Development of a novel method for T-cell preparations targeting cancer using genome editing in human and monkey induced pluripotent stem cells

• Project/Area Number: 19K07712
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 50020:Tumor diagnostics and therapeutics-related
• Research Institution: Shiga University of Medical Science
• Principal Investigator: TERADA Koji 滋賀医科大学, 医学部, 准教授 (70342722)
• Project Period (FY): 2019-04-01 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2021: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2020: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2019: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: T細胞受容体 / T細胞 / がん / カニクイザル / ゲノム編集 / iPS細胞 / TIL / TCR / 免疫療法

Outline of Research at the Start

細胞傷害性T細胞は、がん細胞が特異的に発現するがん抗原をＴ細胞受容体で認識し、がん細胞を攻撃する。近年、がん抗原特異的なＴ細胞受容体の遺伝子を、細胞傷害性T細胞に導入するがん免疫細胞療法が開発されているが、安全性や治療効果など、まだ克服すべき問題が存在する。本研究計画では、安全性と治療効果が共に高い細胞傷害性T細胞を効率よく作製する、新しいがん免疫細胞療法の開発を推進する。

Outline of Final Research Achievements

To develop effective adoptive cell transfer therapy using T cell receptor (TCR)-engineered T cells, it is critical to isolate tumor-reactive TCRs that have potent anti-tumor activity. Characterization of tumor reactivity of tumor-infiltrating lymphocytes (TILs) from non-human primate tumors could improve anti-tumor activity of TCR-engineered T cells in preclinical research. In this study, we isolated TCR genes from TILs in a cynomolgus macaque model of tumor transplantation in which the tumors were infiltrated with CD8+ T cells and were eventually rejected. We identified multiple TCR pairs with high frequency. We showed that the recurrent TCRs exhibited cytotoxic activity to tumor cells in vitro and potent anti-tumor activity in mice transplanted with tumor cells. These results imply that this tumor transplantation macaque model recapitulates key features of human TILs and can serve as a platform toward pre- clinical studies of non-human primate tumor models.

Academic Significance and Societal Importance of the Research Achievements

本研究では、カニクイザルの腫瘍から得られた腫瘍浸潤リンパ球（TIL）の細胞表面発現分子とT細胞受容体遺伝子を解析した。その結果、カニクイザルのTILはヒトのTILと共通の性質を有すること、外来性にTCRを発現させたT細胞を用いたがん免疫療法の前臨床研究においてカニクイザルが有用な動物モデルとなる可能性があることなどが示唆された。

