Development a novel gene therapy for Lysosomal disease with neurological disorders

• Project/Area Number: 15K09604
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Pediatrics
• Research Institution: Nippon Medical School
• Principal Investigator: MIYAKE NORIKO 日本医科大学, 医学部, テクニカルスタッフ (00421206)
• Co-Investigator(Kenkyū-buntansha): 三宅 弘一 日本医科大学, 医学部, 准教授 (90267211)
• Project Period (FY): 2015-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2017: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2016: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2015: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: 遺伝子治療 / 異染性白質ジストロフィー / アデノ随伴ウイルス / リソゾーム病 / 脳神経病変 / アデノ随伴ウイルスベクター / 血液脳関門 / AAV vector

Outline of Final Research Achievements

Metachromatic leukodystrophy (MLD) is a lysosomal storage disease caused by the deficiency of arylsulfatase A (ASA) and characterized by neurological symptoms. To treat adult MLD mice, we generated self-complementary type 9 AAV vector expressing ASA (scAAV9/ASA), which could cross the BBB, and examined the feasibility of scAAV9/ASA mediated gene therapy for MLD. After scAAV9/ASA injection, immunohistochemical analysis showed efficient ASA expression was detected in systemic organs and brain. Alcian blue staining and quantative analysis of sulfatide showed decrease of the amount of stored sulfatide in scAAV9/ASA treated MLD mouse. In the behavior test, scAAV9/ASA treated mice showed a significant improvement in their ability to traverse narrow balance beams as compared to non-treated MLD mice. These data indicate that IV injection of scAAV9/ASA is effective for suppression of sulfatide storage in brain and this therapeutic approach may be useful for gene therapy of adult MLD patients.

Academic Significance and Societal Importance of the Research Achievements

本研究では脳全体の広範な神経変性を伴う、異染性白質ジストロフィーをモデルとし、非侵襲的かつ安全で脳神経組織に長期の酵素補充療法が出来る治療法の開発を行った。
self-complementary AAV 9型を用いることで、脳組織の治療が可能になり、リソゾーム病のみならず、ALS、アルツハイマー病などの様々な脳神経病変を伴う疾患に応用が可能なると示唆された。

