Development of gene therapy approaches towards hemophilia A

• Project/Area Number: 25461460
• 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: Mizukami Hiroaki 自治医科大学, 医学部, 教授 (20311938)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2014: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2013: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: AAVベクター / 血友病 / 遺伝子治療 / ベクター / AAV

Outline of Final Research Achievements

We tested the efficacy of factor VIII expression using AAV vectors to achieve gene therapy for hemophilia A. As the length of factor VIII gene exceeds the packaging capacity of AAV, we constructed vectors using essential parts of factor VIII gene, combined with short, liver-specific promoter. Expression of factor VIII were achieved both in vitro and in vivo (mouse) experiments. On the other hand, the efficiency of vector production was much lower than the regular sized vectors. Further investigation is required to optimize the vector structure, mainly testing shorter promoter elements to improve the vector production rate.

Research Products

• [Journal Article] Long-term two-photon calcium imaging of neuronal populations with subcellular resolution in adult non-human primates (2015)
  • Author(s): Sadakane, O., Masamizu, Y., Watakabe, A., Terada, S., Ohtsuka, M., Takaji, M., Mizukami, H., Ozawa, K., Kawasaki, H., Matsuzaki, M., Yamamori, T.
  • Journal Title: Cell Reports Volume: 13 Pages: 1989-99
  • NAID: 120005829368
  • Peer Reviewed
• [Journal Article] The angiogenesis regulator vasohibin-1 inhibits ovarian cancer growth and peritoneal dissemination and prolongs host survival. (2015)
  • Author(s): Takahashi, Y., Saga, Y., Koyanagi, T., Takei, Y., Machida, S., Taneichi, A., Mizukami, H., Sato, Y., Matsubara, S., Fujiwara, H.
  • Journal Title: Int J Oncol Volume: 47 Pages: 2057-63
  • Peer Reviewed
• [Journal Article] The Tol2 transposon system mediates the genetic engineering of T-cells with CD19-specific chimeric antigen receptors for B-cell malignancies. (2015)
  • Author(s): Tsukahara, T., Iwase, N., Kawakami, K., Iwasaki, M., Yamamoto, C., Ohmine, K.,Uchibori, R., Teruya, T., Ido, H., Yasushi, S., Urabe, M., Mizukami, H., Kume, A., Nakamura, M., Brentjens, R., Ozawa, K.
  • Journal Title: Gene Ther Volume: 22 Pages: 209-15
  • Peer Reviewed
• [Journal Article] Comparative analyses of adeno-associated viral vector serotypes 1, 2, 5, 8 and 9 in marmoset, mouse and macaque cerebral cortex. (2015)
  • Author(s): Watakabe, A., Ohtsuka, M., Kinoshita, M., Takaji, M., Isa, K., Mizukami, H., Ozawa, K., Isa, T., Yamamori, T.
  • Journal Title: Neurosci Res Volume: 93 Pages: 144-57
  • Peer Reviewed
• [Journal Article] The prevalence of neutralizing antibodies against adeno-associated virus capsids is reduced in young Japanese individuals. (2014)
  • Author(s): Mimuro J, Mizukami H, Shima M, Matsushita T, Taki M, Muto S, Higasa S, Sakai M, Ohmori T, Madoiwa S, Ozawa K, and Sakata Y.
  • Journal Title: J. Med. Virol. Volume: 86 Issue: 11 Pages: 1990-1997
  • DOI: 10.1002/jmv.23818
  • Peer Reviewed / Open Access
• [Journal Article] Manufacturing and characterization of a recombinant adeno-associated virus type 8 reference standard material. (2014)
  • Author(s): Ayuso E, et al
  • Journal Title: Hum. Gene Ther. Volume: 25 Issue: 11 Pages: 977-987
  • DOI: 10.1089/hum.2014.057
  • Peer Reviewed
• [Journal Article] Novel anti-tumor mechanism of galanin receptor type 2 in head and neck squamous cell carcinoma cells (2014)
