Development of CRAG gene therapy for neurodegenerative diseases

• Project/Area Number: 25640040
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Neurochemistry/Neuropharmacology
• Research Institution: Tokyo University of Pharmacy and Life Science
• Principal Investigator: YANAGI Shigeru 東京薬科大学, 生命科学部, 教授 (60252003)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000); Fiscal Year 2014: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2013: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
• Keywords: CRAG / 遺伝子治療 / 脊髄小脳変性症

Outline of Final Research Achievements

CRAG facilitated degradation of expanded polyglutamine protein (polyQ) via the nuclear ubiquitin-proteasome pathway. Taking advantage of this feature, lentivirus-mediated CRAG expression in the Purkinje cells of mice expressing polyQ resulted in clearance of the polyQ aggregates and rescue from ataxia, suggesting usefulness in targeted delivery of CRAG as a gene therapy for neurodegenerative diseases. However, a physiological relevant of CRAG in vivo are unknown. Here, we analyzed CRAG/Centaurin-γ3 KO mice. CRAG/Centaurin-γ3 KO mice spontaneously developed severe neurodegenerative phenotypes including hind-limb clasping, neuronal atrophy, cell death and lethality within 1 month of birth. Furthermore, we found that CRAG enhances neuronal cell survival against the accumulation of unfolded proteins through not only proteasome activation but also SRF-mediated c-fos activation in vivo. Our results may contribute to development of CRAG gene therapy for neurodegenerative diseases.

Research Products

• [Journal Article] Roles of mitochondrial ubiquitin ligase MITOL/MARCH5 in mitochondrial dynamics and diseases (2014)
  • Author(s): Nagashima, S., Tokuyama, T., Yonashiro, R., Inatome, R., and Yanagi S.
  • Journal Title: J. Biochem Volume: 155 Issue: 5 Pages: 273-279
  • DOI: 10.1093/jb/mvu016
  • Peer Reviewed
• [Journal Article] One-year follow-up of transgene expression by integrase-defective lentiviral vectors and their therapeutic potential in spinocerebellar ataxia model mice. (2014)
  • Author(s): Saida, H., Matsuzaki, Y., Takayama, K., Iizuka, A., Konno, A., Yanagi, S., and Hirai, H.
  • Journal Title: Gene Ther. Volume: 21 Pages: 820-827
  • Peer Reviewed
• [Journal Article] Downregulation of Centaurin gamma1A increases synaptic transmission at Drosophila larval neuromuscular junctions (2014)
  • Author(s): Homma, M.、 Nagashima, S., Fukuda, T., Yanagi, S., Miyakawa, H., Suzuki, E., Morimoto, T.
  • Journal Title: Europian Journal of Neuroscience Volume: VOL40 Issue: 8 Pages: 3158-3170
  • DOI: 10.1111/ejn.12681
  • Peer Reviewed
• [Journal Article] Mutant ataxin-3 with an abnormally expanded polyglutamine chain disrupts dendritic development and metabotropic glutamate receptor signaling in mouse cerebellar purkinje cells (2014)
  • Author(s): Konno A, Shuvaev AN, Miyake N, Miyake K, Iizuka A, Matsuura S, Huda F, Nakamura K, Yanagi S, Shimada T, Hirai H.
  • Journal Title: Cerebellum Volume: 13 Issue: 1 Pages: 29-41
  • DOI: 10.1007/s12311-013-0516-5
  • Peer Reviewed
• [Journal Article] MITOLによるミトコンドリアダイナミクス制御と破綻による疾患 (2014)
  • Author(s): 長島 駿、柳 茂
  • Journal Title: 生化学 Volume: 86 Pages: 63-67
  • NAID: 40020011079
  • Peer Reviewed
• [Journal Article] Simultaneous measurement of cytosolic and mitochondrial Ca(2+) during ischemia in mice whole-brain slice preparation and its application to drug evaluation. (2013)
  • Author(s): Nishiyama, T., Hasegawa, E., Yanagi, S., Kudo, Y., Hamada, R., Matsumura, N., Tomino, M., Muromachi, Y., Hatakeyama, K., and Uchino, H.
  • Journal Title: Acta Neurochir. Suppl. Volume: 118 Pages: 65-70
  • DOI: 10.1007/978-3-7091-1434-6_11
  • ISBN: 9783709114339, 9783709114346
  • Peer Reviewed
• [Journal Article] MITOL regulates endoplasmic reticulum-mitochondria contacts via Mitofusin2. (2013)
  • Author(s): Sugiura, A., Nagashima, S., Tokuyama, T., Amo, T., Matsuki, Y., Ishido, S., Kudo, Y., McBride, H.M., Fukuda, T., Matsushita, T., Inatome, R., and C Yanagi, S.
  • Journal Title: Mol. Cell Volume: 51 Issue: 1 Pages: 1-15
  • DOI: 10.1016/j.molcel.2013.04.023
  • Peer Reviewed
• [Presentation] CRAGによるSRF活性化を介した神経細胞の生存シグナル機構 (2014)
  • Author(s): 柳 茂、福田 敏史、稲留 涼子、長島 駿
  • Organizer: 第37回日本神経科学会大会
  • Place of Presentation: 横浜
  • Year and Date: 2014-09-11
• [Presentation] Role of MITOL in mitochondrial dynamics
  • Author(s): Shun Nagashima, Ayumu Sugiura, Shigeru Yanagi
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: 神戸
• [Presentation] TAX1BP1 plays a role in immunoglobulin gene diversification through regulation of somatic hyper mutation
  • Author(s): 松下暢子, 鈴木みどり, 柳 茂
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: 神戸
• [Presentation] ミトコンドリア新機能と疾患
  • Author(s): 武田 啓佑, 長島 駿, 柳 茂
  • Organizer: 東京医科大学第172回医学会総会
  • Place of Presentation: 東京
• [Presentation] TAX1BP1は体細胞高頻度突然変異を制御することによって抗体遺伝子多様化機構に機能する
  • Author(s): 松下暢子, 鈴木みどり, 伊波英克, 柳 茂
  • Organizer: 第72回日本癌学会学術総会
  • Place of Presentation: 横浜
• [Presentation] TAX1BP1 regulates IgM expression at the cell surface through NF-κB pathway
  • Author(s): 鈴木みどり, 松下暢子, 伊波英克, 柳 茂
  • Organizer: 第72回日本癌学会学術総会
  • Place of Presentation: 横浜
• [Presentation] Role of mitochondrial ubiquitin ligase MITOL in mitochondrial dynamics and diseases
  • Author(s): 長島 駿, 杉浦 歩, 徳山剛士, 武田啓佑, 稲留涼子, 柳 茂
  • Organizer: 第86回日本生化学会大会
  • Place of Presentation: 横浜