Enhancement of Intracellular Signaling in T Cell Receptor-transduced T Cells

2020 Fiscal Year Annual Research Report

• Project/Area Number: 18K08351
• Research Institution: Nagoya University
• Principal Investigator: 寺倉 精太郎 名古屋大学, 医学部附属病院, 病院講師 (40625141)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Keywords: TCR遺伝子導入 / T細胞養子免疫療法 / 新規アダプター分子

Outline of Annual Research Achievements

以前に示したadapter分子を用いた遺伝子導入T細胞の機能向上について、１回の遺伝子導入でTCRとadapter分子の双方を遺伝子導入可能とし、実際的な細胞作成法を樹立するため、NY-ESO-1 TCRとadapter分子（ATAM）を１回の遺伝子導入によりT細胞内に遺伝子導入可能とする新規ベクターの開発を試みた。TCR-β鎖およびα鎖とATAMをそれぞれ1:1で発現するall-in-one ベクターを作成した。この方法 (1 virus vector法, 1vv法)ではTCRとATAMが等モルに発現する。一方で先行研究で実績のある二つのウイルスベクターを用いる方法 (2 virus vector法, 2vv法)によるTCR-Tの作成においてはTCRとATAMの発現はATAMの方が遺伝子の長さが短いこともあり、Jurkat細胞に対する二つの方法で作成した1vv-TCR-Tと2vv-TCR-Tを比較したウェスタンブロット法でも、フローサイト メトリー法でも約3倍程度高かった。
1vv法で作成したTCR-T細胞と 2vv法で作成したTCR-T細胞の機能を比較したところ、2vv法で作成したTCR-Tの方が有意に抗原刺激後の増殖が有利であった。このことは刺激後の細胞内シグナルを比較したところ、2vv法で作成したTCR-Tでは有意にリン酸化Erkおよびリン酸化p38のシグナルの増強が見られたことから、裏付けられた。
1vv法による遺伝子導入ではATAMのモル比が、TCRに比べるとほとんど同じになることが示され、この比率ではTCR-Tにおけるシグナル増幅につながらないことがわかった。TCRよりも3倍以上多くのATAMを遺伝子導入する必要があることが示された。

