Targeting sarcoma stem cells using immunological memory T cells.

• Project/Area Number: 20H03807
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 56020:Orthopedics-related
• Research Institution: Sapporo Medical University
• Principal Investigator: TSUKAHARA Tomohide 札幌医科大学, 医学部, 准教授 (20404634)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥17,940,000 (Direct Cost: ¥13,800,000、Indirect Cost: ¥4,140,000); Fiscal Year 2022: ¥3,510,000 (Direct Cost: ¥2,700,000、Indirect Cost: ¥810,000); Fiscal Year 2021: ¥5,720,000 (Direct Cost: ¥4,400,000、Indirect Cost: ¥1,320,000); Fiscal Year 2020: ¥8,710,000 (Direct Cost: ¥6,700,000、Indirect Cost: ¥2,010,000)
• Keywords: CAR-T / TCR-T / HLA/ペプチド複合体 / がん抗原 / 遺伝子改変T細胞 / 抗体 / 癌幹細胞 / 骨肉腫 / 肉腫 / 抗原 / TCR / 免疫 / 癌抗原 / T細胞受容体 / 肉腫幹細胞 / 遺伝子導入T細胞

Outline of Research at the Start

I. 新しいテクノロジーを用いた肉腫幹細胞抗原の同定：肉腫幹細胞は造腫瘍能と抗がん剤耐性を示す．自家細胞傷害性T細胞(CTL)に認識される肉腫幹細胞抗原を新規cDNAディスプレイペプチドライブラリにより同定し，肉腫幹細胞を標的とした免疫療法開発につなげる．
II. TCR導入免疫記憶幹細胞による新しい養子免疫療法の開発：免疫記憶幹細胞は長期生存と分化能を持ち，ナイーブに近い新しいT細胞サブセットである．癌幹細胞抗原特異的なT細胞受容体(TCR)をT細胞に導入・増幅し，さらにNotchシグナル刺激で脱分化させ，疲弊の少ない長期生存が可能なTCR導入免疫記憶幹細胞を大量誘導する．

Outline of Final Research Achievements

We developed a new CAR using B10 scFv clone directed to HLA-A24/cancer stem cell antigen DNAJB8-derived peptide complex (B10 CAR). B10 CAR-T cells showed anti-tumor effects against HLA-A24+/DNAJB8+ cancer and sarcoma cell lines in vitro and in vivo. Moreover, we developed a new TCR using TCR of CTL clone directed to osteosarcoma antigen PBF-derived peptide (PBF TCR). PBF TCR-T cells showed anti-tumor effects against HLA-A24+/PBF+ sarcoma cell lines in vitro and in vivo.

Academic Significance and Societal Importance of the Research Achievements

骨肉腫をはじめとする高い悪性形質を示す肉腫は時に若年者に発症し，肺転移例や化学療法不応例の予後は不良である．本研究成果は，末期肉腫患者に対する遺伝子改変T細胞療法の開発に有用な可能性がある．

