Development of the novel gene-immunotherapy using artificial CTL targeting leukemia stem cells

• Project/Area Number: 24390245
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Hematology
• Research Institution: Ehime University
• Principal Investigator: YASUKAWA Masaki 愛媛大学, 医学(系)研究科(研究院), 教授 (60127917)
• Co-Investigator(Renkei-kenkyūsha): FUJIWARA Hiroshi 愛媛大学, 医学部附属病院, 講師 (20398291)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥17,810,000 (Direct Cost: ¥13,700,000、Indirect Cost: ¥4,110,000); Fiscal Year 2014: ¥5,720,000 (Direct Cost: ¥4,400,000、Indirect Cost: ¥1,320,000); Fiscal Year 2013: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000); Fiscal Year 2012: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
• Keywords: 白血病 / がん免疫療法 / 細胞傷害性T細胞 / T細胞レセプター / 遺伝子治療 / 細胞障害性Ｔ細胞 / CAR-T細胞 / WT1 / 抗体療法 / 白血病幹細胞 / Ｔ細胞レセプター / ヒト化マウス / CD16 / hTERT / 免疫療法

Outline of Final Research Achievements

We performed the series of experiment in order to establish the novel gene-immunotherapy targeting leukemia stem cells. WT1-TCR-gene-transduced CTLs exerted cytotoxicity against leukemia cells but not normal cells. It was strongly suggested that leukemia stem cells might be eliminated completely by administration of WT1-TCR gene-modified T cells. WT1-specific and HLA-restricted CTLs could be generated in HLA-TG-NSG mice by transplantation of WT1-TCR gene-transduced human hematopoietic stem cells. CD16-CD3ζ chimeric antigen receptor gene-modified CD8+ T cells showed strong ADCC activity in the presence of anti-CD20 monoclonal antibody targeting malignant lymphoma cells. These data strongly suggest that immunotherapy using gene-modified CTLs can establish the cure of chemotherapy-resistant hematological malignancies.

