Elucidation of the molecular pathogenesis of selective degeneration in multisystem proteinopathy and amyotrophic lateral sclerosis.

• Project/Area Number: 20K07897
• 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: Tohoku University
• Principal Investigator: Warita Hitoshi 東北大学, 大学病院, 助教 (30400245)
• 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: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2021: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2020: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: 多系統蛋白質症 / 筋萎縮性側索硬化症 / 骨格筋 / RNA結合蛋白 / hnRNP / 運動ニューロン / オミックス解析 / トランスクリプトーム

Outline of Research at the Start

本研究では、HNRNPA1変異に伴う多系統蛋白質症（MSP）-筋萎縮性側索硬化症（ALS）で選択的変性に陥る細胞群（大脳皮質ニューロン，脊髄運動ニューロン，骨格筋細胞）と変性を免れやすい細胞群（外眼筋支配運動ニューロンや感覚ニューロン）を樹立済の自験MSP3および対照ヒトiPS細胞から分化誘導し、それらの比較研究から(1) 単純・共培養系＋ストレス負荷における細胞表現型と分子細胞病理，各細胞間の相違，(2) 各細胞群の全トランスクリプトーム比較解析で抽出される病態関連分子とその相違，(3) 各細胞群においてhnRNPA1と相互作用するRNA種を明らかにして、治療標的分子の発見をめざす。

Outline of Final Research Achievements

To elucidate the cell-selective degeneration mechanism in multisystem proteinopathy (MSP) and amyotrophic lateral sclerosis (ALS), we established multiple lines of MSP3 patients-derived pluripotent stem cells (iPSCs). Motor neurons and skeletal muscle cells defferentiated from the MSP3-specific iPSCs and isogenic control cells were subjected to this study. After improving the efficiency of induction and differentiation into skeletal muscle cells and the long-term culture method, a comparative analyses was performed to reveal molecular cell pathology. As a result, we found that hnRNPA1 was localized to the cytoplasm under oxidative stress as a pathological phenotype of skeletal muscle cells.

Academic Significance and Societal Importance of the Research Achievements

本研究成果をふまえ、スペクトラムを形成しつつある多系統蛋白質症（MSP）－筋萎縮性側索硬化症（ALS）の原因遺伝子変異がもたらす細胞種選択的な変性メカニズムの解明と治療標的分子の発見につなげることで、治療法が未確立のこれら神経変性疾患のみならず、他の運動ニューロン疾患や骨格筋変性疾患の治療法開発にもつながる意義がある。