Research Products

• [Int'l Joint Research] Medical University of Vienna(オーストリア)
  • Country Name: AUSTRIA
  • Counterpart Institution: Medical University of Vienna
• [Journal Article] Comparison of immunosuppressant regimens in salvage cord blood transplantation for graft failure after allogeneic hematopoietic stem cell transplantation (2021)
  • Author(s): Harada K, Fuji S, Seo S, Uchida N, Kawakita T, Yano S, Ozawa Y, Yoshioka S, Onishi Y, Noguchi Y, Onizuka M, Matsuhashi Y, Kimura T, Ichinohe T, Atsuta Y, Terakura S, Nakasone H.
  • Journal Title: Bone Marrow Transplant Volume: 56(2) Pages: 400-410
  • DOI: 10.1038/s41409-020-00999-9
  • Peer Reviewed
• [Journal Article] Phase I clinical trial of intra-bone marrow cotransplantation of mesenchymal stem cells in cord blood transplantation (2021)
  • Author(s): Goto T, Murata M, Nishida T, Terakura S, Kamoshita S, Ishikawa Y, Ushijima Y, Adachi Y, Suzuki S, Kato K, Hirakawa A, Nishiwaki S, Nishio N, Takahashi Y, Kodera Y, Matsushita T, Kiyoi H.
  • Journal Title: Stem Cells Transl Med Volume: 10(4) Pages: 542-553
  • DOI: 10.1002/sctm.20-0381
  • Peer Reviewed / Open Access
• [Journal Article] A multicenter phase II study of intrabone single-unit cord blood transplantation without antithymocyte globulin (2021)
  • Author(s): Nishida T, Kobayashi T, Sawa M, Masuda S, Shibasaki Y, Goto T, Fukuhara N, Fujii N, Ikegame K, Sugita J, Ikeda T, Kuwatsuka Y, Suzuki R, Najima Y, Doki N, Kato T, Inagaki Y, Utsu Y, Aotsuka N, Masuko M, Terakura S, Onishi Y, Maeda Y, Okada M, Teshima T, Murata M.
  • Journal Title: Ann Hematol Volume: 100(3) Pages: 743-752
  • DOI: 10.1007/s00277-020-04365-z
  • Peer Reviewed
• [Journal Article] Effect of methotrexate dose in graft-versus-host disease prophylaxis after single-unit cord blood transplantation in adult acute myeloid leukemia (2021)
  • Author(s): Terakura S, Kuwatsuka Y, Sugita J, Takahashi S, Ozawa Y, Ozeki K, Yoshioka S, Nakamae H, Kawakita T, Sawa M, Morishige S, Najima Y, Katsuoka Y, Sakaida E, Kouzai Y, Kimura T, Ichinohe T, Fukuda T, Atsuta Y, Murata M, Teshima T.
  • Journal Title: Int J Hematol Volume: - Pages: -
  • DOI: 10.1007/s12185-021-03097-8
  • Peer Reviewed
• [Journal Article] Single Cord Blood Transplantation Versus Unmanipulated Haploidentical Transplantation for Adults with Acute Myeloid Leukemia in Complete Remission (2021)
  • Author(s): Konuma T, Kanda J, Yamasaki S, Harada K, Shimomura Y, Terakura S, Mizuno S, Uchida N, Tanaka M, Doki N, Ozawa Y, Nakamae H, Sawa M, Matsuoka KI, Morishige S, Maruyama Y, Ikegame K, Kimura T, Kanda Y, Ichinohe T, Atsuta Y, Yanada M.
  • Journal Title: Transplant Cell Ther Volume: 27(4) Pages: 334.e1-334
  • DOI: 10.1016/j.jtct.2021.01.023
  • Peer Reviewed
• [Journal Article] Comparison of Transplantation Outcomes after Foscarnet and Ganciclovir Administration as First-Line Anti-Cytomegalovirus Preemptive Therapy (2021)
  • Author(s): Miyao K, Terakura S, Ozawa Y, Sawa M, Kohno A, Kasahara S, Iida H, Ino K, Kusumoto S, Kasai M, Takami A, Kurahashi S, Kajiguchi T, Morishita T, Nishida T, Murata M; Nagoya Blood and Marrow Transplantation Group.
  • Journal Title: Transplant Cell Ther Volume: 27(4) Pages: 342.e1-342
  • DOI: 10.1016/j.jtct.2020.12.012
  • Peer Reviewed
• [Journal Article] Prospective evaluation of alternative donor from unrelated donor and cord blood in adult acute leukemia and myelodysplastic syndrome (2020)
  • Author(s): Terakura S, Nishida T, Sawa M, Kato T, Miyao K, Ozawa Y, Goto T, Kohno A, Ozeki K, Onishi Y, Fukuhara N, Fujii N, Yokoyama H, Kasai M, Iida H, Kanemura N, Endo T, Ago H, Onizuka M, Iyama S, Nawa Y, Nakamae M, Nagata Y, Kurahashi S, Tomiya Y, Yanagisawa A, Suzuki R, Kuwatsuka Y, Atsuta Y, Miyamura K, Murata M
  • Journal Title: Bone Marrow Transplant Volume: 55(7) Pages: 1399-1409
  • DOI: 10.1038/s41409-020-0859-8
  • Peer Reviewed
• [Journal Article] Comparison of reduced-intensity/toxicity conditioning regimens for umbilical cord blood transplantation for lymphoid malignancies (2020)
  • Author(s): Imahashi N, Terakura S, Kondo E, Kako S, Uchida N, Kobayashi H, Inamoto Y, Sakai H, Tanaka M, Ishikawa J, Kozai Y, Matsuoka KI, Kimura T, Fukuda T, Atsuta Y, Kanda J; Donor/Source Working Group of the Japan Society for Hematopoietic Cell Transplantation.