Research Products

• [Journal Article] Isolation of TCR genes with tumor-killing activity from tumor-infiltrating and circulating lymphocytes in a tumor rejection cynomolgus macaque model (2022)
  • Author(s): Terada Koji、Kondo Kenta、Ishigaki Hirohito、Nagashima Ayaka、Satooka Hiroki、Nagano Seiji、Masuda Kyoko、Kawamura Teruhisa、Hirata Takako、Ogasawara Kazumasa、Itoh Yasushi、Kawamoto Hiroshi、Agata Yasutoshi
  • Journal Title: Molecular Therapy - Oncolytics Volume: 24 Pages: 77-86
  • DOI: 10.1016/j.omto.2021.12.003
  • NAID: 120007182844
  • Peer Reviewed / Open Access
• [Journal Article] Regeneration of Tumor-Antigen-Specific Cytotoxic T Lymphocytes from iPSCs Transduced with Exogenous TCR Genes (2020)
  • Author(s): Maeda Takuya、Nagano Seiji、Kashima Soki、Terada Koji、Agata Yasutoshi、Ichise Hiroshi、Ohtaka Manami、Nakanishi Mahito、Fujiki Fumihiro、Sugiyama Haruo、Kitawaki Toshio、Kadowaki Norimitsu、Takaori-Kondo Akifumi、Masuda Kyoko、Kawamoto Hiroshi
  • Journal Title: Molecular Therapy - Methods & Clinical Development Volume: 19 Pages: 250-260
  • DOI: 10.1016/j.omtm.2020.09.011
  • NAID: 120007169387
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Characterization of tumour-infiltrating lymphocytes in a tumour rejection cynomolgus macaque model (2020)
  • Author(s): Satooka Hiroki、Ishigaki Hirohito、Todo Kagefumi、Terada Koji、Agata Yasutoshi、Itoh Yasushi、Ogasawara Kazumasa、Hirata Takako
  • Journal Title: Scientific Reports Volume: 10 Issue: 1 Pages: 8414-8414
  • DOI: 10.1038/s41598-020-65488-x
  • NAID: 120006862372
  • Peer Reviewed / Open Access
• [Journal Article] Cytotoxic T lymphocytes regenerated from iPS cells have therapeutic efficacy in a patient-derived xenograft solid tumor model. (2020)
  • Author(s): Kashima S, Maeda T, Masuda K, Nagano S, Inoue T, Takeda M, Kono Y, Kobayashi T, Saito S, Higuchi T, Ichise H, Kobayashi Y, Iwaisako K, Terada K, Agata Y, Nakamura K, Saito M, Narita S, Ogawa O, Habuchi T, Kawamoto H.
  • Journal Title: iScience Volume: 23 Issue: 4 Pages: 100998-100998
  • DOI: 10.1016/j.isci.2020.100998
  • NAID: 120006826710
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Long non-coding RNA MANCR is a target of BET bromodomain protein BRD4 and plays a critical role in cellular migration and invasion abilities of prostate cancer. (2020)
  • Author(s): Nagasawa M, Tomimatsu K, Terada K, Kondo K, Miyazaki K, Miyazaki M, Motooka D, Okuzaki D, Yoshida T, Kageyama S, Kawamoto H, Kawauchi A, Agata Y.
  • Journal Title: Biochem Biophys Res Commun. Volume: 526 Issue: 1 Pages: 128-134
  • DOI: 10.1016/j.bbrc.2020.03.043
  • NAID: 120006839258
  • Peer Reviewed
• [Journal Article] Ca2+ signal is involved in endothelin-1-induced internalization of endothelin type A receptor expressed in Chinese hamster ovary cells (2019)
  • Author(s): Horinouchi Takahiro、Karki Sarita、Terada Koji、Mazaki Yuichi、Miwa Soichi
  • Journal Title: Journal of Pharmacological Sciences Volume: 140 Issue: 1 Pages: 102-105
  • DOI: 10.1016/j.jphs.2019.03.008
  • Peer Reviewed / Open Access
• [Presentation] TCR遺伝子導入iPS細胞からT細胞を再生するための新規の方法：TCRカセット法の開発 (2022)
  • Author(s): 永野誠治、寺田晃士、縣保年、河本宏
  • Organizer: Kyoto T cell Conference 第31回学術集会
• [Presentation] ビタミンCはCD8 T細胞の生存能を亢進させる (2022)
  • Author(s): 近藤健太、長谷川達矢、寺田晃士、口分田美奈、新田信人、前田菜摘、奥崎大介、縣保年
  • Organizer: Kyoto T cell Conference 第31回学術集会
• [Presentation] Development of “TCR cassette method” to transduce pluripotent stem cells with exogenous TCR gene (2022)
  • Author(s): 永野誠治、寺田晃士、縣保年、河本宏
  • Organizer: 第84回日本血液学会学術集会
