Application of iPSCs for hemophilia therapy

2013 Fiscal Year Final Research Report

• Project/Area Number: 23591427
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Hematology
• Research Institution: Jichi Medical University
• Principal Investigator: OHMORI Tsukasa 自治医科大学, 医学部, 講師 (70382843)
• Co-Investigator(Kenkyū-buntansha): MADOIWA Seiji 自治医科大学, 分子病態治療研究センター分子病態研究部, 講師 (70296119)
• Project Period (FY): 2011 – 2013
• Keywords: 遺伝子細胞治療 / 血友病 / 人工多能性幹細胞

Research Abstract

We focused on the use of iPSCs for cell-based therapy of hemophilia. We generated iPSCs from mesenchymal stem cells isolated from C57BL/6 mice. The mouse iPSCs were generated through the induction of four Yamanaka transcription factor genes. The iPSCs released functional coagulation factor VIII (FVIII) following transduction with a lentiviral vector. The subcutaneous transplantation of iPSCs expressing FVIII into nude mice resulted in teratoma formation, and significantly increased plasma levels of FVIII. The plasma concentration of FVIII was at levels appropriate for human therapy at 2–4 weeks post-transplantation. We next examined which promoter was suitable for the production of coagulation factor from iPSCs, and found that EF1alpha promoter stably drove transgene in iPSCs. We also developed iPSCs from FIX-deficient mouse fibroblasts by sendai virus vector. The established iPSCs had a potency to produce chimeric mouse, and could produce functional FIX after the transduction.

Research Products

• [Journal Article] Production of functional coagulation factor VIII from iPSCs using a lentiviral vector (2014)
  • Author(s): Kashiwakura Y, Ohmori T, Mimuro J, Madoiwa S, Inoue M, Hasegawa M, Ozawa K, and Sakata Y
  • Journal Title: Haemophilia Volume: 20 Pages: e40-44
• [Journal Article] Overexpression of factor VII ameliorates bleeding diathesis of factor VIII-deficient mice with inhibitors (2013)
  • Author(s): Yasumoto A, Madoiwa S, Kashiwakura Y, Ishiwata A, Ohmori T, Mizukami H, Ozawa K, Sakata Y, Mimuro J
  • Journal Title: Thromb Res Volume: 131 Pages: 444-449
• [Journal Article] Minimizing the Inhibitory Effect of Neutralizing Antibody for Efficient Gene Expression in the Liver With Adeno-associated Virus 8 Vectors (2013)
  • Author(s): Mimuro J, Mizukami H, Hishikawa S, Ikemoto T, Ishiwata A, Sakata A, Ohmori T, Madoiwa S, Ono F, Ozawa K, Sakata Y
  • Journal Title: Mol Ther Volume: 21 Pages: 318-323
• [Journal Article] Genetically modified adipose tissue-derived stem/stromal cells, using simian immunodeficiency virus-based lentiviral vectors, in the treatment of hemophilia B (2013)
  • Author(s): Watanabe N, Ohashi K, Tatsumi K, Utoh R, Shim IK, Kanegae K, Kashiwakura Y, Ohmori T, Sakata Y, Inoue M, Hasegawa M, Okano T
  • Journal Title: Hum Gene Ther Volume: 24 Pages: 283-294
• [Journal Article] Porcine model of hemophilia A (2012)
  • Author(s): Kashiwakura Y, Mimuro J, Onishi A, Iwamoto M, Madoiwa S, Fuchimoto D, Suzuki S, Suzuki M, Sembon S, Ishiwata A, Yasumoto A, Sakata A, Ohmori T, Hashimoto M, Yazaki S, Sakata Y
  • Journal Title: PLoS One Volume: 7 Pages: e49450
• [Journal Article] Intra-articular injection of mesenchymal stem cells expressing coagulation factor ameliorates hemophilic arthropathy in factor VIII-deficient mice (2012)
  • Author(s): Kashiwakura Y, Ohmori T, Mimuro J, Yasumoto A, Ishiwata A, Sakata A, Madoiwa S, Inoue M, Hasegawa M, Ozawa K, and Sakata Y
  • Journal Title: J Thromb Haemost Volume: 10 Pages: 1802-1813
• [Journal Article] Immune response against serial infusion of factor VIII antigen through an implantable venous-access device system in haemophilia A mice (2012)
  • Author(s): Madoiwa S, Kobayashi E, Kashiwakura Y, Sakata A, Yasumoto A, Ohmori T, Mimuro J, Sakata Y
  • Journal Title: Haemophilia Volume: 18 Pages: e323-330
• [Presentation] Development of a cell-based therapy for treating hemophilic arthropathy by local injection of transduced MSCs (2013)
  • Author(s): 大森司
  • Organizer: 第35回日本血栓止血学会学術集会
  • Place of Presentation: 山形
  • Year and Date: 20130530-0601
• [Presentation] 血友病Aクローンブタ (2013)
  • Author(s): 柏倉裕志、三室淳、大西彰、岩元正樹、窓岩清治、淵本大一郎、鈴木俊一、鈴木美佐枝、千本昭一郎、石渡彰、安本篤史、坂田飛鳥、大森司、橋本径子、矢崎智子、坂田洋一
  • Organizer: 第35回日本血栓止血学会学術集会, (日本血栓止血学会誌)
  • Place of Presentation: 山形
  • Year and Date: 20130530-0601
• [Presentation] 人工多能性幹細胞を用いた新規血友病治療法に対する基礎的検討 (2012)
  • Author(s): 柏倉裕志、大森司、坂田飛鳥、窓岩清治、井上誠、長谷川護、三室淳、坂田洋一
  • Organizer: 第74回日本血液学会学術集会
  • Place of Presentation: 京都
  • Year and Date: 20121019-21
• [Presentation] Dose-respnse relationship of factor IX expression in non-human orimates using AAV8-based vector (2012)
  • Author(s): 水上浩明、三室淳、塚原智典、卜部匡司、久米晃浩、大森司、窓岩清治、坂田洋一、小澤敬也
  • Organizer: 第74回日本血液学会学術集会
  • Place of Presentation: 京都
  • Year and Date: 20121019-21
• [Presentation] FVIII遺伝子導入MSCの関節内移植は血友病Aマウスの血友病性関節症を改善する (2012)
  • Author(s): 柏倉裕志,大森司,三室淳,安本篤史,石渡彰,坂田飛鳥,窓岩清治,井上誠,長谷川護,小澤敬也,坂田洋一
  • Organizer: 第34回日本血栓止血学会学術集会
  • Place of Presentation: 東京
  • Year and Date: 20120607-09
• [Presentation] Intra-articular injection of autologous mesenchymal stem cells ameliorates hemophilic arthropathy in factor VIII-deficient mice (2011)
  • Author(s): Kashiwakura Y, Ohmori T, Mimuro J, Yasumoto A, Ishiwata A, Sakata A, Madoiwa S, Inoue M, Hasegawa M, Watanabe N, Tatsumi K, Ohashi K, Okano T, Sakata Y
  • Organizer: ISTH2011.XXIII Congress of the International Society on Thrombosis and Haemostasis.57^<th> Annual SSC Meeting
  • Year and Date: 20110723-28