Elucidation for pathomechanisms of amyotrophic lateral sclerosis using genetically modified models.

• Project/Area Number: 20K07887
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 52020:Neurology-related
• Research Institution: Hiroshima University
• Principal Investigator: MARUYAMA Hirofumi 広島大学, 医系科学研究科(医), 教授 (90304443)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2022: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2021: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2020: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: オプチニューリン / 筋萎縮性側索硬化症 / モデル動物 / 遺伝子改変

Outline of Research at the Start

筋萎縮性側索硬化症 (Amyotrophic lateral sclerosis: ALS)の原因遺伝子であるオプチニューリン（optineurin: OPTN)遺伝子改変マウス・ノックダウン細胞を用いて生化学的・形態学的変化を検討し、ALSの発症機序を解明する。OPTN遺伝子改変によって影響が想定される神経炎症・ユビキチン系およびオートファジー経路における治療候補薬を探索する。

Outline of Final Research Achievements

We generated Optineurin knockout (OPTN KO) mice. Anterior horn neurons decreased. OPTN KO mice presented abnormal neuronal autophagic vacuoles, and showed TDP43 positive cytoplastic inclusions. We could not observe significant differences in survival time, body weight, and motor functions. In model cells, OPTN knockdown inhibited myogenic differentiation accompanied by reduction of mRNA and protein expression levels of myogenin and MyoD. OPTN may play a role in early stage of muscle differentiation. Model mice and model cells of OPTN could reproduce pathophysiology of amyotrophic lateral sclerosis (ALS).
We evaluated histopathology of peripheral axons in the skeletal muscle of human. Axonal phosphorylated TDP43 accumulated from early stage of ALS. TDP43 in nerve bundles may be a novel diagnostic biomarker for ALS.

Academic Significance and Societal Importance of the Research Achievements

オプチニューリンのモデルマウス・モデル細胞は筋萎縮性側索硬化症の病態の一部を再現しており、病態モデルとして使用できることが明らかとなった。筋萎縮性側索硬化症発症メカニズムにおけるオプチニューリンの意義を解明できたことにより、治療介入の手がかりを得ることができた。
中枢神経からの組織を生前に得ることは困難であるのに対し、筋生検での筋内神経束のリン酸化TDP43蓄積は検体採取による生前の評価が可能であり、かつ病気の早期から出現していることより、筋萎縮性側索硬化症のバイオマーカーとして早期診断の手がかりが得られた。また発症に至る病態を考える上で、運動神経終末からの逆行性進展機序を示唆する知見と言える。

