Analysis of Amyotrophic lateral sclerosis using Chmp2A / B mutant mice

• Project/Area Number: 16K07087
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Laboratory animal science
• Research Institution: Hamamatsu University School of Medicine
• Principal Investigator: Takabayashi Shuji 浜松医科大学, 光尖端医学教育研究センター, 助教 (70372521)
• Research Collaborator: AOTO kazuto
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2018: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2017: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2016: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: 筋萎縮性側索硬化症 / Chmp2A / Chmp2B / i-GONAD / マウス / ゲノム編集 / Chmp2a / Chmp2b / 筋委縮性側索硬化症 / ALS / CRISPR/CAS9 / 筋萎縮側索硬化症 / ESCRT

Outline of Final Research Achievements

We found a novel mutant mouse that spontaneously develops Amyotrophic lateral sclerosis (ALS) -like pathology and found that the causative gene is a missense mutation of the Chmp2A gene. On the other hand, its family, CHMP2B, has been reported as a causative gene for neurodegenerative diseases. Many missense mutations have been reported, particularly in human ALS patients. This study was conducted to clarify that Chmp2A is the causative gene for ALS-like abnormalities. In this study, we developed a simple gene modified mouse preparation method (i-GONAD method) to which genome editing technology was applied, thereby creating knock-in mice into which Chmp2A and Chmp2B were knockout and SNP mutations were introduced.

Academic Significance and Societal Importance of the Research Achievements

ヒトALSの原因遺伝子としていくつかが同定されている。しかし、Chmp2AがALSの原因遺伝子であるという報告はまだない。本研究でChmp2A KOマウスはChmp2B KOよりシビアな表現型を示した。Chmp2A遺伝子の遺伝子異常はヒトにおいても致死性の病態を示す可能性が示唆された。しかし、Chmp2A-L173P KIマウスでは明らかな後肢麻痺の表現型を示したことから、Chmp2Aがまだ同定されていない原因遺伝子である可能性を示唆している。本研究によりChmp2Aおよび2B遺伝子改変マウスを作製した。これらマウスの解析からALSの病態の解明につながる知見が得られる可能性がある。

Research Products

• [Journal Article] i-GONAD: a robust method for in situ germline genome engineering using CRISPR nucleases (2018)
  • Author(s): Ohtsuka Masato、Sato Masahiro、Miura Hiromi、Takabayashi Shuji、Matsuyama Makoto、Koyano Takayuki、Arifin Naomi、Nakamura Shingo、Wada Kenta、Gurumurthy Channabasavaiah B.
  • Journal Title: Genome Biology Volume: 19 Issue: 1 Pages: 1-15
  • DOI: 10.1186/s13059-018-1400-x
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] i-GONAD法を用いた遺伝子改変マウスにおける系統差 (2018)
  • Author(s): 高林 秀次、青島 拓也、椛嶋 克哉、佐藤 正宏、大塚 正人
  • Organizer: 第65回 日本実験動物学会総会