Research Experience 【 display / non-display 】

Research Experience 【 display / non-display 】

• 2023.11

  -

  Now

  Sapporo Medical University   医学部附属研究所免疫学研究所分子医学部門/医学部病理学第一講座   特任助教

• 2022.06

  -

  2023.10

  Sapporo Medical University   Research Institute for Frontier Medicine   特任助教

• 2020.04

  -

  2022.05

  Sapporo Medical University   医学部 病理学第一講座   特任助教

• 2016.04

  -

  2020.03

  プリンセスマーガレットがんセンター   平野研究室   博士研究員

Research Areas 【 display / non-display 】

Research Areas 【 display / non-display 】

• Life sciences   Molecular biology  

• Life sciences   Immunology  

• Life sciences   Orthopedics  

Affiliation 【 display / non-display 】

Affiliation 【 display / non-display 】

• Sapporo Medical University   Research Institute for Frontier Medicine   特任助教  

Research Interests 【 display / non-display 】

Research Interests 【 display / non-display 】

• 骨軟部肉腫

• 腫瘍免疫学

• T細胞受容体

• 養子免疫療法

Papers 【 display / non-display 】

Papers 【 display / non-display 】

• Landscape mapping of shared antigenic epitopes and their cognate TCRs of tumor-infiltrating T lymphocytes in melanoma.

  Kenji Murata, Munehide Nakatsugawa, Muhammed A Rahman, Linh T Nguyen, Douglas G Millar, David T Mulder, Kenji Sugata, Hiroshi Saijo, Yukiko Matsunaga, Yuki Kagoya, Tingxi Guo, Mark Anczurowski, Chung-Hsi Wang, Brian D Burt, Dalam Ly, Kayoko Saso, Alexandra Easson, David P Goldstein, Michael Reedijk, Danny Ghazarian, Trevor J Pugh, Marcus O Butler, Tak W Mak, Pamela S Ohashi, Naoto Hirano

  eLife   9  2020.04  [International journal]

  Authorship：   Lead author

  　View Summary

  HLA-restricted T cell responses can induce antitumor effects in cancer patients. Previous human T cell research has largely focused on the few HLA alleles prevalent in a subset of ethnic groups. Here, using a panel of newly developed peptide-exchangeable peptide/HLA multimers and artificial antigen-presenting cells for 25 different class I alleles and greater than 800 peptides, we systematically and comprehensively mapped shared antigenic epitopes recognized by tumor-infiltrating T lymphocytes (TILs) from eight melanoma patients for all their class I alleles. We were able to determine the specificity, on average, of 12.2% of the TILs recognizing a mean of 3.1 shared antigen-derived epitopes across HLA-A, B, and C. Furthermore, we isolated a number of cognate T cell receptor genes with tumor reactivity. Our novel strategy allows for a more complete examination of the immune response and development of novel cancer immunotherapy not limited by HLA allele prevalence or tumor mutation burden.

  DOI PubMed

• Establishment of potent TCR-T cells specific for cisplatin-resistance related tumor-associated antigen, CLSPN using codon-optimization.

  Kanta Hori, Shuhei Yamada, Kenji Murata, Haruka Miyata, Yuka Mizue, Aiko Murai, Tomoyuki Minowa, Kenta Sasaki, Naoki Shijubou, Terufumi Kubo, Rena Morita, Serina Tokita, Takayuki Kanaseki, Tomohide Tsukahara, Takashige Abe, Nobuo Shinohara, Yoshihiko Hirohashi, Toshihiko Torigoe

  Human vaccines & immunotherapeutics   20 ( 1 ) 2414542 - 2414542  2024.12  [International journal]

