Analysis of the molecular link of amyotrophic lateral sclerosis associated genes in sporadic ALS model mice

• Project/Area Number: 25461269
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Neurology
• Research Institution: The University of Tokyo
• Principal Investigator: Yamashita Takenari 東京大学, 医学(系)研究科(研究院), 特任研究員 (20431843)
• Co-Investigator(Renkei-kenkyūsha): KWAK Shin 東京大学, 大学院医学系研究科, 客員研究員 (40160981)
• 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: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2014: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2013: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: 筋萎縮性側索硬化症 / カルパイン / RNA編集 / カルシウム / TDP-43 / ADAR2 / AMPA受容体 / リン酸化 / calpain / 神経細胞死 / RNA結合蛋白 / FUS / GluA2

Outline of Final Research Achievements

We found that ADAR2-negative motor neurons in the spinal cord showed TDP-43 pathology, or subcellular mislocalization of TDP-43, in the conditional ADAR2 knockout (AR2) mice, a mechanistic ALS model in which the ADAR2 gene was targeted in cholinergic neurons including motor neurons. TDP-43 pathology in the ADAR2-lacking motor neurons in the AR2 mice was caused by abnormal activation of calpain, a Ca2+-dependent cysteine protease. We revealed several calpain-dependent cleavage sites in the C-terminal region of TDP-43. Moreover, we found that phosphorylated full-length TDP-43 and calpain-dependent TDP-43 fragments were more resistant to cleavage by calpain than endogenous full-length TDP-43 was.

Research Products

• [Journal Article] Deficient RNA-editing enzyme ADAR2 in an amyotrophic lateral sclerosis patient with a FUSP525L mutation. (2016)
  • Author(s): Aizawa H, Hideyama T, Yamashita T, Kimura T, Suzuki N, Aoki M, Kwak S
  • Journal Title: J Clin Neurosci Volume: -
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Phosphorylated TDP-43 becomes resistant to cleavage by calpain: a regulatory role for phosphorylation in TDP-43 pathology of ALS/FTLD. (2016)
  • Author(s): Yamashita T, Teramoto S, Kwak S
  • Journal Title: Neurosci Res Volume: 未定 Pages: 63-69
  • DOI: 10.1016/j.neures.2015.12.006
  • Peer Reviewed / Open Access
• [Journal Article] Autophagy in spinal motor neurons of conditional ADAR2-knockout mice: an implication for a role of calcium in increased autophagy flux in ALS. (2015)
  • Author(s): Sasaki S, Yamashita T, Kwak S
  • Journal Title: Neurosci Lett Volume: 598 Pages: 79-84
  • DOI: 10.1016/j.neulet.2015.05.025
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Unique nuclear vacuoles in the motor neurons of conditional ADAR2-knockout mice (2014)
  • Author(s): Sasaki S, Yamashita T, Hideyama T, Kwak S
  • Journal Title: Brain Research Volume: 1550 Pages: 36-46
  • DOI: 10.1016/j.brainres.2014.01.006
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] The molecular link between inefficient GluA2 Q/R site-RNA editing and TDP-43 pathology in motor neurons of sporadic amyotrophic lateral sclerosis patients. (2014)
  • Author(s): Yamashita, T., and Kwak, S.
  • Journal Title: Brain Res. Volume: in press. Pages: 28-38
  • DOI: 10.1016/j.brainres.2013.12.011
  • Peer Reviewed / Open Access
• [Journal Article] TDP-43病理形成メカニズム (2014)
  • Author(s): 山下雄也, 郭 伸
  • Journal Title: Dementia Japan Volume: 28 Pages: 307-318
• [Journal Article] Calpain plays a crucial role in TDP-43 pathology (2014)
  • Author(s): 山下雄也, 郭 伸
  • Journal Title: Rinsho Shinkeigaku Volume: 54 Issue: 12 Pages: 1151-1154
  • DOI: 10.5692/clinicalneurol.54.1151
  • NAID: 130004921169
  • ISSN: 0009-918X, 1882-0654
• [Journal Article] Rescue of amyotrophic lateral sclerosis phenotype in a mouse model by intravenous AAV9-ADAR2 delivery to motor neurons (2013)
  • Author(s): Yamashita T, Chai HL, Teramoto S, Tsuji S, Shimazaki K, Muramatsu S, Kwak S
  • Journal Title: EMBO Mol Med Volume: 5(11) Issue: 11 Pages: 1710-1719
  • DOI: 10.1002/emmm.201302935
  • Peer Reviewed
• [Journal Article] A unique mouse model for investigating the properties of amyotrophic lateral sclerosis-associated protein TDP-43, by in utero electroporation. (2013)
  • Author(s): Akamatsu, M., Takuma, H., Yamashita, T., Okada, T., Keino-Masu, K., Ishii, K., Kwak, S., Masu, M., and Tamaoka, A.
