Application of iPSCs for hemophilia therapy

• 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) ; 三室 淳 自治医科大学, 医学部, 准教授 (10221607)
• Project Period (FY): 2011 – 2013
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2013: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2012: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2011: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: 遺伝子細胞治療 / 血友病 / 人工多能性幹細胞 / iPS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
• [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 Issue: 1
  • DOI: 10.1111/hae.12311
  • Peer Reviewed
• [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
• [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
• [Journal Article] Intra-articular injection of mesenchymal stem cells expressing coagulation factorameliorates hemophilic arthropathy in factor VIII-deficient mice. (2012)
  • Author(s): Kashiwakura, Y
  • Journal Title: J Thromb Haemost. Volume: 10(9) Issue: 9 Pages: 1802-1813
  • DOI: 10.1111/j.1538-7836.2012.04851.x
  • Peer Reviewed
• [Journal Article] Porcine model of hemophilia a. PLoS One. (2012)
  • Author(s): Kashiwakura Y
  • Journal Title: PLoS One. Volume: 7(11) Issue: 11 Pages: e49450-e49450
  • DOI: 10.1371/journal.pone.0049450
  • Peer Reviewed
• [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., et al.
  • Journal Title: Haemophilia Volume: 18 Issue: 3
  • DOI: 10.1111/j.1365-2516.2011.02686.x
  • Peer Reviewed
• [Journal Article] Lack of association between serum paraoxonase-1 activity and residual platelet aggregation during dual anti-platelet therapy (2012)
  • Author(s): Ohmori, T., et al.
  • Journal Title: Thromb Res Volume: 129 Issue: 4 Pages: e36-e40
  • DOI: 10.1016/j.thromres.2011.10.033
  • Peer Reviewed
• [Journal Article] FTY720 improves functional recovery after spinal cord injury by primarily nonimmunomodulatory mechanisms (2012)
  • Author(s): Norimatsu Y, Ohmori T, Kimura A, Madoiwa S, Mimuro J, Seichi A, Yatomi Y, Hoshino Y, Sakata Y.
  • Journal Title: Am J Pathol Volume: 180(4) Issue: 4 Pages: 1625-35
  • DOI: 10.1016/j.ajpath.2011.12.012
  • Peer Reviewed
• [Journal Article] Degradation of cross-linked fibrin by leukocyte elastase as alternative pathway for plasmin-mediated fibrinolysis in sepsis-induced disseminated intravascular coagulation. (2011)
  • Author(s): Madoiwa, S. Tanaka, H. Nagahama, Y. Dokai, M. Kashiwakura, Y. Ishiwata, A. Sakata, A. Yasumoto, A. Ohmori, T. Mimuro, J. Sakata, Y.
  • Journal Title: Thromb Res Volume: 127 Issue: 4 Pages: 349-355
  • DOI: 10.1016/j.thromres.2010.12.008
  • Peer Reviewed
• [Journal Article] Local regulation of neutrophil elastase activity by endogenous alpha1-antitrypsin in lipopolysaccharide-primed hematological cells. (2011)
  • Author(s): Dokai, M., Madoiwa, S., Yasumoto, A., Kashiwakura, Y., Ishiwata, A., Sakata, A., Makino, N., Ohmori, T., Mimuro, J., and Sakata, Y.
  • Journal Title: Thromb Res Volume: 128 Issue: 3 Pages: 283-292
  • DOI: 10.1016/j.thromres.2011.04.024
  • Peer Reviewed
• [Journal Article] Predictive blood coagulation markers for early diagnosis of venous thromboembolism after total knee joint replacement (2011)
  • Author(s): Watanabe, H. Madoiwa, S. Sekiya, H. Nagahama, Y. Matsuura, S. Kariya, Y. Ohmori, T. Mimuro, J. Hoshino, Y. Hayasaka, S. Sakata, Y.
  • Journal Title: Thromb Res Volume: 128 Issue: 6 Pages: e137-e143
  • DOI: 10.1016/j.thromres.2011.07.030
  • Peer Reviewed
• [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
• [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
• [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
• [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
• [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
• [Presentation] 第VIII因子感作血友病マウスにおけるマイクロポートを用いた連続的抗原投与モデルの構築 (2012)
  • Author(s): 窓岩 清治，小林 英司，坂田 飛鳥，柏倉 裕志，大森 司，三室 淳，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
• [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
• [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 and Sakata Y
  • Organizer: XXIII Congress of the International Society on Thrombosis and Haemostasis
  • Place of Presentation: Kyoto
• [Presentation] Application of lentiviral vectors to therapies for inherited bleeding disorders and research into blood cell signal transduction (2011)
  • Author(s): Ohmori T
  • Organizer: Aso International Meeting(招待講演)
  • Place of Presentation: Kumamoto
• [Presentation] Overexpression of factor VII ameliorates bleeding diathesis of FVIII-deficient mice with inhibitors
  • Author(s): 安本 篤史、窓岩 清治、柏倉 裕志、石渡 彰、大森 司、水上 浩明 、小澤 敬也、坂田 洋一 、三室 淳
  • Organizer: 第75回日本血液学会学術集会
  • Place of Presentation: 札幌
• [Presentation] Development of cell-based therapy for treating hemophilic arthropathy by local injection of transduced MSCs
  • Author(s): Ohmori T, Kashiwakura Y, Mimuro J, Yasumoto A, Ishiwata A, Sakata A, Madoiwa S, Inoue M, Hasegawa M, Ozawa K, and Sakata Y
  • Organizer: 第35回日本血栓止血学会学術集会
  • Place of Presentation: 山形
  • Invited
• [Presentation] 血友病Aクローンブタ
  • Author(s): 柏倉裕志，三室 淳，大西 彰，岩本正樹，窓岩清治，渕本大一郎，鈴木俊一，鈴木美佐枝，千本昭一郎，石渡 彰，安本 篤，坂田飛鳥，大森 司，橋本経子，矢崎智子，坂田洋一
  • Organizer: 第35回日本血栓止血学会学術集会
  • Place of Presentation: 山形
• [Book] Stem Cells and Cancer Stem Cells, volume 2 - Chapter 11 (2011)
  • Author(s): Ohmori T, and Sakata Y
  • Total Pages: 8
  • Publisher: Springer