Research Products

• [Journal Article] 遺伝性ALSオーバービュー (2023)
  • Author(s): 森野豊之、丸山博文
  • Journal Title: Clinical Neuroscience Volume: 41 Pages: 330-333
• [Journal Article] Efficacy and Safety of Ultrahigh-Dose Methylcobalamin in Early-Stage Amyotrophic Lateral Sclerosis (2022)
  • Author(s): Oki Ryosuke et al.
  • Journal Title: JAMA Neurology Volume: 79 Issue: 6 Pages: 575-575
  • DOI: 10.1001/jamaneurol.2022.0901
  • Peer Reviewed / Open Access
• [Journal Article] TDP-43 Accumulation Within Intramuscular Nerve Bundles of Patients With Amyotrophic Lateral Sclerosis (2022)
  • Author(s): Kurashige Takashi、Morino Hiroyuki、Murao Tomomi、Izumi Yuishin、Sugiura Tomohito、Kuraoka Kazuya、Kawakami Hideshi、Torii Tsuyoshi、Maruyama Hirofumi
  • Journal Title: JAMA Neurology Volume: 79 Issue: 7 Pages: 693-693
  • DOI: 10.1001/jamaneurol.2022.1113
  • Peer Reviewed / Open Access
• [Journal Article] Detecting motor unit abnormalities in amyotrophic lateral sclerosis using high-density surface EMG (2022)
  • Author(s): Nishikawa Yuichi、Holobar Ales、Watanabe Kohei、Takahashi Tetsuya、Ueno Hiroki、Maeda Noriaki、Maruyama Hirofumi、Tanaka Shinobu、Hyngstrom Allison S.
  • Journal Title: Clinical Neurophysiology Volume: 142 Pages: 262-272
  • DOI: 10.1016/j.clinph.2022.06.016
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Knockdown of optineurin controls C2C12 myoblast differentiation via regulating myogenin and MyoD expressions (2022)
  • Author(s): Ishikawa Kenichi、Araki Mutsuko、Nagano Yoshito、Motoda Atsuko、Shishido Takeo、Kurashige Takashi、Takahashi Tetsuya、Morino Hiroyuki、Kawakami Hideshi、Matsumoto Masayasu、Maruyama Hirofumi
  • Journal Title: Differentiation Volume: 123 Pages: 1-8
  • DOI: 10.1016/j.diff.2021.11.004
  • Peer Reviewed / Open Access
• [Journal Article] An autopsy report of a familial amyotrophic lateral sclerosis case carrying <scp> <i>VCP</i> Arg487His </scp> mutation with a unique <scp>TDP‐43</scp> proteinopathy (2021)
  • Author(s): Matsubara Tomoyasu、Izumi Yuishin、Oda Masaya、Takahashi Masatoshi、Maruyama Hirofumi、Miyamoto Ryosuke、Watanabe Chigusa、Tachiyama Yoshiro、Morino Hiroyuki、Kawakami Hideshi、Saito Yuko、Murayama Shigeo
  • Journal Title: Neuropathology Volume: 41 Issue: 2 Pages: 118-126
  • DOI: 10.1111/neup.12710
  • Peer Reviewed / Open Access
• [Journal Article] Case of Neuronal Intranuclear Inclusion Disease With Dynamic Perfusion Changes Lacking Typical Signs on Diffusion-Weighted Imaging (2021)
  • Author(s): Kikumoto Mai、Nezu Tomohisa、Shiga Yuji、Motoda Atsuko、Toko Megumi、Kurashige Takashi、Ueno Hiroki、Takahashi Tetsuya、Morino Hiroyuki、Sone Jun、Iwasaki Yasushi、Sobue Gen、Maruyama Hirofumi
  • Journal Title: Neurology Genetics Volume: 7 Issue: 4
  • DOI: 10.1212/nxg.0000000000000601
  • Peer Reviewed / Open Access
• [Journal Article] An autopsied case of ADSSL1 myopathy (2021)
  • Author(s): Motoda Atsuko、Takahashi Tetsuya、Watanabe Chigusa、Tachiyama Yoshiro、Ochi Kazuhide、Saito Yoshihiko、Iida Aritoshi、Nishino Ichizo、Maruyama Hirofumi
  • Journal Title: Neuromuscular Disorders Volume: 31 Issue: 11 Pages: 1220-1225
  • DOI: 10.1016/j.nmd.2021.07.011
  • Peer Reviewed / Open Access
• [Journal Article] Optineurin defects cause TDP43-pathology with autophagic vacuolar formation (2021)