Research Products

• [Journal Article] MOG Antibody-Associated Disorders Following SARS- CoV-2 Vaccination: A Case Report and Literature Review. (2022)
  • Author(s): Matsumoto Y, Ohyama A, Kubota T, Ikeda K, Kaneko K, Takai Y, Warita H, Takahashi T, Misu T, Aoki M.
  • Journal Title: Front Neurol. Volume: 13 Pages: 845755-845755
  • DOI: 10.3389/fneur.2022.845755
  • Peer Reviewed / Open Access
• [Journal Article] Long-term outcomes after surgery to prevent aspiration for patients with amyotrophic lateral sclerosis (2022)
  • Author(s): Soga T, Suzuki N, Kato K, Kawamoto-Hirano A, Kawauchi Y, Izumi R, Toyoshima M, Mitsuzawa S, Shijo T, Ikeda K, Warita H, Katori Y, Aoki M, Kato M.
  • Journal Title: BMC Neurology Volume: 22 Issue: 1 Pages: 94-94
  • DOI: 10.1186/s12883-022-02619-z
  • Peer Reviewed / Open Access
• [Journal Article] Malignant otitis externa presenting cerebral infarction from pseudoaneurysm: A case report and a review of the literature (2022)
  • Author(s): Koshiba Y, Ikeda R, Suzuki J, Honkura Y, Funayama Y, Ikeda K, Warita H, Aoki M, Kawase T, Katori Y.
  • Journal Title: Clinical Case Reports Volume: 10 Issue: 2
  • DOI: 10.1002/ccr3.5276
  • Peer Reviewed / Open Access
• [Journal Article] A novel deletion in the C-terminal region of HSPB8 in a family with rimmed vacuolar myopathy (2021)
  • Author(s): Inoue-Shibui Aya、Niihori Tetsuya、Kobayashi Michio、Suzuki Naoki、Izumi Rumiko、Warita Hitoshi、Hara Kenju、Shirota Matsuyuki、Funayama Ryo、Nakayama Keiko、Nishino Ichizo、Aoki Masashi、Aoki Yoko
  • Journal Title: Journal of Human Genetics Volume: NA Issue: 10 Pages: 965-972
  • DOI: 10.1038/s10038-021-00916-y
  • Peer Reviewed
• [Journal Article] Reduced PHOX2B stability causes axonal growth impairment in motor neurons with TARDBP mutations (2021)
  • Author(s): Mitsuzawa S, Suzuki N, Akiyama T, Ishikawa M, Sone T, Kawada J, Funayama R, Shirota M, Mitsuhashi H, Morimoto S, Ikeda K, Shijo T, Ohno A, Nakamura N, Ono H, Ono R, Osana S, Nakagawa T, Nishiyama A, Izumi R, Kaneda S, Ikeuchi Y, Nakayama K, Fujii T, Warita H, Okano H, Aoki M.
  • Journal Title: Stem Cell Reports Volume: 16 Issue: 6 Pages: 1527-1541
  • DOI: 10.1016/j.stemcr.2021.04.021
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Randomized phase 2 study of perampanel for sporadic amyotrophic lateral sclerosis (2021)
  • Author(s): Aizawa H, Kato H, Oba K, Kawahara T, Okubo Y, Saito T, Naito M, Urushitani M, Tamaoka A, Nakamagoe K, Ishii K, Kanda T, Katsuno M, Atsuta N, Maeda Y, Nagai M, Nishiyama K, Ishiura H, Toda T, Kawata A, Abe K, Yabe I, Takahashi-Iwata I, Sasaki H, Warita H, Aoki M, Sobue G, Mizusawa H, Matsuyama Y, Haga T, Kwak S
  • Journal Title: Journal of Neurology Volume: 269 Issue: 2 Pages: 885-896
  • DOI: 10.1007/s00415-021-10670-y
  • Peer Reviewed / Open Access
• [Journal Article] RNP顆粒生成・分解からみた多系統蛋白質症 (2021)
  • Author(s): 割田仁, 池田謙輔, 青木正志
  • Journal Title: 脳神経内科 Volume: 95 Pages: 112-120
• [Journal Article] Generation of an ALS human iPSC line KEIOi001-A from peripheral blood of a Charcot disease-affected patient carrying TARDBP p.N345K heterozygous SNP mutation (2020)
  • Author(s): Leventoux Nicolas、Morimoto Satoru、Hara Kenju、Nakamura Shiho、Ozawa Fumiko、Mitsuzawa Shio、Akiyama Tetsuya、Nishiyama Ayumi、Suzuki Naoki、Warita Hitoshi、Aoki Masashi、Okano Hideyuki