  • Journal Title: Bone Marrow Transplant Volume: 55(11) Pages: 2098-2108
  • DOI: 10.1038/s41409-020-0905-6
  • Peer Reviewed
• [Journal Article] Staurosporine and venetoclax induce the caspase-dependent proteolysis of MEF2D-fusion proteins and apoptosis in MEF2D-fusion (+) ALL cells (2020)
  • Author(s): Tange N, Hayakawa F, Yasuda T, Odaira K, Yamamoto H, Hirano D, Sakai T, Terakura S, Tsuzuki S, Kiyoi H.
  • Journal Title: Biomed Pharmacother Volume: 128 Pages: 110330
  • DOI: 10.1016/j.biopha.2020.110330
  • Peer Reviewed
• [Journal Article] Safety and efficacy of anti-programmed cell death-1 monoclonal antibodies before and after allogeneic hematopoietic cell transplantation for relapsed or refractory Hodgkin lymphoma: a multicenter retrospective study (2020)
  • Author(s): Ito A, Kim SW, Matsuoka KI, Kawakita T, Tanaka T, Terakura S, Iida H, Nakamae M, Higuchi K, Tamura S, Yoshioka S, Togitani K, Kawano N, Suzuki R, Suzumiya J, Izutsu K, Teshima T, Fukuda T et al.
  • Journal Title: Int J Hematol Volume: 112(5) Pages: 674-689
  • DOI: 10.1007/s12185-020-02960-4
  • Peer Reviewed
• [Journal Article] Updated Comparison of 7/8 HLA Allele-Matched Unrelated Bone Marrow Transplantation and Single-Unit Umbilical Cord Blood Transplantation as Alternative Donors in Adults with Acute Leukemia (2020)
  • Author(s): Miyao K, Terakura S, Kimura F, Konuma T, Miyamura K, Yanada M, Kako S, Morhishima S, Uchida N, Toya T, Ozawa Y, Fukuda T, Tanaka M, Sawa M, Takada S, Yoshida S, Kimura T, Ichinohe T, Atsuta Y, Kanda J; Donor/Source Working Group of the Japan Society for Hematopoietic Cell Transplantation.
  • Journal Title: Biol Blood Marrow Transplant Volume: 26(11) Pages: 2105-2114
  • DOI: 10.1016/j.bbmt.2020.08.001
  • Peer Reviewed
• [Journal Article] Artificial T Cell Adaptor Molecule-Transduced TCR-T Cells Demonstrated Improved Proliferation Only When Transduced in a Higher Intensity (2020)
  • Author(s): Sakai T, Terakura S, Miyao K, Okuno S, Adachi Y, Umemura K, Julamanee J, Watanabe K, Hamana H, Kishi H, Leitner J, Steinberger P, Nishida T, Murata M, Kiyoi H.
  • Journal Title: Mol Ther Oncolytics Volume: 18 Pages: 613-622
  • DOI: 10.1016/j.omto.2020.08.014
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Randomised controlled trial of conditioning regimen for cord blood transplantation for adult myeloid malignancies comparing high-dose cytarabine/cyclophosphamide/total body irradiation with versus without G-CSF priming: G-CONCORD study protocol (2020)
  • Author(s): Terakura S, Konuma T, Tanaka M, Ozawa Y, Onizuka M, Nanno S, Onishi Y, Aotsuka N, Kondo T, Kawakita T, Kato J, Kobayashi T, Nishida T, Yamaguchi T, Kuwatsuka Y, Takahashi S.
  • Journal Title: BMJ Open Volume: 10(12) Pages: e040467
  • DOI: 10.1136/bmjopen-2020-040467
  • Peer Reviewed / Open Access
• [Journal Article] Effect of cytomegalovirus reactivation with or without acute graft-versus-host disease on the risk of nonrelapse mortality (2020)
  • Author(s): Akahoshi Y, Kimura SI, Inamoto Y, Seo S, Muranushi H, Shimizu H, Ozawa Y, Tanaka M, Uchida N, Kanda Y, Katayama Y, Shiratori S, Ota S, Matsuoka KI, Onizuka M, Fukuda T, Atsuta Y, Murata M, Terakura S, Nakasone H.
  • Journal Title: Clin Infect Dis Volume: - Pages: ciaa1871
  • DOI: 10.1093/cid/ciaa1871
  • Peer Reviewed
• [Presentation] 遺伝子改変CD3z人工アダプター分子のTCRに対する至適遺伝子導入比率の検討. (2020)
  • Author(s): 堺 寿保, 寺倉精太郎, 奥野真吾, 宮尾康太郎, 浜名 洋, 岸 裕幸, 西田徹也, 村田 誠, 清井 仁.
  • Organizer: 第12回日本血液疾患免疫療法学会学術集会
• [Presentation] 遺伝子改変CD3z人工アダプター分子を用いた新規機能増幅型TCRベクターの開発 (2020)
  • Author(s): 堺寿保、寺倉精太郎、奥野真吾、宮尾康太郎、浜名洋、岸裕幸、西田徹也、村田誠、清井仁
  • Organizer: 第82回日本血液学会学術集会
• [Presentation] CD37CAR-T細胞の開発における抗原認識の最適化 (2020)
  • Author(s): 奥野真吾、安達慶高、寺倉精太郎、Jakrawadee Julamanee, 堺寿保、西田徹也、村田誠、清井仁
  • Organizer: 第82回日本血液学会学術集会
• [Presentation] 成人急性骨髄性白血病に対する臍帯血移植後のGVHD予防におけるメソトレキサート投与量の影響. (2020)
  • Author(s): 寺倉 精太郎、鍬塚 八千代、杉田 純一、高橋 聡、小澤 幸泰、尾関 和貴、吉岡 聡、中前 博久、河北 敏郎、澤 正史、森重 聡、名島 悠峰、勝岡 優奈、堺田 惠美子、香西 康司、木村 貴文、一戸 辰夫、福田 隆浩、熱田 由子、村田 誠、豊嶋 崇徳.
  • Organizer: 第43回日本造血細胞移植学会総会
• [Patent(Industrial Property Rights)] キメラ抗原受容体 (2020)
  • Inventor(s): Seitaro Terakura
  • Industrial Property Rights Holder: 名古屋大学
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: C20200255JP#P01
  • Overseas