Development and clinical application of cancer-specific single-chain antibody using new technology of antibody modification

• Project/Area Number: 18H02839
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 54010:Hematology and medical oncology-related
• Research Institution: Ehime Prefectural University of Health Science (2020); Ehime University (2018-2019)
• Principal Investigator: YASUKAWA MASAKI 愛媛県立医療技術大学, 保健科学部, 教授 (60127917)
• Co-Investigator(Kenkyū-buntansha): 越智 俊元 愛媛大学, 医学系研究科, 講師 (10571086)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥17,550,000 (Direct Cost: ¥13,500,000、Indirect Cost: ¥4,050,000); Fiscal Year 2020: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2019: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000); Fiscal Year 2018: ¥7,280,000 (Direct Cost: ¥5,600,000、Indirect Cost: ¥1,680,000)
• Keywords: がん免疫療法 / CAR-T細胞 / 一本鎖抗体 / キメラ抗原受容体 (CAR) / 一本鎖抗体ライブラリー / NY-ESO-1 / CD19 / 遺伝子改変T細胞 / 改変抗体 / 二重特異性抗体 / 抗体改変技術 / 遺伝子改変 / キメラ抗原受容体

Outline of Final Research Achievements

In this study, we have generated a single-chain antibody generation system in which CAR-library T cells have been screened based on their antitumor functions. A variable region library obtained from the human immunoglobulin was fused with that of an existing antitumor antibody to generate an scFv library. CAR-library T cells which express the scFv library were stimulated with tumor cells to enrich antigen-specific population. As the results, target-specific recognition of newly generated scFv-expressing CAR-T cells could be finely tuned by changing a new variable region. Moreover, scFv-optimized CAR-T cells showed sufficient antitumor cytotoxicity and better proliferation capacity with long-lived T-cell phenotype, resulting in enhanced antitumor effects in humanized mouse in vivo system. Collectively, this system can be applied to generate a new scFv optimal for each CAR-T cell targeting a variety of surface tumor antigens, resulting in further advancement of CAR-T-cell therapy.

Academic Significance and Societal Importance of the Research Achievements

今回我々が確立した新規一本鎖抗体 (scFv)作製技術を用いれば、CAR-T細胞が発現するscFvの標的認識様式を繊細に調律しながら、CAR-T細胞の機能と抗腫瘍効果を最大限に誘導できるscFvを迅速かつ安定して作製することが可能となる。本研究は、これまでと異なる視点から、CAR-T細胞が標的抗原を認識するために重要な働きを担うscFvに着目した。本技術は、これまでに開発されてきた様々な改良型CAR-T細胞の機能をさらに高めることにも応用が可能であることから、難治性がんを標的とした次世代CAR-T細胞療法開発領域におけるインパクトは特に大きく、社会的意義も高いと考えられる。