  　View Summary

  Adoptive T cell therapy, using T cell receptor-engineered T (TCR-T) cells and chimeric antigen receptor T (CAR-T) cells, is a potent immunotherapy option. Bladder cancer is a prevalent urological malignancy, particularly in cases of muscle invasion and metastasis, for which systemic therapy is crucial. Immunotherapy utilizing immune checkpoint blockade has been approved for bladder cancer treatment. The antitumor effect of an immune checkpoint blockade based on cytotoxic T cells (CTLs) and the patient's immune status is essential. The chemotherapeutic drug cisplatin (CDDP) is a key drug in bladder cancer treatment. However, it has been shown to suppress T cells, making combination therapy with CDDP and immunotherapy difficult. To address this, we developed TCR-T cells specific for bladder cancer cells. In previous studies, we found that the tumor-associated antigen CLSPN is overexpressed in CDDP-resistant bladder cancer cells and that the antigenic peptide HLA-A*02:01/CLSPN1254-1262, encoded by CLSPN, could be targeted by a CTL clone. The TCR was cloned from the HLA-A*02:01/CLSPN1254-1262 specific CTL clone yc3. We also designed a codon-optimized TCR sequence using GeneArt® GeneOptimizer® (Opt TCR) and compared the TCR-T cells using the original TCR sequence (Ori TCR-T cells) and the codon-optimized TCR sequence (Opt TCR-T cells). Opt TCR-T cells exhibited higher TCR transduction efficiency, higher TCR expression levels, higher avidity, and greater cytotoxicity than did Ori TCR-T cells. These results suggest that HLA-A*02:01/CLSPN1254-1262 specific Opt TCR-T cells are promising candidates for CDDP combination therapy.

  DOI PubMed

• Identification and phenotypic characterization of neoantigen-specific cytotoxic CD4+ T cells in endometrial cancer

  Minami Fusagawa, Serina Tokita, Kenji Murata, Tasuku Mariya, Mina Umemoto, Shintaro Sugita, Kazuhiko Matsuo, Yoshihiko Hirohashi, Tsuyoshi Saito, Takayuki Kanaseki, Toshihiko Torigoe

  Cancer Immunology Research ( American Association for Cancer Research (AACR) )   2024.12

  　View Summary

  Abstract Tumor-reactive CD4+ T cells often accumulate in the tumor microenvironment (TME) in human cancer, but their functions and roles in antitumor responses remain elusive. Here, we investigated the immunopeptidome of HLA class II–positive (HLA-II+) endometrial cancer with an inflamed TME using a proteogenomic approach. We identified HLA-II neoantigens, one of which induced polyclonal CD4+ tumor-infiltrating lymphocyte (TIL) responses. We then experimentally demonstrated that neoantigen-specific CD4+ TILs lyse target cells in an HLA-II–dependent manner. Single cell transcriptomic analysis of the TME coupled with T-cell receptor (TCR) sequencing revealed the presence of CD4+ T-cell clusters characterized by CXCL13 expression. The CXCL13+ clusters contained two subclusters with distinct cytotoxic gene expression patterns. The identified neoantigen-specific CD4+ T cells were found exclusively in one of the CXCL13+ subclusters characterized by granzyme B and CCL5 expression. These results demonstrate the involvement of tumor-reactive CD4+ T cells with cytotoxic function in immune surveillance of endometrial cancer and reveal their transcriptomic signature.

  DOI

• Single-cell profiling of acral melanoma infiltrating lymphocytes reveals a suppressive tumor microenvironment.

  Tomoyuki Minowa, Kenji Murata, Yuka Mizue, Aiko Murai, Munehide Nakatsugawa, Kenta Sasaki, Serina Tokita, Terufumi Kubo, Takayuki Kanaseki, Tomohide Tsukahara, Toshiya Handa, Sayuri Sato, Kohei Horimoto, Junji Kato, Tokimasa Hida, Yoshihiko Hirohashi, Hisashi Uhara, Toshihiko Torigoe

  Science translational medicine   16 ( 776 ) eadk8832  2024.12  [International journal]

  Authorship：   Corresponding author

  　View Summary

  Acral lentiginous melanoma (ALM) is the most common melanoma subtype in non-Caucasians. Despite advances in cancer immunotherapy, current immune checkpoint inhibitors remain unsatisfactory for ALM. Hence, we conducted comprehensive immune profiling using single-cell phenotyping with reactivity screening of the T cell receptors of tumor-infiltrating T lymphocytes (TILs) in ALM. Compared with cutaneous melanoma, ALM showed a lower frequency of tumor-reactive CD8 clusters and an enrichment of regulatory T cells with direct tumor recognition ability, suggesting a suppressive immune microenvironment in ALM. Tumor-reactive CD8 TILs showed heterogeneous expression of coinhibitory molecules, including KLRC1 (NKG2A), in subpopulations with therapeutic implications. Overall, our study provides a foundation for enhancing the efficacy of immunotherapy in ALM.

