However, 4 months after discontinuing denosumab, the patient developed severe hypercalcemia (15.2 mg/dl). There was a sharp surge in the levels of bone resorption markers, indicating that disregulated overt bone resorption after the discontinuation of denosumab led to hypercalcemia. The patient was treated with bisphosphonate and barely recovered from the life-threatening conditions. This case shows that a robust rebound of bone resorption may occur following cessation of denosumab and suggests that hypercalcemia is an underappreciated side effect of denosumab therapy in children..
34. Akira Maekawa, Kenichi Kouhashi, Nokitaka Setsu, Masaaki Kuda, Kunio Iura, Takeaki Ishii, Tomoya Matsunobu, Tetsuya Nakatsura, Yukihide Iwamoto, Yoshinao Oda, Expression of Forkhead box M1 in soft tissue leiomyosarcoma
Clinicopathologic and in vitro study using a newly established cell line, Cancer Science, 10.1111/cas.12846, 107, 1, 95-102, 2016.01, Leiomyosarcoma (LMS) of soft tissue is a sarcoma with smooth-muscle differentiation, and conventional chemotherapy does not improve its outcome. The application of novel antitumor agents and precise prognostication has been demanded. The expression of the protein Forkhead box M1 (FOXM1), a member of the FOX family, is considered an independent predictor of poor survival in many cancers and sarcomas. However, the expression status of FOXM1 in LMS is poorly understood. The purposes of this study were to examine the correlation between the expression of FOXM1 and clinicopathologic or prognostic factors and to clarify the efficacy of FOXM1 target therapy in LMS. We evaluated the immunohistochemical expressions of FOXM1 using 123 LMS tumor specimens. Univariate and multivariate survival analyses revealed that FOXM1 expression was associated with poor prognosis in LMS. An in vitro study was then carried out to examine the antitumor effect of a FOXM1 inhibitor (thiostrepton) and siRNA on a novel LMS cell line, TC616. We also assessed the efficacy of the combined use of doxorubicin and thiostrepton. Thiostrepton showed dose-dependent antitumor activity and TC616 cells treated with the combination of thiostrepton and doxorubicin showed lower proliferation compared to those treated with either drug individually. FOXM1 interruption by siRNA decreased cell proliferation and increased chemosensitivity. In conclusion, FOXM1 has potential to be a therapeutic target for LMS..
35. Nokitaka Setsu, Mototaka Miyake, Susumu Wakai, Fumihiko Nakatani, Eisuke Kobayashi, Hirokazu Chuman, Nobuyoshi Hiraoka, Akira Kawai, Akihiko Yoshida, Primary Retroperitoneal Myxoid Liposarcomas, American Journal of Surgical Pathology, 10.1097/PAS.0000000000000657, 40, 9, 1286-1290, 2016.09, Myxoid liposarcomas (MLSs) are genetically defined by the presence of DDIT3 gene fusions and most commonly arise in the extremities of young adults. Whether MLSs develop primarily in the retroperitoneum is controversial, and a recent retrospective study found no molecularly confirmed examples. Because MLSs tend to metastasize to deep soft tissues, purported examples of primary retroperitoneal lesions might represent distant metastasis, most commonly from extremities. In addition, well-differentiated or dedifferentiated liposarcomas, which are characterized by MDM2 amplifications, may exhibit prominent myxoid changes and mimic MLSs. Here, we document 5 cases of MLSs that originated in the retroperitoneum that were identified through critical clinicopathologic reevaluation. These cases accounted for 2.3% of 214 primary retroperitoneal liposarcomas and 3.2% of 156 MLSs in our database. They occurred in 3 men and 2 women with a median age of 32 years. All tumors were localized to the retroperitoneum at presentation, and no patient developed extra-abdominal recurrences during the clinical course (median, 50 mo). All 5 cases exhibited at least focal classic histologic findings. All harbored DDIT3 gene rearrangements, and none harbored MDM2 amplifications according to fluorescence in situ hybridization. This study demonstrates that primary MLSs can occur in the retroperitoneum, albeit rarely, and can be accurately diagnosed through combined clinicopathologic and molecular analysis..