  • Journal Title: Stem Cell Research Volume: 47 Pages: 101896-101896
  • DOI: 10.1016/j.scr.2020.101896
  • Peer Reviewed / Open Access
• [Journal Article] An Amyotrophic Lateral Sclerosis-Associated Mutant of C21ORF2 Is Stabilized by NEK1-Mediated Hyperphosphorylation and the Inability to Bind FBXO3 (2020)
  • Author(s): Watanabe Y, Nakagawa T, Akiyama T, Nakagawa M, Suzuki N, Warita H, Aoki M, Nakayama K.
  • Journal Title: iScience Volume: 23 Issue: 9 Pages: 1-15
  • DOI: 10.1016/j.isci.2020.101491
  • Peer Reviewed / Open Access
• [Presentation] Motor nerve organoid is useful tool to analyze axonal degeneration of ALS. (2021)
  • Author(s): Suzuki N, Kawada J, Akiyama T, Mitsuzawa S, Kato M, Warita H, Fujii T, Eggan K, Okano H, Aoki M.
  • Organizer: 第62回日本神経学会学術大会（62st Annual Meeting of the Japanese Society of Neurology）
• [Presentation] Axonal growth impairment in iPS-derived motor neurons with TARDBP mutations. (2021)
  • Author(s): Mitsuzawa S, Suzuki N, Akiyama T, Ishikawa M, Sone T, Kawada J, Funayama R, Mitsuhashi H, Nishiyama A, Ikeda K, Shijo T, Nakamura N, Ono Y, Ono R, Izumi R, Ohno A, Nakagawa T, Nakayama K, Warita H, Okano H, Aoki M.
  • Organizer: 第62回日本神経学会学術大会（62st Annual Meeting of the Japanese Society of Neurology）
• [Presentation] Reduced PHOX2B stability causes axonal growth impairment in motor neurons with TARDBP mutations. (2021)
  • Author(s): Mitsuzawa S, Suzuki N, Akiyama T, Ishikawa M, Sone T, Kawada J, Mitsuhashi H, Morimoto S, Ikeda K, Shijo T, Ohno A, Nakamura N, Ono H, Ono R, Nishiyama A, Izumi R, Kato M, Warita H, Okano H, Aoki M.
  • Organizer: PACTALS 2021
  • Int'l Joint Research
• [Presentation] Reduced PHOX2B stability causes axonal growth impairment in motor neurons with ALS-linked TARDBP mutations. (2021)
  • Author(s): Mitsuzawa S, Suzuki N, Akiyama T, Ishikawa M, Sone T, Kawada J, Mitsuhashi H, Morimoto S, Ikeda K, Shijo T, Ohno A, Nakamura N, Ono H, Ono R, Nishiyama A, Izumi R, Kaneda S, Ikeuchi Y, Nakayama K, Fujii T, Warita H, Okano H, Aoki M.
  • Organizer: ISSCR 2021 Tokyo
  • Int'l Joint Research
• [Presentation] Axonal growth impairment in motor neurons with TARDBP mutations were mediated by PHOX2B downregulation. (2021)
  • Author(s): Mitsuzawa S, Suzuki N, Akiyama T, Ishikawa M, Sone T, Kawada J, Funayama R, Shirota M, Mitsuhashi H, Morimoto S, Ikeda K, Shijo T, Ohno A, Nakamura N, Ono H, Ono R, Nishiyama A, Izumi R, Kaneda S, Ikeuchi Y, Nakayama K, Fujii T, Kato M, Warita H, Okano H, Aoki M.
  • Organizer: 32th International Symposium on ALS/MND
  • Int'l Joint Research
• [Presentation] ALSに対する肝細胞増殖因子（HGF）脊髄腔内反復投与の第II相試験 (2020)
  • Author(s): 割田 仁
  • Organizer: 第61回日本神経学会学術大会
  • Invited
• [Book] 脳神経疾患最新の治療2021-2023 (2020)
  • Author(s): 割田 仁，青木正志（分担執筆，園生雅弘・北川一夫・青木正志 共同編集）
  • Total Pages: 368
  • Publisher: 南江堂
  • ISBN: 4524227857
• [Remarks] 東北大学神経内科
  • URL: http://www.neurol.med.tohoku.ac.jp/index.html
• [Remarks] Researchmap
  • URL: https://researchmap.jp/warita_2012
• [Remarks] ResearchGate
  • URL: https://www.researchgate.net/profile/Hitoshi-Warita
• [Remarks] ORCiD
  • URL: https://orcid.org/0000-0001-7934-3863