Research Products

• [Journal Article] A single-chain antibody generation system yielding CAR-T cells with superior antitumor function (2021)
  • Author(s): Ochi Toshiki、Maruta Masaki、Tanimoto Kazushi、Kondo Fumitake、Yamamoto Toshihiro、Kurata Mie、Fujiwara Hiroshi、Masumoto Junya、Takenaka Katsuto、Yasukawa Masaki
  • Journal Title: Communications Biology Volume: 4 Issue: 1 Pages: 273-273
  • DOI: 10.1038/s42003-021-01791-1
  • Peer Reviewed / Open Access
• [Journal Article] A multicenter non-randomized, uncontrolled single arm trial for evaluation of the efficacy and the safety of the treatment with favipiravir for patients with severe fever with thrombocytopenia syndrome (2021)
  • Author(s): Suemori K, Saijo M, Yamanaka A, Himeji D, Kawamura M, Haku T, Hidaka M, Kamikokuryo C, Kakihana Y, Azuma T, Takenaka K, Takahashi T, Furumoto A, Ishimaru T, Ishida M, Kaneko M, Kadowaki N, Ikeda K, Sakabe S, Taniguchi T, Ohge H, Kurosu T, Yoshikawa T, Shimojima M, Yasukawa M.
  • Journal Title: PLoS Negl Trop Dis. Volume: 15 Issue: 2 Pages: e0009103-e0009103
  • DOI: 10.1371/journal.pntd.0009103
  • Peer Reviewed / Open Access
• [Journal Article] A clinically applicable and scalable method to regenerate T-cells from iPSCs for off-the-shelf T-cell immunotherapy (2021)
  • Author(s): Iriguchi Shoichi、Yasui Yutaka、Kawai Yohei、Arima Suguru、Kunitomo Mihoko、Sato Takayuki、Ueda Tatsuki、Minagawa Atsutaka、Mishima Yuta、Yanagawa Nariaki、Baba Yuji、Miyake Yasuyuki、Nakayama Kazuhide、Takiguchi Maiko、Shinohara Tokuyuki、Nakatsura Tetsuya、Yasukawa Masaki、Kassai Yoshiaki、Hayashi Akira、Kaneko Shin
  • Journal Title: Nature Communications Volume: 12 Issue: 1 Pages: 430-430
  • DOI: 10.1038/s41467-020-20658-3
  • NAID: 120006954089
  • 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] グルタミン代謝によるCD8 T 細胞抗腫瘍活性の調節 (2020)
  • Author(s): 鈴木淳平、名部彰悟、安川正貴、山下政克
  • Journal Title: 癌と化学療法 Volume: 47 Pages: 11-15
• [Journal Article] Direct comparisoin of target-reactivity and cross-reactivity induced by CAR-and BiTE-redirected T cells for the development of antibody-based T-cell therapy. (2019)
  • Author(s): Masaki Maruta, Toshiki Ochi, Kazushi Tanimoto, Hiroaki Asai, et al.
  • Journal Title: Scientific reports Volume: 9 Issue: 1 Pages: 13293-13293
  • DOI: 10.1038/s41598-019-49834-2
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Histone H3K27 Demethylase Negatively Controls the Memory Formation of Antigen-Stimulated CD8+ T Cells (2019)
  • Author(s): Yamada Takeshi、Nabe Shogo、Toriyama Koji、Suzuki Junpei、Inoue Kazuki、Imai Yuuki、Shiraishi Atsushi、Takenaka Katsuto、Yasukawa Masaki、Yamashita Masakatsu
  • Journal Title: The Journal of Immunology Volume: 202 Issue: 4 Pages: 1088-1098
  • DOI: 10.4049/jimmunol.1801083
  • Peer Reviewed / Open Access
• [Journal Article] 造血器腫瘍に対する免疫療法 新たな標的抗原とその治療応用 (2019)
  • Author(s): 越智俊元, 安川正貴
  • Journal Title: 実験医学 Volume: 37 Pages: 2622-2626