36. Shimpei Miyamoto, Masahide Fujiki, Nokitaka Setsu, Akira Kawai, Simultaneous reconstruction of the bone and vessels for complex femoral defect, World Journal of Surgical Oncology, 10.1186/s12957-016-1037-8, 14, 1, 2016.11, Background: Several methods have been reported for intercalary reconstruction of femoral defects. Of these, free vascularized fibula grafts (FVFG) are preferred because of their durability, bone-healing potential, and tolerance to infection. If the bone tumor invades the femoral vessels, simultaneous vascular reconstruction also becomes necessary and significant technical hurdles make limb salvage difficult. Case presentation: We present a 10-year-old girl who underwent limb-sparing surgery for a distal femur osteosarcoma. The femoral defect was 15 cm long, and the femoral vessel defect was 10 cm long. The femur was reconstructed with bilateral FVFG, and the femoral vessels were reconstructed with saphenous vein grafts. The grafts survived without vascular compromise, and the affected limb was preserved successfully. Conclusions: Combined use of bilateral FVFG and autologous vein grafts makes limb-sparing surgery for a large osteosarcoma of the femur possible..
37. Yoshihiro Matsumoto, Shingo Baba, Makoto Endo, Nokitaka Setsu, Keiichiro Iida, Jun-Ichi Fukushi, Kenichi Kawaguchi, Seiji Okada, Hirofumi Bekki, Takuro Isoda, Yoshiyuki Kitamura, Hiroshi Honda, Yasuharu Nakashima, Metabolic Tumor Volume by (18)F-FDG PET/CT Can Predict the Clinical Outcome of Primary Malignant Spine/Spinal Tumors, BioMed Research International, 10.1155/2017/8132676, 2017, 8132676, 2017, BACKGROUND AND PURPOSE: Primary malignant spine/spinal tumors (PMSTs) are rare and life-threatening diseases. In this study, we demonstrated the advantage of volume-based (18)F-FDG PET/CT metabolic parameter, metabolic tumor volume (MTV), for assessing the aggressiveness of PMSTs.
MATERIALS AND METHODS: We retrospectively reviewed 27 patients with PMSTs and calculated SUVmax, MTV, and total lesion glycolysis (TLG) to compare their accuracy in predicting progression-free survival (PFS) and overall survival (OS) by receiver operating characteristic (ROC) curve analysis. Univariate and multivariate analyses were used to compare the reliability of the metabolic parameters and various clinical factors.
RESULTS: MTV exhibited greater accuracy than SUVmax or TLG. The cut-off values for PFS and OS derived from the AUC data were MTV 45 ml and 83 ml and TLG 250 SUV⁎ml and 257 SUV⁎ml, respectively. MTV above cut-off value, but not TLG, was identified as significant prognostic factor for PFS by log-lank test (p = 0.04). In addition, MTV was the only significant predictive factors for PFS and OS in the multivariate analysis.
CONCLUSIONS: MTV was a more accurate predictor of PFS and OS in PMSTs compared to TLG or SUVmax and helped decision-making for guiding rational treatment options..
38. Suguru Fukushima, Makoto Endo, Yoshihiro Matsumoto, Jun-Ichi Fukushi, Tomoya Matsunobu, Ken-Ichi Kawaguchi, Nokitaka Setsu, Keiichiro Iida, Nobuhiko Yokoyama, Makoto Nakagawa, Kenichiro Yahiro, Yoshinao Oda, Yukihide Iwamoto, Yasuharu Nakashima, Hypoxia-inducible factor 1 alpha is a poor prognostic factor and potential therapeutic target in malignant peripheral nerve sheath tumor, PLoS One, 10.1371/journal.pone.0178064, 12, 5, e0178064, 2017, BACKGROUND: Malignant peripheral nerve sheath tumor (MPNST) is a rare soft tissue sarcoma with poor prognosis. Hypoxia-inducible factor 1 (HIF-1) plays a crucial role in the cellular response to hypoxia and regulates the expression of multiple genes involved in tumor progression in various cancers. However, the importance of the expression of HIF-1α in MPNSTs is unclear.