  • Journal Title: Neuroscience Research Volume: 77 Issue: 4 Pages: 234-241
  • DOI: 10.1016/j.neures.2013.09.009
  • NAID: 120007130577
  • Peer Reviewed
• [Journal Article] RNA editingと筋萎縮性側索硬化症。RNA Biologyからみた神経変性疾患の病態機序 (2013)
  • Author(s): 山下雄也、郭 伸
  • Journal Title: 医学の歩み Volume: 247 Pages: 412-420
• [Presentation] AMPA受容体アンタゴニストであるランパネルを用いたALS治療法の開発 (2015)
  • Author(s): 山下雄也, 廣瀬直己, 赤松恵, 寺本さやか, 蔡慧令, 郭伸
  • Organizer: 第38回日本神経科学大会
  • Place of Presentation: 神戸国際会議場.兵庫県・神戸市
  • Year and Date: 2015-07-28
• [Presentation] Mechanism-based gene therapy for sporadic ALS. (2015)
  • Author(s): 郭 伸, 山下雄也, 寺本さやか
  • Organizer: 第56回日本神経学会学術大会
  • Place of Presentation: 朱鷺メッセ(新潟コンベンションセンター). 新潟県・新潟市
  • Year and Date: 2015-05-20
• [Presentation] Gene therapy for sporadic ALS using an intravenous injection of AAV vector. (2014)
  • Author(s): Yamashita T, Chai HL, Teramoto S, Muramatsu S, Kwak S
  • Organizer: The 25th International Symposium on MND/ALS
  • Place of Presentation: Brussels, Belgium
  • Year and Date: 2014-12-05 – 2014-12-07
• [Presentation] Mechanism-based gene therapy for ALS using sporadic ALS model mice (2014)
  • Author(s): Yamashita T, Chai HL, Teramoto S, Tsuji S, Shimazaki K, Muramatsu S-I, Kwak S
  • Organizer: The 45th Annual Meeting Society for Neuroscience
  • Place of Presentation: Washington DC, USA
  • Year and Date: 2014-11-15 – 2014-11-19
• [Presentation] Electron microscopic observation of intranuclear aggregation of TDP-43 in mouse cerebral cortex produced by in utero electroporation (2014)
  • Author(s): Akamatsu M, Takuma H, Yamashita T, Okada T, Masu-Keino K, Oehring H, Kwak S, Masu M, Jirikowski FG
  • Organizer: 第37回日本神経科学大会Neuro2014
  • Place of Presentation: パシフィコ横浜, 横浜、日本
  • Year and Date: 2014-09-11 – 2014-09-13
• [Presentation] ALSモデルマウスを用いたALSの遺伝子治療法開発の試み. (2014)
  • Author(s): 山下雄也, 蔡 慧玲, 寺本さやか, 辻 省次, 島崎久仁子, 村松慎一, 郭 伸
  • Organizer: 第37回日本神経科学大会Neuro2014
  • Place of Presentation: パシフィコ横浜, 横浜、日本
  • Year and Date: 2014-09-11 – 2014-09-13
• [Presentation] ペランパネルはALSモデルマウス運動ニューロンのTDP-43病理を軽減する (2014)
  • Author(s): 蔡 慧玲, 郭 伸, 山下雄也
  • Organizer: 第37回日本神経科学大会Neuro2014
  • Place of Presentation: パシフィコ横浜, 横浜、日本
  • Year and Date: 2014-09-11 – 2014-09-13
• [Presentation] Development of a gene therapy for sporadic ALS by normalizing ADAR2 activity in the motor neurons using a vascular type AAV vector. (2014)