Research Products

• [Journal Article] Improved Intravitreal AAV-Mediated Inner Retinal Gene Transduction after Surgical Internal Limiting Membrane Peeling in Cynomolgus Monkeys. (2017)
  • Author(s): Takahashi K, Igarashi T, Miyake K, Kobayashi M, Yaguchi C, Iijima O, Yamazaki Y, Katakai Y, Miyake N, Kameya S, Shimada T, Takahashi H, Okada T.
  • Journal Title: Mol Ther. Volume: 4；25 Issue: 1 Pages: 296-302
  • DOI: 10.1016/j.ymthe.2016.10.008
  • Peer Reviewed / Open Access
• [Journal Article] D816V mutation in the KIT gene activation loop has greater cell-proliferative and anti-apoptotic ability than N822K mutation in core-binding factor acute myeloid leukemia (2017)
  • Author(s): Omori Ikuko、Yamaguchi Hiroki、Miyake Koichi、Miyake Noriko、Kitano Tomoaki、Inokuchi Koiti
  • Journal Title: Experimental Hematology Volume: 52 Pages: 56-64.e4
  • DOI: 10.1016/j.exphem.2017.05.003
  • Peer Reviewed
• [Journal Article] MicroRNA cluster miR-17-92 regulates multiple functionally related voltage-gated potassium channels in chronic neuropathic pain (2017)
  • Author(s): Sakai Atsushi、Saitow Fumihito、Maruyama Motoyo、Miyake Noriko、Miyake Koichi、Shimada Takashi、Okada Takashi、Suzuki Hidenori
  • Journal Title: Nature Communications Volume: 8 Issue: 1 Pages: 16079-16079
  • DOI: 10.1038/ncomms16079
  • Peer Reviewed / Open Access
• [Journal Article] Tyrosine triple mutated AAV2-BDNF gene therapy in a rat model of transient IOP elevation (2016)
  • Author(s): Igarashi T, Miyake K, Kobayashi M, Kameya S, Fujimoto C, Nakamoto K, Takahashi H, Igarashi T, Miyake N, Iijima O, Hirai Y, Shimada T, Okada T, Takahashi H.
  • Journal Title: Mol Vis. Volume: 16 Pages: 816-826
  • Peer Reviewed
• [Journal Article] Prevention of Lethal Murine Hypophosphatasia by Neonatal Ex Vivo Gene Therapy Using Lentivirally Transduced Bone Marrow Cells. (2015)
  • Author(s): Iijima O, Miyake K, Watanabe A, Miyake N, Igarashi T, Kanokoda C, Nakamura-Takahashi A, Kinoshita H, Noguchi T, Abe S, Narisawa S, Millán JL, Okada T, Shimada T.
  • Journal Title: Hum Gene Ther. Volume: 26 Issue: 12 Pages: 801-12
  • DOI: 10.1089/hum.2015.078
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Enzyme replacement in the CSF to treat metachromatic leukodystrophy in mouse model using single intracerebroventricular injection of self-complementary AAV1 vector. (2015)
  • Author(s): Hironaka K, Yamazaki Y, Hirai Y, Yamamoto M, Miyake N, Miyake K, Okada T, Morita A, Shimada T.
  • Journal Title: Sci Rep. Volume: 18 Issue: 1 Pages: 1-12
  • DOI: 10.1038/srep13104
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] Direct comparison between single-stranded and self-complementary type 9 AAV vector to treat adult MLD model mice by intravenous injection (2018)
  • Author(s): Noriko Miyake, Koichi Miyake, Motoko Yamamoto, Takashi Shimada, Takashi Okada
  • Organizer: The 24th Annual Meeting of Japan Society of Gene Therapy
• [Presentation] Intravitreal injection of AAV vector in cynomolgus monkeys affects neutralizing antibody titer against AAV in the serum (2018)
  • Author(s): Tsutomu Igarashi, Kazuhisa Takahashi, Koichi Miyake, Maika Kobayashi, Chiemi Yaguchi, Noriko Miyake, Shuhei Kameya, Hiroshi Takahashi, Takashi Okada
  • Organizer: The 24th Annual Meeting of Japan Society of Gene Therapy
• [Presentation] Why are scAAV9 vectors able to transduce the CNS but not ssAAV9? ; The difference between the ssAAV9 and scAAV9 vector in transduction of CNS by intravenous injection. (2017)
  • Author(s): Noriko Miyake, Koichi Miyake, Motoko Yamamoto, Takashi Shimada, Takashi Okada
  • Organizer: The 23th Annual Meeting of Japan Society of Gene Therapy
• [Presentation] Why are scAAV9 vectors able to pass through the BBB but not ssAAV9? ; The difference between the ssAAV9 and scAAV9 vector in transduction of CNS by intravenous injection. (2017)
  • Author(s): Noriko Miyake, Koichi Miyake, Motoko Yamamoto, Takashi Shimada, Takashi Okada
  • Organizer: 20th Annual Meeting of the American Society of Gene & Cell Therapy
  • Int'l Joint Research
• [Presentation] ssAAV vectors are able to pass through the BBB as same as dsAAV (2016)
  • Author(s): Noriko Miyake, Koichi Miyake, Motoko Yamamoto, Takashi Shimada, Takashi Okada
  • Organizer: The 22th Annual Meeting of Japan Society of Gene Therapy
  • Place of Presentation: 東京
  • Year and Date: 2016-07-28
• [Presentation] Successful treatment of neonatal metachromatic leukodystrophy model mice by low dose of self-complementary AAV type 9 vector expressing ASA (2016)
  • Author(s): Miyake N, Miyake K, Yamamoto M, Shimada T, Okada T
  • Organizer: 19th Annual Meeting of the American Society of Gene & Cell Therapy
  • Place of Presentation: Washington DC)
  • Year and Date: 2016-05-04
  • Int'l Joint Research
• [Presentation] アデノ随伴ウイルスベクターの脳室内注入による異染性白色ジストロフィーモデルマウスの遺伝子治療 (2015)
  • Author(s): 廣中浩平,山崎吉之,平井幸彦, 山本基子, 三宅紀子, 三宅弘一, 森田明夫, 島田隆, 岡田尚巳
  • Organizer: 第20 回 日本ライソゾーム病研究会
  • Place of Presentation: 東京(慈恵医科大学）
  • Year and Date: 2015-10-02
• [Presentation] 異染性白質ジストロフィー(MLD)の遺伝子治療基盤研究 (2015)
  • Author(s): 三宅 紀子、三宅 弘一、山本 基子、島田 隆、岡田 尚巳
  • Organizer: 第20 回 日本ライソゾーム病研究会
  • Place of Presentation: 東京(慈恵医科大学）
  • Year and Date: 2015-10-02
• [Presentation] Successful treatment of neonatal metachromatic leukodystrophy model mice by intravenous injection of self-complementary AAV type 9 vector expressing ASA (2015)
  • Author(s): Noriko Miyake, Koichi Miyake, Motoko Yamamoto, Takashi Shimada, Takashi Okada
  • Organizer: The 21th Annual Meeting of Japan Society of Gene Therapy
  • Place of Presentation: 大阪（国際会議場）
  • Year and Date: 2015-07-24
• [Book] Neonatal Gene Therapy for Inherited Disorders. Selected Topics in Neonatal Care. (2018)
  • Author(s): Miyake K, Miyake N, Shimada T.
  • Total Pages: 9
  • Publisher: InTech
• [Book] Gene Therapy - Principles and Challenges (2015)
  • Author(s): Koichi Miyake, Noriko Miyake and Takashi Shimada
  • Total Pages: 208
  • Publisher: INTEC