METHODS: The expression of HIF-1α was examined immunohistochemically in 82 MPNST specimens. Cell culture assays of human MPNST cells under normoxic and hypoxic conditions were used to evaluate the impact of anti-HIF-1α-specific siRNA inhibition on cell survival. A screening kit was employed to identify small molecules that inhibited HIF-1α.
RESULTS: The nuclear expression of HIF-1α was positive in 75.6% of MPNST samples (62/82 cases). Positivity for HIF-1α was a significant poor prognostic factor both in univariate (P = 0.048) and multivariate (P ≤ 0.0001) analyses. HIF-1α knockdown abrogated MPNST cell growth, inducing apoptosis. Finally, chetomin, an inhibitor of HIF-1α, effectively inhibited the growth of MPNST cells and induced their apoptosis.
CONCLUSION: Inhibition of HIF-1α signaling is a potential treatment option for MPNSTs..
39. Yoshihiro Matsumoto, Makoto Shinoto, Makoto Endo, Nokitaka Setsu, Keiichiro Iida, Jun-Ichi Fukushi, Kenichi Kawaguchi, Seiji Okada, Hirofumi Bekki, Reiko Imai, Tadashi Kamada, Yoshiyuki Shioyama, Yasuharu Nakashima, Evaluation of risk factors for vertebral compression fracture after carbon-ion radiotherapy for primary spinal and paraspinal sarcoma, BioMed Research International, 10.1155/2017/9467402, 2017, 2017.01, Background and Purpose. Carbon-ion radiotherapy (C-ion RT) was effective therapy for inoperable spinal and paraspinal sarcomas. However, a significant adverse event following radiotherapies is vertebral compression fractures (VCFs). In this study, we investigated the incidence of and risk factors for post-C-ion RT VCFs in patients with spinal or paraspinal sarcomas. Material and Methods. Thirty consecutive patients with spinal or paraspinal sarcomas treated with C-ion RT were retrospectively reviewed. Various clinical parameters and the Spinal Instability Neoplastic Score (SINS) were used to evaluate the risk factors for post-C-ion RT VCFs. Results. The overall incidence of VCFs was 23% (median time: 7 months). Patients with VCFs showed a markedly higher SINS score (median value, 9 points) than those without VCF (5 points). The area under the receiver operating characteristic curve for the SINS score was 0.88, and the optimum SINS cut-off score was 8 points. The cumulative incidence of VCFs at 1 year was 9% for patients with a SINS score under 8 points, versus 80% for those with a SINS score of 8 points or higher (p<0.0001). Conclusions. In patients with a SINS score of 8 points or higher, referral to a spine surgeon for stabilization and multidisciplinary discussion is appropriate..
40. Suguru Fukushima, Makoto Endo, Yoshihiro Matsumoto, Jun-Ichi Fukushi, Tomoya Matsunobu, Ken Ichi Kawaguchi, Nokitaka Setsu, Keiichiro Iida, Nobuhiko Yokoyama, Makoto Nakagawa, Kenichiro Yahiro, Yoshinao Oda, Yukihide Iwamoto, Yasuharu Nakashima, Potential therapeutic target in malignant peripheral nerve sheath tumor, PLoS One, 10.1371/journal.pone.0178064, 12, 5, 2017.05, Background Malignant peripheral nerve sheath tumor (MPNST) is a rare soft tissue sarcoma with poor prognosis. Hypoxia-inducible factor 1 (HIF-1) plays a crucial role in the cellular response to hypoxia and regulates the expression of multiple genes involved in tumor progression in various cancers. However, the importance of the expression of HIF-1α in MPNSTs is unclear. Methods The expression of HIF-1α was examined immunohistochemically in 82 MPNST specimens. Cell culture assays of human MPNST cells under normoxic and hypoxic conditions were used to evaluate the impact of anti-HIF-1α.specific siRNA inhibition on cell survival. A screening kit was employed to identify small molecules that inhibited HIF-1α. Results The nuclear expression of HIF-1α was positive in 75.6% of MPNST samples (62/82 cases). Positivity for HIF-1α was a significant poor prognostic factor both in univariate (P = 0.048) and multivariate (P ≤ 0.0001) analyses. HIF-1α knockdown abrogated MPNST cell growth, inducing apoptosis. Finally, chetomin, an inhibitor of HIF-1α, effectively inhibited the growth of MPNST cells and induced their apoptosis. Conclusion Inhibition of HIF-1α signaling is a potential treatment option for MPNSTs..