• [Presentation] Regeneration of antigen specific CTLs from TCR-iPSCs prepared with gene editing: development of “TCR cassette method” (2022)
  • Author(s): 永野誠治、寺田晃士、縣保年、増田喬子、河本宏
  • Organizer: 第26回がん免疫学会総会
• [Presentation] Vitamin C treatment promotes the persistence and immune responses of CD8+ T cells (2022)
  • Author(s): 近藤健太、長谷川達矢、寺田晃士、縣保年
  • Organizer: 第51回日本免疫学会学術集会
• [Presentation] Generation of CTLs from iPSCs transduced with TCR genes: development of “TCR cassette” method (2022)
  • Author(s): 永野誠治、寺田晃士、縣保年、河本宏
  • Organizer: 第51回日本免疫学会学術集会
• [Presentation] Development of adaptive immunotherapy using neoantigen (NeoAg)-specific T cell receptor (TCR)-gene transduced T cells (2022)
  • Author(s): Hidetoshi Sumimoto, Koji Terada, Yasutoshi Agata and Yataro Daigo
  • Organizer: 第19回日本臨床腫瘍学会学術集会
• [Presentation] “TCRカセット法“の開発：がん抗原特異的TCR遺伝子を内在性TCR遺伝子座へ効率よく導入する (2021)
  • Author(s): 寺田晃士，永野誠治，近藤健太，増田喬子，河本宏，縣保年
  • Organizer: Kyoto T cell Conference 第３０回学術集会
• [Presentation] TCR遺伝子導入iPS細胞からCTLを再生するための新規の方法：TCRカセット法の開発 (2021)
  • Author(s): 永野誠治，寺田晃士，縣保年，河本宏
  • Organizer: Kyoto T cell Conference 第３０回学術集会
• [Presentation] カニクイザルの腫瘍浸潤T細胞から腫瘍殺傷能をもつTCR遺伝子を単離する (2021)
  • Author(s): 近藤健太，寺田晃士，石垣宏仁，長嶋彩花，里岡大樹，永野誠治，増田喬子，川村晃久，平田多佳子，小笠原一誠，伊藤靖，河本宏、縣保年
  • Organizer: Kyoto T cell Conference 第３０回学術集会
• [Presentation] Isolation of TCR genes with tumor-killing activity from tumor-infiltrating lymphocytes in a tumor rejection cynomolgus macaque model (2021)
  • Author(s): Koji Terada, Kenta Kondo, Hirohito Ishigaki, Ayaka Nagashima, Hiroki Satooka, Seiji Nagano, Kyoko Masuda, Teruhisa Kawamura, Takako Hirata, Kazumasa Ogasawara, Yasushi Itoh, Hiroshi Kawamoto and Yasutoshi Agata
  • Organizer: 第50回日本免疫学会学術集会
• [Presentation] Development of “TCR cassette method”: Regeneration of CTLs from iPSCs in which tumor-antigen specific TCR genes can be efficiently introduced into the endogenous TCR locus by cassette (2021)
  • Author(s): Koji Terada, Kenta Kondo, Seiji Nagano, Kyoko Masuda, Hiroshi Kawamoto and Yasutoshi Agata
  • Organizer: 第50回日本免疫学会学術集会
• [Presentation] Vitamin C alters gene expression of CD8+ T cells through DNA demethylation (2021)
  • Author(s): Kenta Kondo, Tatsuya Hasegawa, Koji Terada, Yasutoshi Agata
  • Organizer: 第50回日本免疫学会学術集会
• [Presentation] ゲノム編集とカセット交換法を用いたがん抗原特異的TCR遺伝子導入法の開発 (2020)
  • Author(s): 寺田晃士、近藤健太、永野誠治、前田卓也、増田喬子、河本宏、縣保年
  • Organizer: 第２４回がん免疫学会総会
• [Presentation] 近藤健太、寺田晃士、石垣宏仁、里岡大樹、永野誠治、増田喬子、平田多佳子、小笠原一誠、伊藤靖、河本宏、縣保年 (2020)
  • Author(s): カニクイザルの腫瘍浸潤T細胞から腫瘍殺傷能を有するTCR遺伝子を単離する
  • Organizer: 第２４回がん免疫学会総会
• [Presentation] "TCRカセット法"の開発：がん抗原特異的TCR遺伝子を内在性TCR遺伝子座へ効率よく導入できるiPS細胞から細胞傷害性T細胞を再生する (2020)
  • Author(s): 寺田晃士、近藤健太、永野誠治、増田喬子、河本宏、縣保年
  • Organizer: 第43回日本分子生物学会年会
• [Presentation] カニクイザルの腫瘍拒絶モデルにおける腫瘍浸潤T細胞からの腫瘍殺傷能をもつTCR遺伝子の単離 (2020)
  • Author(s): 寺田晃士、石垣宏仁、近藤健太、長嶋彩花、里岡大樹、永野誠治、増田喬子、川村晃久、河本宏、平田多佳子、小笠原一誠、伊藤靖、縣保年
  • Organizer: 第43回日本分子生物学会年会
• [Presentation] Development of an efficient method to introduce TCR genes into the endogenous TCR locus by genome editing and cassette exchange (2019)
  • Author(s): Koji Terada, Ryohei Kondo, Seiji Nagano, Kyoko Masuda, Hiroshi Kawamoto and Yasutoshi Agata
  • Organizer: The 48th Annual Meeting of The Japanese Society for Immunology
• [Presentation] Generation of CTLs from iPSCs transduced with TCR genes: development of “TCR cassette” method (2019)
  • Author(s): Seiji Nagano, Koji Terada, Ryohei Kondo, Yasutoshi Agata, Kyoko Masuda and Hiroshi Kawamoto
  • Organizer: The 48th Annual Meeting of The Japanese Society for Immunology