  • Author(s): Uehara T, Kanazawa T, Mizukami H, Uchibori R, Tsukahara T, Urabe M, Kume A, Misawa K, Carey T, Suzuki M, Ichimura K, Ozawa K
  • Journal Title: Cancer Science Volume: 105 Issue: 1 Pages: 72-80
  • DOI: 10.1111/cas.12315
  • Peer Reviewed / Open Access
• [Presentation] Long-term expression of coagulation factor IX gene transduced by AAV8 into macaques (2015)
  • Author(s): Mizukami H, Ohmori T, Uchibori R, Tsukahara T, Saga, Y, Urabe M, Kume A, Sakata Y, Ozawa K
  • Organizer: 日本血液学会
  • Place of Presentation: 金沢
  • Year and Date: 2015-10-17
• [Presentation] Persistent factor IX expression following AAV8-mediated liver transduction in macaques (2015)
  • Author(s): Mizukami H, Ohmori T, Uchibori R, Tsukahara T, Saga, Y, Urabe M, Kume A, Sakata Y, Ozawa K
  • Organizer: 欧州遺伝子細胞治療学会
  • Place of Presentation: ヘルシンキ
  • Year and Date: 2015-09-17
  • Int'l Joint Research
• [Presentation] Long-term efficacy of factor IX gene expression following AAV8-mediated liver transduction in macaques (2015)
  • Author(s): Mizukami H, Ohmori T, Uchibori R, Tsukahara T, Saga, Y, Urabe M, Kume A, Sakata Y, Ozawa K.
  • Organizer: 日本遺伝子治療学会
  • Place of Presentation: 大阪
  • Year and Date: 2015-07-25
• [Presentation] Age-related eligibility of hemophilia gene therapy using AAV vectors (2014)
  • Author(s): Mizukami H, et al.
  • Organizer: The 76th Annual Meeting of the Japanese Society of Hematology
  • Place of Presentation: Osaka
  • Year and Date: 2014-11-01
• [Presentation] IMMUNE RESPONSES IN HEMOPHILIA GENE THERAPY (2014)
  • Author(s): Mizukami H
  • Organizer: The 20th Annual Meeting of Japan Society of Gene Therapy
  • Place of Presentation: Tokyo
  • Year and Date: 2014-08-07
  • Invited
• [Presentation] Low Prevalence of Neutralizing Antibody against AAV capsids in Young Japanese Population (2014)
  • Author(s): Mizukami H, Mimuro J, Tsukasa O, Madoiwa S, Sakata Y, Ozawa K
  • Organizer: 15th International Parvovirus Workshop
  • Place of Presentation: Bordeaux, France
  • Year and Date: 2014-06-25
• [Presentation] Accurate measurement of NAb status against AAV vector capsids and an approach toward managing its inhibitory effect (2013)
  • Author(s): Mizukami H, Mimuro J, Hishikawa S, Ikemoto T, Uchibori R, Tsukahara T, Urabe M, Kume A, Sakata A, Ohmori T, Madoiwa S, Sakata Y, Ozawa K
  • Organizer: 21st Annual Congress European Society for Gene and Cell Therapy
  • Place of Presentation: Madrid, Spain
• [Presentation] Precise evaluation of NAb against AAV vectors and approaches toward managing its inhibitory effect (2013)
  • Author(s): Mizukami H, Mimuro J, Uchibori R, Tsukahara T, Urabe M, Kume A, Sakata A, Ohmori T, Madoiwa S, Sakata Y, Ozawa K
  • Organizer: 第75回日本血液学会学術集会
  • Place of Presentation: 札幌
• [Book] J Thromb Haemost (2015)
  • Author(s): Ohmori T, Mizukami H, Ozawa K, Sakata Y. Nishimura S.
  • Total Pages: 10
  • Publisher: Wiley
• [Book] Gene Therapy and Cell Therapy through the Liver: Current Aspects and Future Prospects (Terai S. and Suda T. Eds) (2015)
  • Author(s): Mizukami H, Mimuro J, Ohmori T, Sakata Y, Ozawa K.
  • Total Pages: 15
  • Publisher: Springer
• [Book] Gene Therapy and Cell Therapy through the Liver: Current Aspects and Future Prospects (2015)
  • Author(s): Mizukami H, et al.
  • Total Pages: 115
  • Publisher: Springer
• [Book] 動物の事典 (2015)
  • Author(s): 水上 浩明
  • Publisher: 朝倉書店
• [Book] 遺伝子・DNA利用の製品研究開発における規制・倫理対応と解析，操作技術のトラブル対策 (2014)
  • Author(s): 水上浩明
  • Publisher: 技術情報協会