  • Author(s): Kwak S, Yamashita T, Chai HL, Teramoto S, Muramatsu S-I
  • Organizer: The 24th Meeting of the European Neurological Society
  • Place of Presentation: Istanbul, Turkey
  • Year and Date: 2014-05-31 – 2014-06-03
• [Presentation] 分子病態に基づいた孤発性ALSの遺伝子治療法の開発 (2014)
  • Author(s): 郭 伸, 山下雄也, 蔡 慧玲, 寺本さやか, 辻省次, 島崎久仁子, 村松愼一
  • Organizer: 第55回日本神経学会学術大会
  • Place of Presentation: 福岡国際会議場, 福岡, 日本
  • Year and Date: 2014-05-21 – 2014-05-24
• [Presentation] 子宮内電気穿孔法TDP-43遺伝子導入によるin vivo形成封入体の微細形態の検討 (2014)
  • Author(s): 詫間 浩, 赤松 恵, 山下雄也, Hartmut Oehring, 岡田拓也, 枡 和子, 石井一弘, Gustav Jirikowsk, 郭 伸, 枡 正幸, 玉岡 晃
  • Organizer: 第55回日本神経学会学術大会
  • Place of Presentation: 福岡国際会議場, 福岡, 日本
  • Year and Date: 2014-05-21 – 2014-05-24
• [Presentation] AMPA受容体サブユニットGluA2のQ/R部位RNA編集率に対する薬剤の効果 (2014)
  • Author(s): 澤田 潤, 相澤仁志, 浅野目明日香, 高橋佳恵, 斎藤 司, 片山隆行, 山下雄也, 郭 伸, 長谷部直幸
  • Organizer: 第55回日本神経学会学術大会
  • Place of Presentation: 福岡国際会議場, 福岡, 日本
  • Year and Date: 2014-05-21 – 2014-05-24
• [Presentation] 筋炎と悪性腫瘍との関連に関する疫学的検討 (2014)
  • Author(s): 肥田あゆみ, 井上真奈美, 前田明子, 山下雄也, 郭 伸、辻 省次, 清水 潤
  • Organizer: 第55回日本神経学会学術大会
  • Place of Presentation: 福岡国際会議場, 福岡, 日本
  • Year and Date: 2014-05-21 – 2014-05-24
• [Presentation] TDP-43病理形成メカニズムにおけるTDP-43のカルパイン依存性断片化の意義 (2014)
  • Author(s): 山下雄也, 郭 伸
  • Organizer: 第55回日本神経学会学術大会
  • Place of Presentation: 福岡国際会議場, 福岡, 日本
  • Year and Date: 2014-05-21 – 2014-05-24
  • Invited
• [Presentation] Molecular mechanism generating TDP-43 pathology in amyotrophic lateral sclerosis (ALS) motor neurones. (2013)
  • Author(s): Kwak S, Yamashita T, Hideyama T, Teramoto S
  • Organizer: The 23rd Meeting of the European Neurological Society
  • Place of Presentation: Balcerona, Spain
• [Presentation] ADAR2-dependent death and TDP-43 mislocalization in motor neurons. (2013)
  • Author(s): Kwak S, Yamashita T, Teramoto S, Chai HL
  • Organizer: 8th Brain Research conference “RNA binding proteins in neurological disease”
  • Place of Presentation: San Diego, USA
• [Presentation] Calpain-dependent cleavage of TDP-43 plays a crucial role in ALS pathology (2013)
  • Author(s): Yamashita T, Hideyama T, Teramoto S, Takano J, Iwata N, Saido TC, Kwak S
  • Organizer: 8th Brain Research conference “RNA binding proteins in neurological disease”
  • Place of Presentation: San Diego, USA
• [Presentation] Calpain-dependent cleavage of TDP-43 plays a crucial role in ALS pathology. (2013)
