Kyushu University Academic Staff Educational and Research Activities Database
List of Papers
TAKUJI YAMAUCHI Last modified date:2021.08.12

Assistant Professor / Hematology, Oncology & Cardiovascular medicine / Kyushu University Hospital

1. Suzuki K, Yanagihara T, Matsumoto K, Kusaba H, Yamauchi T, Ikematsu Y, Tanaka K, Otsubo K, Inoue H, Yoneshima Y, Iwama E, Arimura-Omori M, Harada E, Hamada N, Okamoto I, Nakanishi Y., Immune-checkpoint profiles for T cells in bronchoalveolar lavage fluid of patients with immune-checkpoint inhibitor-related interstitial lung disease. Int Immunol., Int Immunol., 10.1093/intimm/dxaa022., 28, 32(8), 547-557, 2020.07.
2. Fumiaki Jinnouchi, Takuji Yamauchi, Ayano Yurino, Takuya Nunomura, Michitaka Nakano, Chika Iwamoto, Teppei Obara, Kohta Miyawaki, Yoshikane Kikushige, Koji Kato, Takahiro Maeda, Toshihiro Miyamoto, Eishi Baba, Koichi Akashi, Katsuto Takenaka, A human SIRPA knock-in xenograft mouse model to study human hematopoietic and cancer stem cells, Blood, 2020.03.
3. Shinichi Kotani, Akinori Yoda, Ayana Kon, Keisuke Kataoka, Yotaro Ochi, Yusuke Shiozawa, Cassandra Hirsch, June Takeda, Hiroo Ueno, Tetsuichi Yoshizato, Kenichi Yoshida, Masahiro M. Nakagawa, Yasuhito Nannya, Nobuyuki Kakiuchi, Takuji Yamauchi, Kosuke Aoki, Yuichi Shiraishi, Satoru Miyano, Takahiro Maeda, Jaroslaw P. Maciejewski, Akifumi Takaori-Kondo, Seishi Ogawa, Hideki Makishima, Molecular pathogenesis of disease progression in MLL-rearranged AML, Leukemia, 10.1038/s41375-018-0253-3, 33, 3, 612-624, 2019.03, Leukemic relapse is frequently accompanied by progressively aggressive clinical course. To understand the molecular mechanism of leukemic relapse, MLL/AF9-transformed mouse leukemia cells were serially transplanted in C57BL/6 mice (N = 96) by mimicking repeated recurrences, where mutations were monitored by exome sequencing (N = 42). The onset of leukemia was progressively promoted with advanced transplants, during which increasing numbers of somatic mutations were acquired (P < 0.005). Among these, mutations in Ptpn11 (p.G60R) and Braf (p.V637E) corresponded to those identified in human MLL-AML, while recurrent mutations affecting Msn (p.R295C) were observed only in mouse but not in human MLL-AML. Another mutated gene of interest was Gnb2 which was reported to be recurrently mutated in various hematological neoplasms. Gnb2 mutations (p.G77R) were significantly increased in clone size (P = 0.007) and associated with earlier leukemia onset (P = 0.011). GNB2 transcripts were significantly upregulated in human MLL-AML compared to MLL-negative AML (P < 0.05), which was supported by significantly increased Gnb2 transcript induced by MLL/AF9 overexpression (P < 0.001). In in vivo model, both mutation and overexpression of GNB2 caused leukemogenesis, and downregulation of GNB2 expression reduced proliferative potential and survival benefit, suggesting a driver role of GNB2. In conclusion, alterations of driver genes over time may play an important role in the progression of MLL-AML..
4. Takuji Yamauchi, Takeshi Masuda, Matthew C. Canver, Michael Seiler, Yuichiro Semba, Mohammad Shboul, Mohammed Al-Raqad, Manami Maeda, Vivien A. C. Schoonenberg, Mitchel A. Cole, Claudio Macias Trevino, Yuichi Ishikawa, Qiuming Yao, Michitaka Nakano, Fumio Arai, Stuart H. Orkin, Bruno Reversade, Silvia Buonamici, Luca Pinello, Koichi Akashi, Daniel E. Bauer and Takahiro Maeda, Genome-wide CRISPR-Cas9 screen identifies leukemia-specific dependence on a pre-mRNA metabolic pathway regulated by the DCPS decapping enzyme , Cancer Cell, 2018.03.