41. Yuka Hiraki-Hotokebuchi, Yuichi Yamada, Kenichi Kohashi, Hidetaka Yamamoto, Makoto Endo, Nokitaka Setsu, Kuma Yuki, Takamichi Ito, Yukihide Iwamoto, Masutaka Furue, Yoshinao Oda, Alteration of PDGFRβ-Akt-mTOR Pathway Signaling in Fibrosarcomatous Transformation of Dermatofibrosarcoma Protuberans, Human Pathology, 10.1016/j.humpath.2017.07.001, 2017.07, Dermatofibrosarcoma protuberans (DFSP) is a cutaneous mesenchymal tumor of intermediate malignancy and fibroblastic/myofibroblastic differentiation. Fibrosarcomatous (FS) component is a high-grade component of DFSP. The detailed oncogenic difference between DFSP and FS components is not clear. We thus investigated the Akt-mTOR pathway in both components. We used 65 tumor samples obtained from 65 patients. The phosphorylation of Akt-mTOR pathway proteins (Akt, mTOR, 4EBP1, and S6RP) and PDGFRα/β was assessed by immunohistochemical staining, the results of which were confirmed by western blotting. The immunohistochemical results were as follows: in ordinary DFSP components, p-PDGFRα-positive tumors were 41.9% (18/43 cases), p-PDGFRβ 55.8% (24/43 cases), p-Akt 51.2% (22/43 cases), p-mTOR 39.5% (17/43 cases), p-4EBP1 46.5% (20/43 cases), and p-S6RP 41.8% (18/43 cases); in DFSP components of FS-DFSP, 52.6% (10/19 cases), 47.4% (9/19 cases), 52.6% (10/19 cases), 36.8% (7/19 cases), 52.6% (10/19 cases), and 52.6% (10/19 cases); and in FS components, 45.5% (10/22 cases), 36.4% (8/22 cases), 72.7% (16/22 cases), 54.5% (12/22 cases), 72.7% (16/22 cases), and 68.2% (15/22 cases), respectively. There were significant positive correlations of the phosphorylation of most of the Akt-mTOR pathway proteins (p-Akt, p-mTOR, p-4EBP1, and p-S6RP) with each other (p values <.05). Phospho-PDGFRβ was well correlated with the phosphorylation of Akt-mTOR pathway proteins in DFSP components of ordinary and FS-DFSPs, but these correlations were weaker in FS components. This study suggested the association of activation of Akt-mTOR pathway proteins and PDGFR with the progression of DFSP to FS. The Akt-mTOR pathway is thus a potential therapeutic target in imatinib-resistant DFSP/FS..