  DOI PubMed

• Aldehyde Dehydrogenese-1 High Cancer Stem-like Cells/Cancer-initiating Cells Escape from Cytotoxic T Lymphocytes due to Lower Expression of Human Leukocyte Antigen Class 1.

  Tomohide Shirosaki, Noriko Kawai, Yuma Ebihara, Aiko Murai, Terufumi Kubo, Rena Morita, Kenji Murata, Takayuki Kanaseki, Tomohide Tsukahara, Toshiaki Shichinohe, Yoshihiko Hirohashi, Satoshi Hirano, Toshihiko Torigoe

  Anticancer research   44 ( 5 ) 1877 - 1883  2024.05  [International journal]

  　View Summary

  BACKGROUND/AIM: Human gastric cancer stem-like cells (CSCs)/cancer-initiating cells can be identified as aldehyde dehydrogenase-high (ALDHhigh) cells. Cancer immunotherapy employing immune checkpoint blockade has been approved for advanced gastric cancer cases. However, the effectiveness of cancer immunotherapy against gastric CSCs/CICs remains unclear. This study aimed to investigate the susceptibility of gastric CSCs/CICs to immunotherapy. MATERIALS AND METHODS: Gastric CSCs/CICs were isolated as ALDHhigh cells using the human gastric cancer cell line, MKN-45. ALDHhigh clone cells and ALDHlow clone cells were isolated using the ALDEFLUOR assay. ALDH1A1 expression was assessed via qRT-PCR. Sphere-forming ability was evaluated to confirm the presence of CSCs/CICs. A model neoantigen, AP2S1, was over-expressed in ALDHhigh clone cells and ALDHlow clone cells, and susceptibility to AP2S1-specific TCR-T cells was assessed using IFNγ ELISPOT assay. RESULTS: Three ALDHhigh clone cells were isolated from MKN-45 cells. ALDHhigh clone cells exhibited a stable phenotype in in vitro culture for more than 2 months. The High-36 clone cells demonstrated the highest sphere-forming ability, whereas the Low-8 cells showed the lowest sphere-forming ability. High-36 cells exhibited lower expression of HLA-A24 compared to Low-8 cells. TCR-T cells specific for AP2S1 showed lower reactivity to High-36 cells compared to Low-8 cells. CONCLUSION: High-36 cells and Low-8 cells represent novel gastric CSCs/CICs and non-CSCs/CICs, respectively. ALDHhigh CSCs/CICs evade T cells due to lower expression of HLA class 1.

  DOI PubMed

display all >>

Books and Other Publications 【 display / non-display 】

Books and Other Publications 【 display / non-display 】

• 次世代CAR-T細胞

  村田 憲治, 平野 直人( Part： Contributor, TCR-T細胞療法の現状と期待：適応拡大と有効性向上のための新技術)

  羊土社  2023.11 ISBN: 9784758125734

Misc 【 display / non-display 】

Misc 【 display / non-display 】

• Comprehensive single-cell immune profiling of tumor-infiltrating lymphocytes in acral melanoma

  箕輪智幸, 箕輪智幸, 村田憲治, 廣橋良彦, 宇原久, 鳥越俊彦

  日本病理学会会誌   112 ( 1 )  2023

  J-GLOBAL

• Development of HLA-A2/Claspin peptide-specific antibodies targeting platinum-resistant cancer cells

  水江由佳, 廣橋良彦, 塚原智英, 金関貴幸, 久保輝文, 村田憲治, 鳥越俊彦

  日本病理学会会誌   112 ( 1 )  2023

  J-GLOBAL

• The landscape of immunogenic HLA ligands in human cancer revealed by the proteogenomic approach