5. Takuji Yamauchi, Takeshi Masuda, Matthew C. Canver, Michael Seiler, Yuichiro Semba, Mohammad Shboul, Mohammed Al-Raqad, Manami Maeda, Vivien A.C. Schoonenberg, Mitchel A. Cole, Claudio Macias-Trevino, Yuichi Ishikawa, Qiuming Yao, Michitaka Nakano, Fumio Arai, Stuart H. Orkin, Bruno Reversade, Silvia Buonamici, Luca Pinello, Koichi Akashi, Daniel E. Bauer, Takahiro Maeda, Genome-wide CRISPR-Cas9 Screen Identifies Leukemia-Specific Dependence on a Pre-mRNA Metabolic Pathway Regulated by DCPS, Cancer Cell, 10.1016/j.ccell.2018.01.012, 33, 3, 386-400.e5, 2018.03, To identify novel targets for acute myeloid leukemia (AML) therapy, we performed genome-wide CRISPR-Cas9 screening using AML cell lines, followed by a second screen in vivo. Here, we show that the mRNA decapping enzyme scavenger (DCPS) gene is essential for AML cell survival. The DCPS enzyme interacted with components of pre-mRNA metabolic pathways, including spliceosomes, as revealed by mass spectrometry. RG3039, a DCPS inhibitor originally developed to treat spinal muscular atrophy, exhibited anti-leukemic activity via inducing pre-mRNA mis-splicing. Humans harboring germline biallelic DCPS loss-of-function mutations do not exhibit aberrant hematologic phenotypes, indicating that DCPS is dispensable for human hematopoiesis. Our findings shed light on a pre-mRNA metabolic pathway and identify DCPS as a target for AML therapy. Yamauchi et al. perform in vitro and in vivo CRISPR-Cas9 genetic screening of p53 WT AML to identify potential therapeutic targets. They find that AML relies on the DCPS decapping enzyme, and a DCPS inhibitor shows anti-leukemia activity in tumor models without impacting normal hematopoiesis..
6. J. Luis Espinoza, Mahmoud I. Elbadry, Masafumi Taniwaki, Kenichi Harada, Ly Quoc Trung, Noriharu Nakagawa, Akiyoshi Takami, Ken Ishiyama, Takuji Yamauchi, Katsuto Takenaka, Shinji Nakao, The simultaneous inhibition of the mTOR and MAPK pathways with Gnetin-C induces apoptosis in acute myeloid leukemia, Cancer Letters, 10.1016/j.canlet.2017.04.027, 400, 127-136, 2017.08, Acute myelogenous leukemia (AML) is a clinically heterogeneous disease that is frequently associated with relapse and a poor prognosis. Among the various subtypes, AML with the monosomal karyotype (AML-MK) has an extremely unfavorable prognosis. We performed screening to identify antitumor compounds that are capable of inducing apoptosis in primary leukemia cells harboring the AML-MK karyotype and identified a naturally occurring stilbene, Gnetin-C, with potent anti-tumor activities against AML cells from patients with various cytogenetic abnormalities, including patients with the AML-MK karyotype. Gnetin-C simultaneously inhibits the ERK1/2 and the AKT/mTOR pathways, two signals that are essential for the survival of leukemia cells. A combination of Gnetin-C with low doses of chemotherapeutic drugs led to synergistic anti-tumor effects against AML cells. In an immunodeficient mouse model of human leukemia, Gnetin-C attenuated the formation of leukemia, depleted leukemia cells and improved survival. These findings suggest that Gnetin-C has antitumor activities in AML and supports the therapeutic potential of blocking two different pathways in AML..
7. Ayako Ichikawa, Hiroaki Miyoshi, Takuji Yamauchi, Fumiko Arakawa, Riko Kawano, Hiroko Muta, Yasuo Sugita, Koichi Akashi, Koichi Ohshima, Composite lymphoma of peripheral T-cell lymphoma and Hodgkin lymphoma, mixed cellularity type; pathological and molecular analysis, Pathology International, 10.1111/pin.12515, 67, 4, 194-201, 2017.04, Composite lymphomas (CLs) are defined as two unrelated lymphomas occurring at the same time within the same tissue. The incidence of these tumors is low. Of all possible combinations between lymphomas, the least frequent are the ones combining peripheral T-cell lymphoma (PTCL) and Hodgkin lymphoma (HL). We recently identified five cases of CL composed of PTCL and classical HL, mixed cellularity type. We investigated histological and clinical features of these cases. Immunostaining was performed on paraffin sections. PTCL cells were positive for CD8 and TIA-1 in four of the five cases. Hodgkin and Reed-Sternberg (HRS) cells were positive for CD30 and weakly positive for PAX5 in all cases, positive for CD15 in three of five cases, positive for CD20 in one of five cases, and negative for EBER. Monoclonal rearrangement of the T-cell receptor (TCR) and immunoglobulin heavy chain (IGH) genes was confirmed by polymerase chain reaction (PCR) using whole paraffin sections. We concluded more precisely the monoclonality of the IGH rearrangement of HRS cells based on single-cell PCR for IGH and DNA sequencing analysis after laser microdissection of single cells in one case. HL can occur in CD8-positive and TIA-1-positive PTCL. Clinicians should recognize the possibility of these CL..