42. Keiichiro Iida, Yoshihiro Matsumoto, Nokitaka Setsu, Katsumi Harimaya, Kenichi Kawaguchi, Mitsumasa Hayashida, Seiji Okada, Yasuharu Nakashima, The neurological outcome of radiotherapy versus surgery in patients with metastatic spinal cord compression presenting with myelopathy, Archiv fur orthopadische und Unfall-Chirurgie, 10.1007/s00402-017-2817-5, 138, 1, 7-12, 2018.01, Purpose: While radiotherapy is generally an acceptable treatment for metastatic spinal cord compression, surgical intervention is controversial due to the invasiveness and diversity of diseases in the patients being considered. The ideal treatment, therefore, depends on the situation, and the most acute treatment possible is necessary in patients presenting with myelopathy. We compared the neurological outcomes between radiotherapy and surgery in patients with metastatic spinal cord compression presenting with myelopathy. Methods: A total 54 patients with metastatic spinal cord compression presenting with myelopathy treated in our institution between 2006 and 2016 were analyzed retrospectively. Twenty patients were selected by radiotherapy alone (radiation group), and 36 patients were selected by decompression and stabilization surgery with or without radiotherapy (surgery group). The neurological outcomes and complications were compared between the two treatment groups. Results: Seven patients initially in the radiation group underwent surgery because of a substantial decline in their motor strength during radiotherapy. One of the remaining 13 patients (8%) in the radiation group and 30 of the 34 patients (88%) in the surgery group showed improvement in their neurological symptoms (P < 0.01). One patient (8%) in the radiation group and 21 patients (62%) in the surgery group were ambulatory after treatment (P < 0.01). There were no major complications related to radiotherapy, but surgery-related complications occurred in 9 of 34 (26%) patients, and 6 (18%) patients needed reoperation. Conclusions: Surgical decompression and stabilization may be required to improve the neurological function in patients with metastatic spinal cord compression presenting with myelopathy. However, the high rate of complications associated with surgery should be taken into consideration..
43. Yuichi Yamada, Izumi Kinoshita, Kohashi Kenichi, Hidetaka Yamamoto, Takeshi Iwasaki, Hiroshi Otsuka, Masato Yoshimoto, Shin Ishihara, Yu Toda, Yuki Kuma, Nokitaka Setsu, Yuki Koga, Yumi Honda, Takeshi Inoue, Hiroyuki Yanai, Kyoko Yamashita, Ichiro Ito, Mitsuru Takahashi, Shouichi Ohga, Masutaka Furue, Yasuharu Nakashima, Yoshinao Oda, Histopathological and genetic review of phosphaturic mesenchymal tumours, mixed connective tissue variant, Histopathology, 10.1111/his.13377, 72, 3, 460-471, 2018.02, Aims: Phosphaturic mesenchymal tumour, mixed connective tissue variant (PMT-MCT), is a tumour of uncertain differentiation, characterised by ‘smudgy/grungy’ calcification and vitamin D-resistant phosphaturic osteomalacia. Fibroblast growth factor (FGF)23 is recognised as a reliable marker of PMT-MCT, but quantitative evaluation has never been performed. We reviewed cases of tumour-associated osteomalacia or histologically definitive PMT-MCT without osteomalacia using histological, immunohistochemical and genetic methods and evaluated the diagnostic significance of these findings. Methods and results: A total of 19 tumours from 14 cases diagnosed previously as PMT-MCT were retrieved, on which immunohistochemical staining, reverse transcription–polymerase chain reaction (RT–PCR) and fluorescence in-situ hybridisation (FISH) analysis were performed. Histologically, fibrous capsule, calcification and giant cell reaction tended to be observed in soft-tissue PMT-MCT, while PMT-MCT of bone and multiple PMT-MCT showed an infiltrative growth pattern. The immunohistochemical results were as follows: the tumour cells were positive for FGF23 (nine of 12, 75%), FGFR1 (11 of 11, 100%), CD56 (12 of 14, 85.7%) and E26 oncogene homologue (ERG) (5 of 13, 38.4%). The sole malignant tumour was positive for p53. FGF23 mRNA was detected in seven of 14 formalin-fixed paraffin-embedded (FFPE) specimens and all five frozen specimens by RT–PCR. The level of FGF23 mRNA, which was determined by real-time PCR, varied among the phosphaturic cases. Two of 17 tumours were positive for FGFR1 gene rearrangement. Conclusions: It was considered that PMT-MCT is a histopathological entity with or without phosphaturia, with varying levels of FGF23 mRNA, and with or without fibronectin 1 (FN1)–FGFR1 fusion gene. The authors propose that the histology of PMT-MCT differs depending on its location, such as bone or soft tissue, which could complicate the differential diagnosis..