Research Products

• [Journal Article] Development of CAR-T cells specifically targeting cancer stem cell antigen DNAJB8 against solid tumours. (2023)
  • Author(s): 127.Watanabe Y, Tsukahara T, Murata K, Hamada S, Kubo T, Kanaseki T, Hirohashi Y, Emori M, Teramoto A, Nakatsugawa M, Yamashita T, Torigoe T
  • Journal Title: Br J Cancer. Volume: 128 Issue: 5 Pages: 886-895
  • DOI: 10.1038/s41416-022-02100-1
  • Peer Reviewed
• [Journal Article] Spatiotemporal metabolic dynamics of the photosensitizer talaporfin sodium in carcinoma and sarcoma. (2021)
  • Author(s): Saito T, Tsukahara T (corresponding author), Suzuki T, Nojima I, Tadano H, Kawai N, Kubo T, Hirohashi Y, Kanaseki T, Torigoe T, Li L.
  • Journal Title: Cancer Sci Volume: 112 Issue: 2 Pages: 550-562
  • DOI: 10.1111/cas.14735
  • Peer Reviewed / Open Access
• [Journal Article] Identification of characteristic subepithelial surface granulomatosis in immune‐related adverse event‐associated enterocolitis (2021)
  • Author(s): Kubo Terufumi、Hirohashi Yoshihiko、Keira Yoshiko、Akimoto Mayuko、Ikeda Tatsuru、Kikuchi Noriaki、Iwaki Hiroyuki、Kikuchi Tomoki、Obata Masahiko、Morita Rena、Kasai Kiyoshi、Segawa Keiko、Tsukahara Tomohide、Kanaseki Takayuki、Murata Kenji、Kikuchi Yasuhiro、Shinkawa Tomoyo、Hasegawa Tadashi、Torigoe Toshihiko
  • Journal Title: Cancer Science Volume: 112 Issue: 3 Pages: 1320-1325
  • DOI: 10.1111/cas.14773
  • Peer Reviewed / Open Access
• [Journal Article] Epithelioid granulomatous lesions express abundant programmed death ligand-1 (PD-L1): a discussion of adverse events in anti-PD-1 antibody-based cancer immunotherapy. (2021)
  • Author(s): Kubo T, Hirohashi Y, Tsukahara T, Kanaseki T, Murata K, Hasegawa T, Torigoe T.
  • Journal Title: Hum Vaccin Immunother. Volume: 18 Issue: 7 Pages: 1-3
  • DOI: 10.1080/21645515.2020.1870364
  • Peer Reviewed
• [Journal Article] Neuregulin-1-β1 and γ-secretase play a critical role in sphere-formation and cell survival of urothelial carcinoma cancer stem-like cells (2021)
  • Author(s): Matsuki M, Inoue R, Murai A, Kubo T, Hashimoto S, Murata K, Kanaseki T, Tsukahara T, Nishida S, Tanaka T, Kitamura H, Masumori N, Hirohashi Y, Torigoe T.
  • Journal Title: Biochem Biophys Res Commun Volume: 552 Pages: 128-135
  • DOI: 10.1016/j.bbrc.2021.03.038
  • Peer Reviewed
• [Journal Article] IL‐13 modulates ΔNp63 levels causing altered expression of barrier‐ and inflammation‐related molecules in human keratinocytes: A possible explanation for chronicity of atopic dermatitis (2021)
  • Author(s): Terufumi Kubo, Sayuri Sato, Tokimasa Hida, Tomoyuki Minowa, Yoshihiko Hirohashi, Tomohide Tsukahara, Takayuki Kanaseki, Kenji Murata, Hisashi Uhara ,Toshihiko Torigoe
  • Journal Title: Immunity, Inflammation and Disease Volume: - Issue: 3 Pages: 734-745
  • DOI: 10.1002/iid3.427
  • Peer Reviewed / Open Access
• [Journal Article] Possible Pseudo-progression of Non-small Cell Lung Carcinoma in a Patient With Clinical Hyper-progression Associated With Trousseau Syndrome Who Was Treated With Pembrolizumab. (2021)
  • Author(s): Shionoya Y, Hirohashi Y, Takahashi H, Hashimoto M, Nishiyama K, Takakuwa Y, Nakatsugawa M, Kubo T, Kanaseki T, Tsukahara T, Torigoe T.
  • Journal Title: Anticancer Res. Volume: 41 Issue: 7 Pages: 3699-3706
  • DOI: 10.21873/anticanres.15161
  • Peer Reviewed
• [Journal Article] Peptide vaccinations elicited strong immune responses that were reboosted by anti-PD1 therapy in a patient with myxofibrosarcoma (2020)
  • Author(s): Tsukahara, T. Watanabe, K. Murata, K. Takahashi, A. Mizushima, E. Shibayama, Y. Kameshima, H. Hatae, R. Ohno, Y. Kawahara, R. Murai, A. Nakatsugawa, M. Kubo, T. Kanaseki, T. Hirohashi, Y. Terui, T. Asanuma, H. Hasegawa, T. Sato, N. Torigoe, T.
  • Journal Title: Cancer immunology immunotherapy Volume: 69 Issue: 2 Pages: 189-197
  • DOI: 10.1007/s00262-019-02455-0
  • Peer Reviewed
• [Journal Article] Development of an artificial antibody specific for HLA/peptide complex derived from cancer stem-like cell/cancer-initiating cell antigen DNAJB8. (2020)
  • Author(s): Tadano H, Tsukahara T (Corresponding author), Mizushima E, Akamatsu A, Watanabe K, Nojima I, Kubo1 T, Kanaseki T, Hirohashi Y, Sato N, Torigoe T.
  • Journal Title: Br J Cancer Volume: 123 Issue: 9 Pages: 1387-1394
  • DOI: 10.1038/s41416-020-1017-1
  • Peer Reviewed
• [Journal Article] Osteosarcoma‐initiating cells show high aerobic glycolysis and attenuation of oxidative phosphorylation mediated by LIN28B (2019)
  • Author(s): Mizushima Emi、Tsukahara Tomohide、Emori Makoto、Murata Kenji、Akamatsu Asuka、Shibayama Yuji、Hamada Shuto、Watanabe Yuto、Kaya Mitsunori、Hirohashi Yoshihiko、Kanaseki Takayuki、Nakatsugawa Munehide、Kubo Terufumi、Yamashita Toshihiko、Sato Noriyuki、Torigoe Toshihiko
  • Journal Title: Cancer Science Volume: 111 Issue: 1 Pages: 36-46
  • DOI: 10.1111/cas.14229
  • Peer Reviewed / Open Access
• [Presentation] がん幹細胞抗原DNAJB8を標的としたCAR-T細胞療法の開発 (2022)
  • Author(s): 渡部裕人，塚原智英，濱田修人，村田憲治，中津川宗秀，鳥越俊彦
  • Organizer: 第81回日本癌学会
• [Presentation] がん幹細胞抗原DNAJB8を標的としたCAR-T細胞療法の開発 (2022)
  • Author(s): 渡部裕人，塚原智英，濱田修人，赤松あすか，村田憲治，中津川宗秀，鳥越俊彦，山下敏彦
  • Organizer: 第37回日本整形外科学会基礎学術集会
• [Presentation] がん幹細胞抗原DNAJB8を標的としたCAR-T細胞療法の開発 (2022)
  • Author(s): 渡部裕人，塚原智英，濱田修人，村田憲治，中津川宗秀，鳥越俊彦
  • Organizer: 第141回北海道整形災害外科学会
• [Presentation] がんと免疫（Human Immunology Priming Seminar ~様々な領域の分子標的療法からヒト免疫を理解しよう~） (2021)
  • Author(s): 塚原智英
  • Organizer: 日本臨床免疫学会
  • Invited
• [Presentation] がんとT細胞免疫（ビギナーズセミナー） (2021)
  • Author(s): 塚原智英
  • Organizer: 日本臨床免疫学会
  • Invited
• [Presentation] 肉腫のImmunopathobiology (2020)
  • Author(s): 塚原智英
  • Organizer: 第24回日本がん免疫学会総会