8. Taro Tochigi, Takatoshi Aoki, Yoshikane Kikushige, Tomohiko Kamimura, Yoshikiyo Ito, Takahiro Shima, Takuji Yamauchi, Yasuo Mori, Goichi Yoshimoto, Kenjiro Kamezaki, Koji Kato, Katsuto Takenaka, Hiromi Iwasaki, Koichi Akashi, Toshihiro Miyamoto, Mobilization of human immature hematopoietic progenitors through combinatory use of bortezomib and immunomodulatory drugs, International journal of hematology, 10.1007/s12185-016-2148-2, 105, 4, 423-432, 2017.04, Combination use of the proteasome inhibitor bortezomib and the immunomodulatory drugs lenalidomide or thalidomide has provided superior outcomes in multiple myeloma over their single use; however, these combinations can produce significant toxicities. Unexpectedly, we found a small but significant increase in the population of immature granulocytes and erythrocytes/megakaryocytes in peripheral blood in 16 of 22 patients (73%) treated with dexamethasone in combination with bortezomib and immunomodulatory drugs (triplet), but not in any of 25 patients treated with either bortezomib or immunomodulatory drugs with dexamethasone (doublet). These immature cells gradually increased to a peak level (mean 2.6% per white blood cells) with triplet therapy, and disappeared immediately after therapy cessation. The numbers of circulating CD34+ cells and colony-forming cells derived from peripheral blood mononuclear cells increased after triplet therapy compared with those in patients treated by either bortezomib or immunomodulatory drugs plus dexamethasone. Furthermore, triplet regimen downregulated the expression of CXCR4, a chemokine receptor essential for bone marrow retention, on CD34+ cells, suggesting an unexpected effect on normal hematopoietic stem/progenitor cells through the reduced interaction with the bone marrow microenvironment. Our observations suggest that combination use should be carefully evaluated to exert synergistic anti-myeloma effects while avoiding unexpected adverse events..
9. Shinya Daitoku, Katsuto Takenaka, Takuji Yamauchi, Ayano Yurino, Fumiaki Jinnouchi, Takuya Nunomura, Tetsuya Eto, Tomohiko Kamimura, Masakazu Higuchi, Naoki Harada, Noriyuki Saito, Toshihiro Miyamoto, Hiromi Iwasaki, Koichi Akashi, Calreticulin mutation does not contribute to disease progression in essential thrombocythemia by inhibiting phagocytosis, Experimental Hematology, 10.1016/j.exphem.2016.05.001, 44, 9, 817-825.e3, 2016.09, Somatic mutations of calreticulin (CALR) have been observed in many cases of essential thrombocythemia (ET) or primary myelofibrosis that harbor non-mutated Janus kinase 2 (JAK2). CALR mainly localizes within the endoplasmic reticulum lumen, but a small fraction of the total CALR pool is distributed over the cell surface. Cell surface CALR is known to transduce prophagocytic “eat me” signals to macrophages and acts as one of the important regulators for macrophage engulfment. In this study, we attempted to clarify whether mutant CALR may affect the threshold for macrophage engulfment and play an integral role in the pathogenesis of CALR-mutated ET. First, we compared the surface expression levels of CALR on hematopoietic stem and progenitor cells (HSPCs) and mature blood cells in patients with myeloproliferative neoplasms and found that the surface expression of mutant CALR did not change. Next, we compared the threshold for macrophage phagocytosis of each HSPC fraction and mature blood cells and found no significant change in the efficiency of macrophage engulfment. Our data suggest that CALR mutation does not affect sensitivity to phagocytosis by macrophages. Finally, we analyzed the phosphorylation statuses of molecules downstream of JAK2 at each HSPC level in patients with ET and found that CALR mutations activated the JAK-STAT pathway in a manner similar to that associated with JAK2 mutations. These results indicate that mutant CALR causes myeloproliferation because of the activation of JAK-STAT pathway and not by the inhibition of phagocytosis, which is similar to the myeloproliferation caused by JAK2 V617F mutation..
10. Ayano Yurino, Katsuto Takenaka, Takuji Yamauchi, Takuya Nunomura, Yasufumi Uehara, Fumiaki Jinnouchi, Kohta Miyawaki, Yoshikane Kikushige, Koji Kato, Toshihiro Miyamoto, Hiromi Iwasaki, Yuya Kunisaki, Koichi Akashi, Enhanced Reconstitution of Human Erythropoiesis and Thrombopoiesis in an Immunodeficient Mouse Model with KitWv Mutations, Stem Cell Reports, 10.1016/j.stemcr.2016.07.002, 7, 3, 425-438, 2016.09, In human-to-mouse xenograft models, reconstitution of human hematopoiesis is usually B-lymphoid dominant. Here we show that the introduction of homozygous KitWv mutations into C57BL/6.Rag2nullIl2rgnull mice with NOD-Sirpa (BRGS) strongly promoted human multi-lineage reconstitution. After xenotransplantation of human CD34+CD38 cord blood cells, these newly generated C57BL/6.Rag2nullIl2rgnullNOD-Sirpa KitWv/Wv (BRGSKWv/Wv) mice showed significantly higher levels of human cell chimerism and long-term multi-lineage reconstitution compared with BRGS mice. Strikingly, this mouse displayed a robust reconstitution of human erythropoiesis and thrombopoiesis with terminal maturation in the bone marrow. Furthermore, depletion of host macrophages by clodronate administration resulted in the presence of human erythrocytes and platelets in the circulation. Thus, attenuation of mouse KIT signaling greatly enhances the multi-lineage differentiation of human hematopoietic stem and progenitor cells (HSPCs) in mouse bone marrow, presumably by outcompeting mouse HSPCs to occupy suitable microenvironments. The BRGSKWv/Wv mouse model is a useful tool to study human multi-lineage hematopoiesis..