  鳥越俊彦, 廣橋良彦, 塚原智英, 塚原智英, 金関貴幸, 久保輝文, 村田憲治, 村田憲治, 中津川宗秀

  日本癌学会学術総会抄録集(Web)   81st  2022

  J-GLOBAL

• Immunopathology research toward the development of next-generation immunotherapy of cancer

  鳥越俊彦, 廣橋良彦, 塚原智英, 金関貴幸, 久保輝文, 村田憲治

  日本病理学会会誌   111 ( 2 )  2022

  J-GLOBAL

• Neoantigen identification and reactive TCR analysis in CRC tissue using a proteogenomic approach

  時田芹奈, 時田芹奈, 金関貴幸, 村田憲治, 中津川宗秀, 平間知美, 平間知美, 佐藤昇志, 佐藤昇志, 秦史壯, 鳥越俊彦

  日本病理学会会誌   111 ( 1 )  2022

  J-GLOBAL

display all >>

Awards 【 display / non-display 】

Awards 【 display / non-display 】

• Research award

  2018   北海道整形災害外科学会   Identification of a novel human T cell population with the characteristics of stem-like chemo-resistance.

Research Projects 【 display / non-display 】

Research Projects 【 display / non-display 】

• 腫瘍反応性TCRをプローブとして用いた新たな肉腫抗原同定法の開発

  日本医療研究開発機構(AMED) 次世代がん医療加速化研究事業(P-PROMOTE)

  Project Year :

  2022

  -

  2023

   

  村田 憲治

• 抗肉腫ヒト人工T細胞療法に向けたTCR・抗原ペア網羅的解析プラットフォーム開発

  若手研究

  Project Year :

  2021.04

  -

  2023.03

   

  村田 憲治

  　View Summary

  骨軟部肉腫は若年者に多く発生する悪性度の高い希少がんであり、化学療法に反応しない進行期肉腫患者の予後は不良である。近年、「がん免疫療法」は飛躍的に進化し、手術療法、抗がん剤治療、放射線治療の3大治療に次ぐ第4の治療法として注目をされており、ステージ 4 のがん患者でも劇的回復を示す症例がみられる。肉腫に対しては、がん精巣抗原の一つである NY-ESO-1 を標的として、T 細胞受容体（TCR）遺伝子を導入した T 細胞(TCR-T 細胞)を用いた T 細胞輸注療法が末期滑膜肉腫の 50%以 上に部分効果を示すことも報告されている。しかしながら、他の難治性肉腫疾患に対する既存の免疫療法の奏効率はいまだ低く、さらに肉腫に発現する特異抗原とそれを認識するTCR遺伝子の報告は少ない。 本研究の目的は、骨軟部肉腫に対する効果的な遺伝子改変T細胞療法の実行化に向けて、肉腫浸潤Tリンパ球（TIL）由来の肉腫反応性TCRが認識する抗原を同定するための新たな解析プラットフォームを開発することである。 平滑筋肉腫2症例（LMS02、LMS07）の切除検体からTILを分離して10x Genomics社のChromium Controllerを用いて単細胞レベルでのTCRレパトアと遺伝子発現解析を行なった。いずれもTCRのクローナリティが高いT細胞が存在し、さらにそれらは活性化マーカーや疲弊マーカーの発現がやや高く、腫瘍を認識している可能性が高い細胞集団であった。CD8陽性で、PD－1陽性かつ4-1BB陽性のTCRをサブクローニングし、TCR-T細胞を作成した。このTCRを抗原探索のプローブとして用い、新たな肉腫抗原を同定する。

• Immune response of tumor-infiltrating T lymphocytes in sarcoma using single-cell analysis

  Grant-in-Aid for Research Activity Start-up

  Project Year :

  2020.09

  -

  2022.03

   

  MURATA Kenji

  　View Summary

  We isolated tumor-infiltrating lymphocytes (TILs) from a myxoinflammatory fibroblastic sarcoma tissue and performed single-cell analyses of the TILs. CD8+ T cells were dominant in the TILs, and top two T-cell receptors (TCRs) were more than 40% of CD8+ T cells. CD8+ TILs of high TCR clonality did not express memory markers, but highly expressed exhaustion markers, activation makers, and cytokine genes, suggesting that these TCRs might recognize tumors.