  • Author(s): Kurashige Takashi、Kuramochi Masahito、Ohsawa Ryosuke、Yamashita Yui、Shioi Go、Morino Hiroyuki、Kamada Masaki、Ayaki Takashi、Ito Hidefumi、Sotomaru Yusuke、Maruyama Hirofumi、Kawakami Hideshi
  • Journal Title: Neurobiology of Disease Volume: 148 Pages: 105215-105215
  • DOI: 10.1016/j.nbd.2020.105215
  • Peer Reviewed / Open Access
• [Journal Article] Anti-HMGCR Antibody-Positive Myopathy Shows Bcl-2-Positive Inflammation and Lymphocytic Accumulations (2020)
  • Author(s): Kurashige Takashi、Murao Tomomi、Mine Naoko、Sugiura Tomohito、Inazuka Yukiko、Kuraoka Kazuya、Takahashi Tetsuya、Maruyama Hirofumi、Torii Tsuyoshi
  • Journal Title: Journal of Neuropathology & Experimental Neurology Volume: 79 Issue: 4 Pages: 448-457
  • DOI: 10.1093/jnen/nlaa006
  • Peer Reviewed
• [Journal Article] Nicotine-induced upregulation of miR-132-5p enhances cell survival in PC12 cells by targeting the anti-apoptotic protein Bcl-2 (2020)
  • Author(s): Shrestha Tejashwi、Takahashi Tetsuya、Li Chengyu、Matsumoto Masayasu、Maruyama Hirofumi
  • Journal Title: Neurological Research Volume: 42 Issue: 5 Pages: 405-414
  • DOI: 10.1080/01616412.2020.1735817
  • Peer Reviewed
• [Presentation] TDP43 accumulates in itramuscular nerve bundles of ALS patients. (2022)
  • Author(s): Takashi Kurashige, Hiroyuki Morino, Tomomi Murao, Yuishin Izumi, Tomohito Sugiura, Kazuya Kuraoka, Hideshi Kawakami, Tsuyoshi Torii, Hirofumi Maruyama
  • Organizer: 27th Ubterbatuibak Congress of the World Muscle Society
  • Int'l Joint Research
• [Presentation] ALSの栄養状態と予後の関連-CONUT scoreを用いた検討 (2022)
  • Author(s): 都甲めぐみ、中森正博、大下智彦、渡邉友彬、金谷雄平、内藤裕之、末田芳雅、北村健、野村栄一、鳥居剛、宮地隆史、山下拓史、渡邊千種、時信弘、丸山博文
  • Organizer: 第63回日本神経学会学術大会
• [Presentation] Natural history of amyotrophic lateral sclerosis with a Leu126Ser mutation in the SOD1 gene. (2021)
  • Author(s): T Matsubara, Y Izumi, T Komori, H Ito, M Oda, A Funai, K Hayashi, K Inoue, K Toyooka, M Kaido, H Fujimura, T Haraguchi, O Yokota, S Terada, Y Adachi, N Atsuta, G Sobue, M Aoki, H Morino, Hirofumi Maruyama, Y Saito, S Murayama
  • Organizer: The PanAsian Consortium for Treatment and Research in ALS
  • Int'l Joint Research
• [Presentation] Optineurin degects cause TDP43-pathology with autophagic vacuolar formation (2021)
  • Author(s): 倉重毅志、倉持真人、大澤亮介、山下結衣、塩井剛、森野豊之、鎌田正紀、綾木孝、伊東秀文、外丸祐介、丸山博文、川上秀史
  • Organizer: 第62回日本神経学会学術集会
• [Presentation] 孤発性筋萎縮性側索硬化症でのCFAP410発現の免疫組織学的検討 (2020)
  • Author(s): 倉重毅志、森野豊之、丸山博文、鳥居剛
  • Organizer: 第61回日本神経病理学会総会学術研究会
• [Presentation] Histone deacetylase 10 knockout activates chaperone-mediated autophagy and accelerates the decomposition of its substrate. (2020)
  • Author(s): Yoshito Nagano, Hitomi Obayashi, Tetsuya Takahashi, Takhiro Seki, Shigeru Tanaka, Norio Sakai, Masayasu Matsumoto, Hirofumi Maruyama
  • Organizer: 第61回日本神経学会学術大会
• [Book] 今日の治療指針 2021年版 筋萎縮性側索硬化症 (2021)
  • Author(s): 丸山博文
  • Total Pages: 2
  • Publisher: 医学書院
  • ISBN: 9784260042826
• [Book] 今日の診断指針第８版、筋萎縮 (2020)
  • Author(s): 丸山博文
  • Total Pages: 2
  • Publisher: 医学書院
  • ISBN: 9784260038096
• [Remarks] 筋萎縮性側索硬化症（ALS）診断の新規バイオマーカーの発見
  • URL: https://www.hiroshima-u.ac.jp/research/news/71002
• [Remarks] Muscle biopsy could lead to earlier diag of ALS
  • URL: https://www.sciencedaily.com/releases/2022/06/220621184506.htm