11. Takeshi Sugio, Koji Kato, Takatoshi Aoki, Takanori Ohta, Noriyuki Saito, Shuro Yoshida, Ichiro Kawano, Hideho Henzan, Masanori Kadowaki, Ken Takase, Tsuyoshi Muta, Kohta Miyawaki, Takuji Yamauchi, Takahiro Shima, Shuichiro Takashima, Yasuo Mori, Goichi Yoshimoto, Kenjiro Kamezaki, Katsuto Takenaka, Hiromi Iwasaki, Ryosuke Ogawa, Yuju Ohno, Tetsuya Eto, Tomohiko Kamimura, Toshihiro Miyamoto, Koichi Akashi, Mogamulizumab Treatment Prior to Allogeneic Hematopoietic Stem Cell Transplantation Induces Severe Acute Graft-versus-Host Disease, Biology of Blood and Marrow Transplantation, 10.1016/j.bbmt.2016.05.017, 22, 9, 1608-1614, 2016.09, Mogamulizumab (MOG), a humanized anti-CC chemokine receptor 4 (CCR4) monoclonal antibody, has recently played an important role in the treatment of adult T cell leukemia/lymphoma (ATLL). Because CCR4 is expressed on normal regulatory T cells as well as on ATLL cells, MOG may accelerate graft-versus-host disease (GVHD) by eradicating regulatory T cells in patients with allogeneic hematopoietic stem cell transplantation (allo-HSCT). However, there is limited information about its safety and efficacy in patients treated with MOG before allo-HSCT. In the present study, 25 patients with ATLL were treated with MOG before allo-HSCT, after which 18 patients (72%) achieved remission. The overall survival and progression-free survival at 1 year post-transplantation were 20.2% (95% CI, 6.0% to 40.3%) and 15.0% (95% CI, 4.3% to 32.0%), respectively. The cumulative incidence of acute GVHD was 64.0% (95% CI, 40.7% to 80.1%) for grade II-IV and 34.7% (95% CI, 15.8% to 54.4%) for grade III-IV. The cumulative incidence of transplantation-related mortality (TRM) was 49.0% (95% CI, 27.0% to 67.8%). Six of 7 patients with acute GVHD grade III-IV died from GVHD, which was the leading cause of death. In particular, a shorter interval from the last administration of MOG to allo-HSCT was associated with more severe GVHD. MOG use before allo-HSCT may decrease the ATLL burden; however, it is associated with an increase in TRM due to severe GVHD. Because MOG is a potent anti-ATLL agent, new treatment protocols should be developed to integrate MOG at suitable doses and timing of administration to minimize unwanted GVHD development..
12. T. Shima, H. Iwasaki, T. Yamauchi, M. Kadowaki, M. Kiyosuke, T. Mochimaru, K. Takenaka, T. Miyamoto, K. Akashi, T. Teshima, Preserved in vivo reconstitution ability of PBSCs cryopreserved for a decade at -80 °c, Bone Marrow Transplantation, 10.1038/bmt.2015.147, 50, 9, 1195-1200, 2015.09, PBSC products for auto- and allografting can be cryopreserved in liquid nitrogen with controlled-rate freezing until their use. Alternatively, they can be stored at -80 °C in a mechanical chest freezer, but it remains to be clarified whether PBSCs can be stored for the long term. We evaluated viability and functions of PBSCs cryopreserved for more than 10 years with this simplified method. Although recovery rate and viability of CD34 + cells were significantly decreased, myeloid differentiation potential and in vivo reconstitution and self-renewal potential of CD34 + cells in a xenogeneic engraftment assay were maintained for more than 10 years. These results indicate that PBSCs can be stored at -80 °C for years. Although accumulation of clinical engraftment data is required to confirm our results, this simplified cryopreservation will thus meet the increasing worldwide demand for PBSC transplantation in a region with limited resources..
13. Chika Iwamoto, Katsuto Takenaka, Shingo Urata, Takuji Yamauchi, Takahiro Shima, Takuro Kuriyama, Shinya Daitoku, Yasuyuki Saito, Toshihiro Miyamoto, Hiromi Iwasaki, Issay Kitabayashi, Katsuhiko Itoh, Junji Kishimoto, Daisuke Kohda, Takashi Matozaki, Koichi Akashi, The BALB/c-specific polymorphic SIRPA enhances its affinity for human CD47, inhibiting phagocytosis against human cells to promote xenogeneic engraftment, Experimental Hematology, 10.1016/j.exphem.2013.11.005, 42, 3, 163-171.e1, 2014.03, It has been shown that in xenotransplantation of human cells into immunodeficient mice, the mouse strain background is critical. For example, the nonobese diabetic (NOD) strain is most efficient, the BALB/c is moderate, and the C57BL/6 is inefficient for human cell engraftment. We have shown that the NOD-specific polymorphism of the signal regulatory protein-alpha (Sirpa) allows NOD SIRPA to bind human CD47, and the resultant "don't eat me" signaling by this binding prevents host macrophages to engulf human grafts, thereby inhibiting rejection. Here we tested whether the efficient xenotransplantation capability of the BALB/c strain is also mediated by the SIRPA-CD47 self-recognition system. BALB/c SIRPA was capable of binding to human CD47 at an intermediate level between those of C57BL/6 SIRPA and NOD SIRPA. Consistent with its binding activity, BALB/c-derived macrophages exhibited a moderate inhibitory effect on human long-term culture-initiating cells in in vitro cultures, and showed moderate phagocytic activity against human hematopoietic stem cells. The increased affinity of BALB/c SIRPA for human CD47 was mounted at least through the BALB/c-specific L29V SNP within the IgV domain. Thus, the mouse strain effect on xenogeneic engraftment might be ascribed mainly to the binding affinity of strain-specific polymorphic SIRPA with human CD47. This information should be useful for developing a novel immunodeficient strain with superior efficiency for xenogeneic transplantation of human cells..
