Development of a novel CRISPR-Cas9 vector for functional analysis of retinal disease-related genes

• Project/Area Number: 18K09395
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 56060:Ophthalmology-related
• Research Institution: Nagoya University (2020); Tohoku University (2018-2019)
• Principal Investigator: Fujita Kosuke 名古屋大学, 医学系研究科, 助教 (80708115)
• Co-Investigator(Kenkyū-buntansha): 中澤 徹 東北大学, 医学系研究科, 教授 (30361075) ; 西口 康二 名古屋大学, 医学系研究科, 教授 (30447825)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2020: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2019: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2018: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: アデノ随伴ウイルス / ゲノム編集 / 網膜 / 遺伝子治療 / 遺伝子編集

Outline of Final Research Achievements

Based on the miniaturization of each part required for genome editing, we developed a single AAV platform that can locally replace mutant sequences with wild-type counterparts. In blind mice, the mutation replacement rescued approximately 10% of photoreceptors, resulting in increased visual acuity to approximately 60% of controls. Surprisingly, these effects were comparable to recovery mediated by photoreceptor-targeted gene supplementation. This strategy paves the way for the treatment of hereditary disorders caused by mutations in larger genes where traditional gene replacement therapies are not currently feasible.

Academic Significance and Societal Importance of the Research Achievements

網膜色素変性は、本邦での中途失明原因の第2位の遺伝性神経変性疾患である。アデノ随伴ウイルス（AAV）を用いた遺伝子補充療法の有効性が示されているのものの、同治療の適応範囲は狭く、日本人網膜変性患者の数％しか治療対象にならない。本研究で開発したAll-in-one AAVベクターを用いたゲノム編集技術ならば、ほぼすべての変異を治療できる可能性がある。本研究により、効率的なゲノム修復ができるようになった場合、治療不可能であった遺伝子変異に対する治療が可能となる。これにより、従来では治療不可能であった遺伝子変異に対する遺伝子治療への道が拓かれた。

Research Products

• [Journal Article] In vivo imaging of the light response in mouse retinal ganglion cells based on a neuronal activity-dependent promoter (2020)
  • Author(s): Fujita Kosuke、Nishiguchi Koji M.、Sato Kota、Nakagawa Yurika、Nakazawa Toru
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 521 Issue: 2 Pages: 471-477
  • DOI: 10.1016/j.bbrc.2019.10.155
  • Peer Reviewed / Open Access
• [Journal Article] Single AAV-mediated mutation replacement genome editing in limited number of photoreceptors restores vision in mice (2020)
  • Author(s): Nishiguchi Koji M.、Fujita Kosuke、Miya Fuyuki、Katayama Shota、Nakazawa Toru
  • Journal Title: Nature Communications Volume: 11 Issue: 1 Pages: 482-482
  • DOI: 10.1038/s41467-019-14181-3
  • Peer Reviewed / Open Access