Research Products

• [Journal Article] Antileukemia multifunctionality of CD4+ T cells genetically engineered by HLA class I-restricted and WT1-specific T-cell receptor gene transfer (2015)
  • Author(s): Fujiwara H, Ochi T, Ochi F, Miyazaki Y, Asai H, Narita M, Okamoto S, Mineno J, Kuzushima K, Shiku H, and Yasukawa M
  • Journal Title: Leukemia Volume: in press
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Cytotoxic T lymphocyte lysis of HTLV-1 infected cells is limited by weak HBZ protein expression, but non-specifically enhanced on induction of Tax expression. (2014)
  • Author(s): Rowan AG, Suemori K, Fujiwara H, Yasukawa M, Tanaka Y, Taylor GP, Bangham C.
  • Journal Title: Retrovirology Volume: 11 Pages: 116-116
  • Peer Reviewed / Open Access
• [Journal Article] Gene-Modified Human α/β-T Cells Expressing a Chimeric CD16-CD3ζ Receptor as Adoptively Transferable Effector Cells for Anticancer Monoclonal Antibody Therapy. (2014)
  • Author(s): Yasukawa M, et al.
  • Journal Title: Cancer Immunol Res Volume: 2 Issue: 3 Pages: 249-262
  • DOI: 10.1158/2326-6066.cir-13-0099-t
  • Peer Reviewed / Open Access
• [Journal Article] Abnormal behaviors and developmental disorder of hippocampus in zinc finger protein 521 (ZFP521) mutant mice. (2014)
  • Author(s): Ohkubo N, Matsubara E, Yamanouchi J, Akazawa R, Aoto M, Suzuki Y, Sakai I, Abe T, Kiyonari H, Matsuda S, Yasukawa M, Mitsuda N.
  • Journal Title: PLoS One Volume: 9 Issue: 3 Pages: e92848-e92848
  • DOI: 10.1371/journal.pone.0092848
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Adoptive transfer of genetically engineered WT1-specific cytotoxic T lymphocytes does not induce renal injury (2014)
  • Author(s): Asai H, Fujiwara H, Kitazawa S, Kobayashi N, Ochi T, Miyazaki Y, Ochi F, Akatsuka Y, Okamoto S, Mineno J, Kuzushima K, Ikeda H, Shiku H, Yasukawa M
  • Journal Title: J Hematol Oncol. Volume: 7 Issue: 1 Pages: 3-3
  • DOI: 10.1186/1756-8722-7-3
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Development of a novel redirected T-cell-based adoptive immunotherapy targeting human telomerase reverse transcriptase for adult T-cell leukemia. (2013)
  • Author(s): Yasukawa M, et al.
  • Journal Title: Blood Volume: 121 Issue: 24 Pages: 4894-4901
  • DOI: 10.1182/blood-2012-11-465971
  • Peer Reviewed
• [Journal Article] Adoptive transfer of genetically modified Wilms' tumor 1-specific T cells in a novel malignant skull base meningioma model. (2013)
  • Author(s): Iwami K, Natsume A, Ohno M, Ikeda H, Mineno J, Nukaya I, Okamoto S, Fujiwara H, Yasukawa M, Shiku H, Wakabayashi T.
  • Journal Title: Neuro Oncol. Volume: 15 Issue: 6 Pages: 747-758
  • DOI: 10.1093/neuonc/not007
  • Peer Reviewed
• [Journal Article] Co-introduced functional CCR2 potentiates in vivo anti-lung cancer func tionality mediated by T cells double gene-modified to express WT1-specific T-cell receptor. (2013)
  • Author(s): Yasukawa M, et al.
  • Journal Title: PLoS One Volume: 10 Issue: 2 Pages: e56820-e56820
  • DOI: 10.1371/journal.pone.0056820
  • Peer Reviewed
• [Journal Article] Clinical characteristics and outcomes of Chédiak-Higashi syndrome: a nationwide survey of Japan. (2013)
  • Author(s): Nagai K, Yasukawa M, et al.
  • Journal Title: Pediatr Blood Cancer Volume: 60 Issue: 10 Pages: 1582-1586
  • DOI: 10.1002/pbc.24637
  • Peer Reviewed
• [Journal Article] Protein kinase C inhibitor generates stable human tolerogenic dendritic cells. (2013)
  • Author(s): Matsumoto T, Yasukawa M, et al.
  • Journal Title: J Immunol. Volume: 191 Issue: 5 Pages: 2247-2257
  • DOI: 10.4049/jimmunol.1203053
  • Peer Reviewed
• [Journal Article] Adoptive transfer of genetically modified Wilms' tumor 1-specific T cells in a novel malignant skull base meningioma model. (2013)
  • Author(s): Iwama K, Yasutawa M, et a.
  • Journal Title: Neuro Oncol. Volume: (In press)
  • Peer Reviewed
• [Journal Article] A promising vector for TCR gene therapy: differential effect of siRNA, 2A peptide, and disulfide bond on the introduced TCR expression. (2012)
  • Author(s): Okamoto S, et al.
  • Journal Title: Mol Ther Nucleic Acids. Volume: 18 Pages: e63-e63
  • DOI: 10.1038/mtna.2012.52
  • Peer Reviewed
• [Journal Article] HLA-restricted presentation of WT1 tumor antigen in B-lymphoblastoid cell lines established using a maxi-EBV system. (2012)
  • Author(s): Kanda, T, et al.
  • Journal Title: Cancer Gene Ther. Volume: 19 Issue: 8 Pages: 566-571
  • DOI: 10.1038/cgt.2012.34
  • Peer Reviewed
• [Journal Article] The role of activation-induced cytidine deaminase (AID/AICDA) in the progression of follicular lymphoma (2012)
  • Author(s): Shikata H, Yasukawa M, et al.
  • Journal Title: Cancer Sci. Volume: 103 Pages: 415-421
  • Peer Reviewed
• [Journal Article] Aurora kinase A-specific T-cell receptor gene transfer redirects T-lymphocytes to display effective anti-leukemia reactivity (2012)
  • Author(s): Kozo Nagai, et al
  • Journal Title: Blood Volume: 119(2) Issue: 2 Pages: 368-76
  • DOI: 10.1182/blood-2011-06-360354
  • Peer Reviewed
• [Presentation] Activation Status of Integrin αIIbβ3 in Essential Thrombocythemia with Calreticulin Mutation (2014)
  • Author(s): Jun Yamanouchi , Takaaki Hato, Etsuko Matsubara, Taichi Azuma, Hideyuki Nakanishi, Hiroshi Fujiwara, Yoshihiro Yakushijin, and Masaki Yasukawa
  • Organizer: 56th ASH Annual Meeting
  • Place of Presentation: San Francisco
  • Year and Date: 2014-12-06
• [Presentation] Concomitant Administration of Gene-Modified T Cells Expressing a Chimeric CD16-CD3ζ Receptor with Mogamulizmab Synergistically Suppresses Adult T Cell Leukemia Cells in Vivo (2014)
  • Author(s): Kazushi Tanimoto , Hiroshi Fujiwara, Hiroki Tanaka, Fumihiro Ochi , Hiroaki Asai, Sachiko Okamoto, JunichiMineno , Kiyotaka Kuzushima, Hiroshi Shiku, John Barrett, and Masaki Yasukawa
  • Organizer: 56th ASH Annual Meeting
  • Place of Presentation: San Francisco
  • Year and Date: 2014-12-05
• [Presentation] Redirected CD4+ T Cells towards HLA Class I-Restricted WT1 Epitope Successfully Display Multifactorial Helper Function for the Enhancement of Antileukemia Efficacy Mediated by Redirected T-Cell Based Immunocelltherapy. (2012)
  • Author(s): Miyazaki Y, Yasukawa M, et al.
  • Organizer: 54th Annual Meeting of American Society of Hematology
  • Place of Presentation: Atlanta, USA
  • Year and Date: 2012-12-10
• [Presentation] Human Telomerase Reverse Transcriptase-specific T-Cell Receptor Gene Transfer Redirects T Cells to Display an Effective Antitumor Reactivity Against Adult T-Cell Leukemia (2012)
  • Author(s): Fujiwara H, Yasukawa M, et al.
  • Organizer: 54th Annual Meeting of American Society of Hematology
  • Place of Presentation: Atlanta, USA
  • Year and Date: 2012-12-10
• [Presentation] Therapeutically Infused Redirected T Cells Targeting WT1 Successfully Inhibited Leukemia Stem Cells in Vivo (2012)
  • Author(s): Asai H, Yasukawa M, et al.
  • Organizer: 54th Annual Meeting of American Society of Hematology
  • Place of Presentation: Atlanta, USA
  • Year and Date: 2012-12-10
• [Remarks] 愛媛大学大学院医学系研究科血液・免疫・感染症内科学講座
  • URL: http://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・感染症内科学講座
  • URL: http://www.m.ehime-u.ac.jp/school/int.med1/
• [Remarks]
  • URL: http://www.m.ehime-u.ac.jp/school/int.medl/