14. T. Shima, N. Forraz, N. Sato, T. Yamauchi, H. Iwasaki, Katsuto Takenaka, K. Akashi, C. Mcguckin, T. Teshima, A novel filtration method for cord blood processing using a polyester fabric filter, International Journal of Laboratory Hematology, 10.1111/ijlh.12039, 35, 4, 436-446, 2013.08, Introduction: Cord blood (CB) is being increasingly used as a source of hematopoietic stem cells for transplantation to treat diseases of the blood and immune systems, and there is an urgent need to expand CB banking worldwide. CB processing requires costly machinery or a clean room that hampers wider application of CBT particularly in the developing countries. Methods: We developed a novel filtration system using a nonchemical-coated and nonwoven polyester fabric filter, which traps cells through affinity and does not require centrifugation or potentially toxic chemicals. Results: Cell processing with the device resulted in minimum cell loss of total cells and CD34+ cells, without impairing the ability of CD34+ cells to engraft and differentiate both in vivo and in vitro. Conclusion: CB processing with this device is simple, cost-effective, and nontoxic without requiring costly equipment will thus facilitate international CB banking, which helps in meeting the increasing worldwide demand for CB for allogeneic hematopoietic stem cell transplantation..
15. Mitsuhiro Fukata, Fumihiko Ishikawa, Yuho Najima, Takuji Yamauchi, Yoriko Saito, Katsuto Takenaka, Kohta Miyawaki, Hideki Shimazu, Kazuya Shimoda, Takaaki Kanemaru, Kei ichiro Nakamura, Keita Odashiro, Koji Nagafuji, Mine Harada, Koichi Akashi, Contribution of Bone Marrow-Derived Hematopoietic Stem/Progenitor Cells to the Generation of Donor-Marker+ Cardiomyocytes In Vivo, PloS one, 10.1371/journal.pone.0062506, 8, 5, 2013.05, Background:Definite identification of the cell types and the mechanism relevant to cardiomyogenesis is essential for effective cardiac regenerative medicine. We aimed to identify the cell populations that can generate cardiomyocytes and to clarify whether generation of donor-marker+ cardiomyocytes requires cell fusion between BM-derived cells and recipient cardiomyocytes.Methodology/Principal Findings:Purified BM stem/progenitor cells from green fluorescence protein (GFP) mice were transplanted into C57BL/6 mice or cyan fluorescence protein (CFP)-transgenic mice. Purified human hematopoietic stem cells (HSCs) from cord blood were transplanted into immune-compromised NOD/SCID/IL2rγnull mice. GFP+ cells in the cardiac tissue were analyzed for the antigenecity of a cardiomyocyte by confocal microscopy following immunofluorescence staining. GFP+ donor-derived cells, GFP+CFP+ fused cells, and CFP+ recipient-derived cells were distinguished by linear unmixing analysis. Hearts of xenogeneic recipients were evaluated for the expression of human cardiomyocyte genes by real-time quantitative polymerase chain reaction. In C57BL/6 recipients, Lin-/lowCD45+ hematopoietic cells generated greater number of GFP+ cardiomyocytes than Lin-/lowCD45- mesenchymal cells (37.0+/-23.9 vs 0.00+/-0.00 GFP+ cardiomyocytes per a recipient, P = 0.0095). The number of transplanted purified HSCs (Lin-/lowSca-1+ or Lin-Sca-1+c-Kit+ or CD34-Lin-Sca-1+c-Kit+) showed correlation to the number of GFP+ cardiomyocytes (P<0.05 in each cell fraction), and the incidence of GFP+ cardiomyocytes per injected cell dose was greatest in CD34-Lin-Sca-1+c-Kit+ recipients. Of the hematopoietic progenitors, total myeloid progenitors generated greater number of GFP+ cardiomyocytes than common lymphoid progenitors (12.8+/-10.7 vs 0.67+/-1.00 GFP+ cardiomyocytes per a recipient, P = 0.0021). In CFP recipients, all GFP+ cardiomyocytes examined coexpressed CFP. Human troponin C and myosin heavy chain 6 transcripts were detected in the cardiac tissue of some of the xenogeneic recipients.Conclusions/Significance:Our results indicate that HSCs resulted in the generation of cardiomyocytes via myeloid intermediates by fusion-dependent mechanism. The use of myeloid derivatives as donor cells could potentially allow more effective cell-based therapy for cardiac repair..