  • Author(s): 4. Yamashita T, Hideyama T, Teramoto S, Takano J, Iwata N, Saido TC, Kwak S
  • Organizer: The 44th Annual Meeting Society for Neuroscience
  • Place of Presentation: San Diego, USA
• [Presentation] ADAR2 plays a key role for death and TDP-43 mislocalization in ALS motor neurons. (2013)
  • Author(s): Yamashita T, Teramoto S, Chai HL, Muramatsu SI, Kwak S
  • Organizer: The 24th International Symposium on MND/ALS
  • Place of Presentation: Milan, Itary
• [Presentation] AMPA受容体サブユニットGluA2のQ/R 部位RNA編集率に対する薬剤の効果 (2013)
  • Author(s): 澤田準、相澤仁志、淺野目明日香、高橋佳恵、斉藤司、片山隆行、長谷部直幸、山下雄也、郭 伸
  • Organizer: 第54回日本神経学会学術大会
  • Place of Presentation: 東京、日本
• [Presentation] ClapainがALSのTDP-43病理形成に重要な役割を果たす (2013)
  • Author(s): 7. 山下雄也、日出山拓人、寺本さやか、高野二郎、岩田修永、西道隆臣、郭 伸
  • Organizer: 第36回日本神経科学大会Neuro2013
  • Place of Presentation: 京都、日本
• [Presentation] ClapainがALSのTDP-43病理形成に果たす役割 (2013)
  • Author(s): 8. 山下雄也、日出山拓人、寺本さやか、高野二郎、岩田修永、西道隆臣、郭 伸
  • Organizer: 第35回神経組織培養研究会
  • Place of Presentation: 大阪、日本
• [Presentation] TDP-43 aggregation produced by In utero electroporation and its properties in the mouse motor neurons. (2013)
  • Author(s): 9. Akamatsu M, Takuma H, Yamashita T, Okada T, Ishii K, Keino-Masu K, Kwak S, Masu M, Tamaoka A
  • Organizer: The 44th Annual Meeting Society for Neuroscience
  • Place of Presentation: San Diego、USA
• [Presentation] 筋萎縮性側索硬化症関連タンパク質：TDP-43封入体の形成ーマウス胎仔電気穿孔法による大脳皮質運動野への疾患関連遺伝子の導入 (2013)
  • Author(s): 10. 赤松恵、詫間浩、山下雄也、岡田拓也、石井一弘、桝和子、郭 伸、桝正幸、玉岡晃
  • Organizer: 第36回日本神経科学大会Neuro2013
  • Place of Presentation: 京都、日本
• [Presentation] マウス胎仔電気穿孔法による in vivo TDP-43 遺伝子導入と封入体形成 (2013)
  • Author(s): 11. 詫間浩、赤松恵、山下雄也、岡田拓也、石井一弘、桝和子、郭 伸、桝正幸、玉岡晃
  • Organizer: 第54回日本神経学会学術大会
  • Place of Presentation: 東京、日本
• [Book] 遺伝子医学MOOK 30号 (2016)
  • Author(s): 山下雄也, 郭 伸
  • Publisher: 株式会社 メディカル ドゥ
• [Remarks] 東京大学大学院医学系研究科 郭 研究室
  • URL: http://square.umin.ac.jp/teamkwak/
• [Remarks] 東京大学大学院医学系研究科 疾患生命工学センター 臨床医工学部門 郭 研究室
  • URL: http://square.umin.ac.jp/teamkwak/
• [Remarks] マウスにおける筋萎縮性側索硬化症の遺伝子治療実験に成功
  • URL: http://www.u-tokyo.ac.jp/ja/utokyo-research/research-news/a-major-step-towards-the-cure-of-sporadic-als/
• [Remarks] マウスにおける筋萎縮性側索硬化症の遺伝子治療実験に成功
  • URL: http://www.m.u-tokyo.ac.jp/news/press.html#20130926
• [Remarks] 東京大学大学院医学系研究科 疾患生命工学センター 臨床医工学部門 郭 研究室
  • URL: http://square.umin.ac.jp/teamkwak/