gene introduction using myeloid lineage cells

2017 Fiscal Year Annual Research Report

• Project/Area Number: 16K14634
• Research Institution: Teikyo University
• Principal Investigator: 白藤 尚毅 帝京大学, 医学部, 教授 (00206301)
• Project Period (FY): 2016-04-01 – 2018-03-31
• Keywords: CAR-T / PBSCT / G-CSF / CLL-1 / CD19

Outline of Annual Research Achievements

ヒトCD47遺伝子に対するアンチセンスRNA並びにmiRNAを発現するシャトルベクターを構築した。ヒト正常骨髄を採取、単核球層を分離、抗体を用いてCD13, CD33 陽性骨髄芽球分画をmagnet beadで選別し、構築したウイルスベクターを遺伝子導入後human granulocyte colony-stimulating factor を添加後培養し、好中球へ分化、増殖させた（論文準備中）。
この系を用いて細胞の選別や培養に関して条件を変えながら観察を続けた。CD34陽性造血前駆細胞をmagnet beadで選別し、造血幹細胞増殖培地で培養後G-CSFを添加し再び培養を行った後CD13, 33 陽性細胞群を選別、さらG-CSF添加して培養することで最も高い効率が得られた。
以上の観察から現在末梢血幹細胞採取の際患者さんに協力していただき、その1/100量の細胞を用いてin vitroでの治療実験を行っている。ヒト末梢血造血幹・前駆細胞からCD34陽性細胞を分離し、CD19-CAR 及び CLL-1-CAR vectorを感染させ、CD13 及び CD33 陽性細胞を選別する。G-CSFを添加し、CAR-N とする。Positive control としてCD3, 8 陽性 T 細胞を選別、IL-7, IL-15 添加下培養し、活性化した状態でウイルスに感染させCAR-T とする。標的となる細胞は CD19-CAR では Daudi 、CLL-1-CAR では HL60 を標的として現在標準的CAR-Tと各種CAR とで in vitro 治療実験を行っている。研究期間は本春で終了するがかなり優位な結果が得られており本年度も科研費公募を行いながら本研究を行っていく計画である。