16. Takuji Yamauchi, Katsuto Takenaka, Shingo Urata, Takahiro Shima, Yoshikane Kikushige, Takahito Tokuyama, Chika Iwamoto, Mariko Nishihara, Hiromi Iwasaki, Toshihiro Miyamoto, Nakayuki Honma, Miki Nakao, Takashi Matozaki, Koichi Akashi, Polymorphic Sirpa is the genetic determinant for NOD-based mouse lines to achieve efficient human cell engraftment, Blood, 10.1182/blood-2012-06-440354, 121, 8, 1316-1325, 2013.02, Current mouse lines efficient for human cell xenotransplantation are backcrossed into NOD mice to introduce its multiple immunodeficient phenotypes. Our positional genetic study has located the NOD-specific polymorphic Sirpa as a molecule responsible for its high xenograft efficiency: it recognizes human CD47 and the resultant signaling may cause NOD macrophages not to engulf human grafts. In the present study, we established C57BL/6.Rag2 null//2rgnull mice harboring NOD-Sirpa (BRGS). BRGS mice engrafted human hematopoiesis with an efficiency that was equal to or even better than that of the NOD.Rag1null//2rgnull strain, one of the best xenograft models. Consequently, BRGS mice are free from other NOD-related abnormalities; for example, they have normalized C5 function that enables the evaluation of complement-dependent cytotoxicity of antibodies against human grafts in the humanized mouse model. Our data show that efficient human cell engraftment found in NOD-based models is mounted solely by their polymorphic Sirpa. The simplified BRGS line should be very useful in future studies of human stem cell biology..
17. Takuro Kuriyama, Katsuto Takenaka, Kentaro Kohno, Takuji Yamauchi, Shinya Daitoku, Goichi Yoshimoto, Yoshikane Kikushige, Junji Kishimoto, Yasunobu Abe, Naoki Harada, Toshihiro Miyamoto, Hiromi Iwasaki, Takanori Teshima, Koichi Akashi, Engulfment of hematopoietic stem cells caused by down-regulation of CD47 is critical in the pathogenesis of hemophagocytic lymphohistiocytosis, Blood, 10.1182/blood-2012-02-408864, 120, 19, 4058-4067, 2012.11, Hemophagocytic lymphohistiocytosis (HLH) is characterized by deregulated engulfment of hematopoietic stem cells (HSCs) by BM macrophages, which are activated presumably by systemic inflammatory hypercytokinemia. In the present study, we show that the pathogenesis of HLH involves impairment of the antiphagocytic system operated by an interaction between surface CD47 and signal regulatory protein α (SIRPA). In HLH patients, changes in expression levels and HLH-specific polymorphism of SIRPA were not found. In contrast, the expression of surface CD47 was down-regulated specifically in HSCs in association with exacerbation of HLH, but not in healthy subjects. The number of BM HSCs in HLH patients was reduced to approximately 20% of that of healthy controls and macrophages from normal donors aggressively engulfed HSCs purified from HLH patients, but not those from healthy controls in vitro. Furthermore, in response to inflammatory cytokines, normal HSCs, but not progenitors or mature blood cells, down-regulated CD47 sufficiently to be engulfed by macrophages. The expression of prophagocytic calreticulin was kept suppressed at the HSC stage in both HLH patients and healthy controls, even in the presence of inflammatory cytokines. These data suggest that the CD47-SIRPA antiphagocytic system plays a key role in the maintenance of HSCs and that its disruption by HSC-specific CD47 down-regulation might be critical for HLH development..
18. Yoshikane Kikushige, Fumihiko Ishikawa, Toshihiro Miyamoto, Takahiro Shima, Shingo Urata, Goichi Yoshimoto, Yasuo Mori, Tadafumi Iino, Takuji Yamauchi, Tetsuya Eto, Hiroaki Niiro, Hiromi Iwasaki, Katsuto Takenaka, Koichi Akashi, Self-Renewing Hematopoietic Stem Cell Is the Primary Target in Pathogenesis of Human Chronic Lymphocytic Leukemia, Cancer Cell, 10.1016/j.ccr.2011.06.029, 20, 2, 246-259, 2011.08, We report here that in chronic lymphocytic leukemia (CLL), the propensity to generate clonal B cells has been acquired already at the hematopoietic stem cell (HSC) stage. HSCs purified from patients with CLL displayed lymphoid-lineage gene priming and produced a high number of polyclonal B cell progenitors. Strikingly, their maturation into B cells was restricted always to mono- or oligo-clones with CLL-like phenotype in xenogeneic recipients. These B cell clones were independent of the original CLL clones because they had their own immunoglobulin VDJ genes. Furthermore, they used preferentially VH genes frequently used in human CLL, presumably reflecting the role of B cell receptor signaling in clonal selection. These data suggest that HSCs can be involved in leukemogenesis even in mature lymphoid tumors..