• [Journal Article] Safe childbirth for a type 1 antithrombin-deficient woman with novel mutation in the SERPINC1 gene undergoing antithrombin concentrate therapy. (2019)
  • Author(s): Ikeda Y, Yamanouchi J, Hato T, Yasukawa M, Takenaka K
  • Journal Title: Blood Coagul Fibrinolysis Volume: 30 Issue: 1 Pages: 47-51
  • DOI: 10.1097/mbc.0000000000000785
  • Peer Reviewed
• [Journal Article] Development of an FVIII Inhibitor in a Mild Hemophilia Patient with a Phe595Cys Mutation (2018)
  • Author(s): Yamanouchi J, Tokumoto D, Ikeda Y, Maruta M, Kaneko M, Hato T, and Yasukawa M.
  • Journal Title: Internal Medicine Volume: 57 Issue: 21 Pages: 3179-3182
  • DOI: 10.2169/internalmedicine.1145-18
  • ISSN: 0918-2918, 1349-7235
  • Year and Date: 2018-11-01
  • Peer Reviewed
• [Journal Article] Enhancing T cell Receptor Stability in Rejuvenated iPSC-derived T cells improves their use in cancer immunotherapy (2018)
  • Author(s): Minagawa A, Yoshikawa T, Yasukawa M, Hotta A, Kunitomo M, Iriguchi S, Takiguchi M, Kassai Y, Imai E, Yasui Y, Kawai Y, Zhang R, Uemura Y, Miyoshi H, Nakanishi M, Watanabe A, Hayashi A, Kawana K, Fujii T, Nakatsura T, Kaneko S
  • Journal Title: Cell Stem Cell Volume: 23 Issue: 6 Pages: 850-858
  • DOI: 10.1016/j.stem.2018.10.005
  • Peer Reviewed / Open Access
• [Journal Article] Association of platelet response to cilostazol with clinical outcome and CYP genotype in patients with cerebral infarction. (2018)
  • Author(s): Ikeda Y, Yamanouchi J, Kumon Y, Yasukawa M, Hato T
  • Journal Title: Thromb Res Volume: 172 Pages: 14-20
  • DOI: 10.1016/j.thromres.2018.10.003
  • Peer Reviewed
• [Journal Article] Reinforce the antitumor activity of CD8+ T cells via glutamine restriction (2018)
  • Author(s): Nabe Shogo、Yamada Takeshi、Suzuki Junpei、Toriyama Koji、Yasuoka Toshiaki、Kuwahara Makoto、Shiraishi Atsushi、Takenaka Katsuto、Yasukawa Masaki、Yamashita Masakatsu
  • Journal Title: Cancer Science Volume: 109 Issue: 12 Pages: 3737-3750
  • DOI: 10.1111/cas.13827
  • Peer Reviewed / Open Access
• [Journal Article] The tumor suppressor menin prevents effector CD8 T-cell dysfunction by targeting mTORC1-dependent metabolic activation (2018)
  • Author(s): Suzuki Junpei、Yamada Takeshi、Inoue Kazuki、Nabe Shogo、Kuwahara Makoto、Takemori Nobuaki、Takemori Ayako、Matsuda Seiji、Kanoh Makoto、Imai Yuuki、Yasukawa Masaki、Yamashita Masakatsu
  • Journal Title: Nature Communications Volume: 9 Issue: 1 Pages: 1-12
  • DOI: 10.1038/s41467-018-05854-6
  • Peer Reviewed / Open Access
• [Presentation] Fine-tuned scFv-expressing CAR-T cells show marked antitumor reactivity in vivo. (2020)
  • Author(s): Ochi T, Maruta M, Tanimoto K, Kurata M, Masumoto J, Takenaka K, Yasukawa M.
  • Organizer: The 82nd JSH Annual Meeting
• [Presentation] Optimized scFv-expressing CAR-T cells showed marked antitumor reactivity in vivo (2020)
  • Author(s): Ochi T, Maruta M, Tanimoto K, Takenaka K, Yasukawa M.
  • Organizer: 第24回日本がん免疫学会学術集会
• [Presentation] 次世代型一本鎖抗体作製技術に基づき調整したCAR-T細胞を用いた有効かつ安全な免疫療法の開発 (2020)