Research Products

• [Journal Article] Direct Factor X sequestration by systemic amyloid lightchain amyloidosis (2018)
  • Author(s): Haruko Tashiro, Ryosuke Shirasaki, Masato Watanabe, Kazuo Kawasugi, Yoshihisa Takahashi, Naoki Shirafuji
  • Journal Title: Clinical Case Reports Volume: 6(3): Pages: 513, 515
  • DOI: 10.1002/ccr3.1398
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Aggressive NK Cell Leukemia with Hemophagocytic Lymphohistiocytosis: A Case Report (2017)
  • Author(s): Kimiko Nogi, Haruko Tashiro, Kazuo Kawasugi, Mayumi Matsuzawa, Takeshi Osawa, Ryohei Toyooka, Taiji Furukawa, Naoki Shirafuji
  • Journal Title: J Clin Lab Med Volume: 2.2 Pages: 1, 4
  • DOI: http://dx.doi.org/10.16966/2572-9578.116
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Immunotherapy against cancer-related viruses. (2017)
  • Author(s): Tashiro H, Brenner MK
  • Journal Title: Cell Res. Volume: 27 (1) Pages: 59, 73
  • DOI: 10.1038/cr.2016.153.
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Vaccination targeting native receptors to enhance the function and proliferation of chimeric antigen receptor (CAR)-modified T cells. (2017)
  • Author(s): Tanaka M, Tashiro H, Omer B, Lapteva N, Ando J, Ngo M, Mehta B, Dotti G, Kinchington PR, Leen AM, Rossig C, Rooney CM.
  • Journal Title: Clin Cancer Res. Volume: 23 Pages: 3499-3509.
  • DOI: 10.1158/1078-0432.CCR-16-2138.
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Comprehensive Approach for Identifying the T Cell Subset Origin of CD3 and CD28 Antibody-Activated Chimeric Antigen Receptor-Modified T Cells. J Immunol. 2017 May 26. pii: 1601494. doi: 10.4049/jimmunol.1601494. (2017)
  • Author(s): Schmueck-Henneresse M, Omer B, Shum T, Tashiro H, Mamonkin M, Lapteva N, Sharma S, Rollins L, Dotti G, Reinke P, Volk HD, Rooney CM.
  • Journal Title: J Immunol. pii: 1601494. Volume: 199(1) Pages: 348-362
  • DOI: 10.4049/jimmunol.1601494.
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] CAR T Cells Administered in Combination with Lymphodepletion and PD-1 Inhibition to Patients with Neuroblastoma. (2017)
  • Author(s): Heczey A, Louis CU, Savoldo B, Dakhova O, Durett A, Grilley B, Liu H, Wu MF, Mei Z, Gee A, Mehta B, Zhang H, Mahmood N, Tashiro H, Heslop HE, Dotti G, Rooney CM, Brenner MK.
  • Journal Title: Mol Ther. Volume: 25(9) Pages: 2214-2224
  • DOI: 10.1016/j.ymthe.2017.05.012.
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Treatment of acute myeloid leukemia with T cells expressing chimeric antigen receptors directed to C-type lectin-like molecule 1. (2017)
  • Author(s): Tashiro H, Sauer T, Shum T, Parikh K, Mamonkin M, Omer B, Rouce R, Lulla P, Rooney C, Gottschalk S, Brenner MK. Treatment of acute myeloid leukemia with T cells expressing chimeric antigen receptors directed to C-type lectin-like molecule 1.
  • Journal Title: Mol Ther. Volume: 25 (9) Pages: 2202-2213
  • DOI: https://doi.org/10.1016/j.ymthe.2017.05.024
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] IL-1-βと電気刺激による成人ヒト皮膚線維芽細胞のhemogenic endotheliumへの変化に関する研究 (2018)
  • Author(s): 白藤 尚毅,松尾 琢二,住吉 立,田代 晴子
  • Organizer: 第17回日本再生医療学会総会
• [Presentation] 赤血球抗原情報検索システムの有用性と問題点について (2017)
  • Author(s): 灰野夏澄、前島理恵子、藤原孝記、冨山秀和、金子 強、永友ひとみ、蟹井はるか、笠井英利、難波宏美、大曽根和子、松本謙介、白藤尚毅。
  • Organizer: 第65回日本輸血・細胞治療学会（6月、千葉）
• [Presentation] ABO不適合臍帯血移植後のドナー由来赤血球抗原量の解析 (2017)