19. Takatoshi Aoki, Toshihiro Miyamoto, Yasuo Mori, Goichi Yoshimoto, Takuji Yamauchi, Kenjiro Kamezaki, Katsuto Takenaka, Hiromi Iwasaki, Naoki Harada, Koji Nagafuji, Nobuyuki Shimono, Takanori Teshima, Koichi Akashi, Successful allogeneic stem cell transplantation in two patients with acute myelogenous leukaemia and invasive aspergillosis by antifungal combination therapy, Mycoses, 10.1111/j.1439-0507.2010.01858.x, 54, 4, e255-e259, 2011.07, Invasive aspergillosis (IA) is an important cause of infectious morbidity and mortality in patients who undergo haematopoietic stem cell transplantation (HSCT). History of IA before allogeneic HSCT is still challenging because of the high risk of recurrence after HSCT. Recent advances in early-stage diagnosis and new, more effective classes of antifungal agents have improved the management of IA in the HSCT recipients. We report two cases with acute myelogenous leukaemia after primary failure of induction chemotherapy with the patients developing pulmonary IA. They responded well to a combination of voriconazole (VCZ) and micafungin, resulting in a remarkable reduction of pulmonary IA lesions at short intervals. Thereafter, antifungal therapy was switched to liposomal amphotericin B (L-AmB), followed by conditioning regimen for allogeneic HSCT, because of the possibility of VCZ altering the metabolism of chemotherapeutic agents and calcineurin inhibitors. Successful engraftment was achieved without severe adverse side-effects or aggravation of IA after HSCT. Combining VCZ with micafungin followed by L-AmB throughout HSCT could be advantageous in stabilising IA in HSCT patients..
20. Seido Oku, Katsuto Takenaka, Takuro Kuriyama, Kotaro Shide, Takashi Kumano, Yoshikane Kikushige, Shingo Urata, Takuji Yamauchi, Chika Iwamoto, Haruko K. Shimoda, Toshihiro Miyamoto, Koji Nagafuji, Junji Kishimoto, Kazuya Shimoda, Koichi Akashi, JAK2 V617F uses distinct signalling pathways to induce cell proliferation and neutrophil activation
Research paper, British Journal of Haematology, 10.1111/j.1365-2141.2010.08249.x, 150, 3, 334-344, 2010.08, Summary The acquired JAK2 V617F mutation is observed in the majority of patients with BCR-ABL1 negative chronic myeloproliferative neoplasms (MPN). BCR-ABL1 negative MPN displays myeloproliferation with an elevated leucocyte alkaline phosphatase (LAP) activity, a neutrophil activation marker. We tried to separate the downstream signalling of JAK2 V617F to stimulate myeloproliferation and LAP activity. NB4, a myeloid lineage cell line, was transduced with Jak2 V617F mutation or wild-type Jak2. We found that Jak2 V617F mutation, but not wild-type Jak2 enhanced LAP expression in NB4-derived neutrophils and proliferation of NB4 cells. JAK2 V617F induces constitutive phosphorylation of STAT3 and STAT5, and uses signalling targets such as Ras/MEK/ERK and PI3K/Akt pathways. By using MEK1/2 inhibitor U0126, PI3K inhibitor LY294002, and STAT3 or STAT5 siRNAs, JAK2 V617F was found to specifically use the STAT3 pathway to enhance LAP expression, while STAT5, Ras/MEK/ERK and PI3K/Akt, but not STAT3 pathways, were able to stimulate cell proliferation. These data strongly suggest that JAK2 V617F uses distinct signalling pathways to induce typical pathological features of MPN, such as high LAP activity and enhanced cell proliferation..
21. T. Aoki, K. Kamezaki, T. Miyamoto, K. Nagafuji, Y. Mori, T. Yamauchi, K. Takenaka, H. Iwasaki, N. Harada, N. Shimono, T. Teshima, K. Akashi, Cord blood stem cell transplantation in a patient with disseminated mucormycosis and acute myelogenous leukemia, Transplant Infectious Disease, 10.1111/j.1399-3062.2010.00496.x, 12, 3, 277-279, 2010.06.
22. Yasuo Mori, Takatoshi Aoki, Katsuto Takenaka, Takuji Yamauchi, Asataro Yamamoto, Kenjiro Kamezaki, Hiromi Iwasaki, Naoki Harada, Toshihiro Miyamoto, Koji Nagafuji, Takanori Teshima, Koichi Akashi, Successful treatment of refractory advanced nasal NK/T cell lymphoma with unrelated cord blood stem cell transplantation incorporating focal irradiation, International journal of hematology, 10.1007/s12185-009-0453-8, 91, 1, 107-111, 2010.01, Nasal natural killer (NK)/T cell lymphoma is a rare disease with a poor prognosis. We report the case of a 52-year-old woman with progressive advanced nasal NK/T cell lymphoma, with local invasiveness and bone marrow involvement, who was successfully treated with unrelated cord blood transplantation (UCBT). The patient was initially refractory to conventional chemotherapy. She was therefore treated with local irradiation, which induced a partial response. The patient then underwent UCBT using a conditioning regimen consisting of cyclophosphamide and total body irradiation. Acute graft-versus-host disease involving the skin was observed, but it was well controlled without systemic administration of corticosteroids. The patient remained in complete remission for 18 months after UCBT. Although the observation period has been relatively short and longer follow-up is needed, our observations suggest that incorporating focal irradiation to conditioning regimen for local control might be an effective treatment option for advanced nasal NK/T cell lymphoma in the setting of UCBT..