  • Author(s): 越智俊元, 丸田雅樹, 谷本一史, 倉田美恵, 増本純也, 竹中克斗, 安川正貴.
  • Organizer: 第12回日本血液疾患免疫療法学会学術集会
• [Presentation] Establishment of a novel T-cell-based scFv screening technology to advance modified antibody-based immunotherapy (2019)
  • Author(s): Ochi, T., Maruta, M., Tanimoto, K., Fujiwara, H., Takenaka, K., Yasukawa, M.
  • Organizer: The 17 th Annual Meeting of the Association for Cancer Immunotherapy (CIMT)
  • Int'l Joint Research
• [Presentation] Next-generation CAR T-cell therapy using antitumor scFvs optimized by a cell-based screening system (2019)
  • Author(s): Ochi, T., Maruta, M., Tanimoto, K., Fujiwara, H., Takenaka, K., Yasukawa, M.
  • Organizer: The 81 st Annual Meeting of the Japanese Society of Hematology
  • Invited
• [Presentation] Development of anti-myeloma immunotherapy by exploiting modified antibodies specific for A2/NY-ESO-1 (2018)
  • Author(s): 4)Ochi, T., Maruta, M., Tanimoto, K., Asai, H., Saitou, T., Yakushijin, Y., Fujiwara, H., Imamura,T., Takenaka, K., Yasukawa, M.
  • Organizer: The fourth CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference
  • Int'l Joint Research
• [Presentation] A Heterozygous Mutation of G-Protein-Coupled Receptor 25 in a Family with Inherited Thrombocytopenia and Thrombosis (2018)
  • Author(s): 5)Yamanouchi, J., Ikeda, Y., Hato, T., Yasukawa, M., Takenaka, K.
  • Organizer: 60th ASH Annual Meeting
  • Int'l Joint Research
• [Presentation] Clinical Study on Favipiravir in Patients with Severe Fever with Thrombocytopenia Syndrome (2018)
  • Author(s): Yasukawa, M.
  • Organizer: Nikkei Asian Conference on Communicable Diseases 2018
  • Int'l Joint Research / Invited
• [Presentation] グルタミン代謝抑制によるCD8+ T細胞の抗腫瘍活性増強メカニズム (2018)
  • Author(s): 名部彰悟, 山田武司, 鈴木淳平, 山下政克, 安川正貴
  • Organizer: 第22回がん免疫学会総会
• [Presentation] 改変抗体を用いた難治性骨髄腫に対する新規T細胞免疫療法の開発 (2018)
  • Author(s): 丸田雅樹, 越智俊元, 谷本一史, 東太地, 齋藤卓, 藤原弘, 今村健志, 安川正貴
  • Organizer: 第22回日本がん免疫学会総会
• [Presentation] 改変抗体を応用した難治性骨髄腫に対する新規T細胞免疫療法の開発 (2018)
  • Author(s): 越智俊元, 丸田雅樹, 谷本一史, 朝井洋晶, 斉藤卓, 藤原弘, 今村健志, 竹中克斗, 安川正貴
  • Organizer: 第10回血液疾患免疫療法学会学術集会
• [Presentation] 難治性骨髄腫を標的とした改変抗体T細胞免疫療法 (2018)
  • Author(s): 丸田雅樹, 越智俊元, 谷本一史, 東太地, 斉藤卓, 藤原弘, 今村健志, 安川正貴
  • Organizer: 第80回日本血液学会学術集会
• [Book] 新臨床内科学第10版「HTLV-1感染症」 (2020)
  • Author(s): 安川正貴
  • Total Pages: 3
  • Publisher: 医学書院
• [Remarks] 愛媛大学大学院医学系研究科血液・免疫・感染症内科学ホームページ
  • URL: https://www.m.ehime-u.ac.jp/school/int.med1/
• [Remarks] 愛媛大学大学院医学系研究科血液・免疫・感染症内科学
  • URL: https://www.m.ehime-u.ac.jp/school/int.med1/
• [Patent(Industrial Property Rights)] CARライブラリおよびscFvの製造方法 (2019)
  • Inventor(s): 越智俊元、安川正貴、竹中克斗、藤原弘
  • Industrial Property Rights Holder: 越智俊元、安川正貴、竹中克斗、藤原弘
  • Industrial Property Rights Type: 特許
  • Filing Date: 2019
• [Patent(Industrial Property Rights)] CARライブラリ、scFvのスクリーニング方法、抗体またはその結合断片、キメラ抗原受容体、核酸、細胞、および細胞の製造方法 (2019)
  • Inventor(s): 越智俊元、安川正貴、藤原 弘
  • Industrial Property Rights Holder: 越智俊元、安川正貴、藤原 弘
  • Industrial Property Rights Type: 特許
  • Filing Date: 2019