  • Author(s): 蟹井はるか、藤原孝記、前島理恵子、冨山秀和、灰野夏澄、金子 強、永友ひとみ、笠井英利、難波宏美、大曽根和子、松本謙介、白藤尚毅。
  • Organizer: 第65回日本輸血・細胞治療学会（6月、千葉）
• [Presentation] 好中球増多を伴った多発性骨髄腫の１例。 (2017)
  • Author(s): 芳賀、幸、松本謙介、住吉 立，松尾 琢二，山本 義，白崎良輔、田代晴子、大井 淳，白藤尚毅、滝川 一。
  • Organizer: 第633回日本内科学会関東地方会 (6月、東京)
• [Presentation] 全身性アミロイドーシスによる高度の第X因子欠乏を来したB細胞性リンパ増殖性疾患 (2017)
  • Author(s): 田代晴子、住吉 立，松尾 琢二，山本 義，松本謙介、大井 淳，秋山 暢，川杉 和夫，白藤尚毅。
  • Organizer: 第7回日本血液学会関東甲信越地方会（7月、松本）
• [Presentation] FLT3-ITD mutation in MDS onset is associated with early transformation to AML (2017)
  • Author(s): Ritsu Sumiyoshi, Tadashi Yamamoto, Takuji Matsuo, Yoko Oka-Miura, Haruko Tashiro, Kensuke Matsumoto, Jun Ooi, Naoki Shirafuji.
  • Organizer: 78回日本血液学会総会（10月、横浜）
  • Int'l Joint Research
• [Presentation] Low HDL is a significant poor prognostic factor in malignant lymphoma (ML) (2017)
  • Author(s): Takuji Matsuo, Ritsu Sumiyoshi, Tadashi Yamamoto, Haruko Tashiro, Kensuke Matsumoto, Jun Ooi, Nobu Akiyama, Kazuo Kawasugi, Naoki Shirafuji.
  • Organizer: 78回日本血液学会総会（10月、横浜）
  • Int'l Joint Research
• [Presentation] Perkinson’s disease (PD) developed after Bortezomib (Bor)-containing chemotherapy for Mantle cell lymphoma (MCL) (2017)
  • Author(s): Yoko Miura, Takuji Matsuo, Ritsu Sumiyoshi, Tadashi Yamamoto, Haruko Tashiro, Kennsuke Matsumoto, Jun Ooi, Naoki Shirafuji.
  • Organizer: 78回日本血液学会総会（10月、横浜）
  • Int'l Joint Research
• [Presentation] Primary Adrenal Lymphoma with Adrenal Insufficiency: Report of Three Cases and Review of Literature (2017)
  • Author(s): Masumi Ogawa, Naoki Edo, Takuji Matsuo, Tadashi Yamamoto, Satoshi Takahashi, Yamato Mashimo, Koji Morita, Yuko Sasajima, Hiroshi Uozaki, Fukuo Kondo, Yasunori Ota, Naoki Shirafuji, Hiroko Okinaga, Kazuhisa Tsukamoto, Toshio Ishikawa.
  • Organizer: ENDO 2017 (April, Orlando, Florida USA)
  • Int'l Joint Research
• [Presentation] Constitutive Signaling from an Engineered IL-7 Receptor Promotes Durable Tumor Elimination by Tumor Redirected T-Cells (2017)
  • Author(s): Thomas Shum, Bilal Omer, Haruko Tashiro, Robert Kruse, Dimitrios Wagner, Kathan Parikh, Zhongzhen Yi, Tim Sauer, Robin Parihar, Leonid Metelitsa, Stephen Gottschalk, Cliona Rooney
  • Organizer: ASGCT 20th Annual Meeting (May, Washington DC, USA)
  • Int'l Joint Research
• [Presentation] A Single CRISPR-Mediated Knockout as Novel Suicide Switch and Selection Tool for Gene-Modified T Cells (2017)
  • Author(s): Dimitrios L. Wagner, Haruko Tashiro, Thomas Shum, Debopriyo Halder, Michael C. Gundry, Lorenzo Brunetti, Ciaran M. Lee, Benjamin H. Shin, Tim Sauer, Maksim Mamonkin, Gang Bao, Margaret A. Goodell, Cliona M. Rooney
  • Organizer: ASGCT 20th Annual Meeting (May, Washington DC, USA)
  • Int'l Joint Research
• [Presentation] Low high-density lipoprotein cholesterol (HDL) is a significant poor prognostic factor in malignant lymphoma (2017)
  • Author(s): Takuji Matsuo, Haruko Tashiro, Ritsu Sumiyoshi, Tadashi Yamamoto, Yoko Oka-Miura, Kensuke Matsumoto, Jun Ooi, Naoki Shirafuji
  • Organizer: 59th ASH meeting (December, Atlanta, Georgia, USA,)
  • Int'l Joint Research
• [Presentation] FLT3-ITD mutation in MDS patients is associated with early transformation to AML (2017)
  • Author(s): Ritsu Sumiyoshi, Haruko Tashiro, Takuji Matsuo, Tadashi Yamamoto, Yoko Oka-Miura, Kensuke Matsumoto, Jun Ooi, Naoki Shirafuji.
  • Organizer: 59th ASH meeting (December, Atlanta, Georgia, USA,)
  • Int'l Joint Research