23. Yukari Asai, Hiroshi Ouchi, Tukasa Ohosima, Ryuji Nakano, Yujiro Yamano, Ichiro Inoshima, Takuji Yamauchi, Satoshi Fukuyama, Hiromasa Inoue, Yoichi Nakanishi, A case of secondary pulmonary alveolar proteinosis associated with myelodysplastic syndrome, complicated with disseminated M. abscessus infection, Nihon Kokyūki Gakkai zasshi = the journal of the Japanese Respiratory Society, 47, 12, 1120-1125, 2009.12, A 27-year-old man was admitted to our hospital complaining of a persistent high fever since August 2007. Chest radiography showed infiltration shadows in the right lower lung field. Chest CT revealed scattered small nodular shadows and patchy consolidations in the right lower lobe. He was diagnosed as secondary pulmonary alveolar proteinosis (sPAP) associated with myelodysplastic syndrome (MDS), confirmed by video-assisted thoracic surgery (VATS) and bone marrow aspiration. Sera were negative for anti-granulocyte-macrophage colony-stimulating factor (GM-CSF) autoantibody. He developed a subcutaneous abscess and meningitis caused by M. absessus after VATS. After a long-course of antibiotic therapy, an allogenic peripheral blood stem cell transplantation was performed. But he died of graft versus host disease and M. abscessus sepsis 87 days after transplantation..
24. Takatoshi Aoki, Toshihiro Miyamoto, Shuro Yoshida, Asataro Yamamoto, Takuji Yamauchi, Goichi Yoshimoto, Yasuo Mori, Kenjiro Kamezaki, Hiromi Iwasaki, Katsuto Takenaka, Naoki Harada, Koji Nagafuji, Takanori Teshima, Koichi Akashi, Additional acquisition of t(1;21)(p32;q22) in a patient relapsing with acute myelogenous leukemia with NUP98-HOXA9, International journal of hematology, 10.1007/s12185-008-0198-9, 88, 5, 571-574, 2009.12, We report a 29-year-old Japanese male with acute myelogenous leukemia (AML)-M4 with a cryptic t(7;11)(p15;p15), in which a chimeric NUP98-HOXA9 fusion was detected by polymerase chain reaction analysis and a chromosomal analysis showed 46,XY. The patient received intensive chemotherapy and underwent autologous stem cell transplantation, and remission was confirmed by the disappearance of NUP98-HOXA9. However, 6 months after transplantation, the patient relapsed; NUP98-HOXA9 was detected again and karyotypic analysis revealed 46,XY, t(1;21)(p32;q22). Fluorescent in situ hybridization (FISH) analysis using an AML1-ETO translocation dual probe, showed that the 21q22 breakpoint involved AML1 locus. A retrospective FISH analysis showed that t(1;21) was absent at onset. This is the first reported case with AML who had a cryptic t(7;11)(p15;p15), and additionally acquired t(1;21)(p32;q22) at relapse..
25. Takuji Yamauchi, Yasuo Mori, Toshihiro Miyamoto, Kenjiro Kamezaki, Takatoshi Aoki, Asataro Yamamoto, Katsuto Takenaka, Hiromi Iwasaki, Naoki Harada, Koji Nagafuji, Takanori Teshima, Koichi Akashi, Second unrelated cord blood transplantation using a reduced-intensity conditioning regimen combined with gemtuzumab ozogamicin in patients with relapsed acute myelogenous leukemia, International journal of hematology, 10.1007/s12185-009-0405-3, 90, 3, 416-420, 2009.10, Gemtuzumab ozogamicin (GO) is an effective molecular-targeted agent for CD33-positive acute myelogenous leukemia (AML) patients who are resistant to conventional chemotherapy. Recent prospective trials have revealed the safety and efficacy of GO as part of conditioning following allogeneic bone marrow or peripheral blood stem cell transplantation (SCT). We report here for the first time three AML cases that relapsed after allogeneic SCT and underwent unrelated cord blood transplantation (UCBT) following reduced-intensity conditioning (RIC) comprising fludarabine, melphalan, and low-dose total body irradiation combined with GO. Primary neutrophil engraftment occurred in all cases, while recovery of platelet count was delayed. Only one case of reversible hepatic sinusoidal obstruction syndrome was documented. Non-relapse mortality at day 100 was not documented. Notably, one patient who responded to GO survived for 6 months after UCBT in remission with excellent performance status, while the remaining cases relapsed early. These data suggest that GO may be safely combined with RIC for UCBT after previous allogeneic SCT..