Analysis of molecular mechanisms regulating stemness in muscle stem cells

• Project/Area Number: 18K07082
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 49030:Experimental pathology-related
• Research Institution: National Center of Neurology and Psychiatry
• Principal Investigator: Hayashi Shinichiro 国立研究開発法人国立精神・神経医療研究センター, 神経研究所 疾病研究第一部, 室長 (10732381)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2020: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2019: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2018: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: 骨格筋 / 筋幹細胞 / 筋ジストロフィー / 筋衛星細胞 / 分化 / 未分化 / 幹細胞

Outline of Final Research Achievements

Satellite cells, a stem cell population in skeletal muscle, are essential for muscle regeneration. Transplanted satellite cells can fuse with endogenous muscle fibers and contribute regeneration in dystrophic muscle. However, once they are cultured in vitro, the ability of stemness maintenance is rapidly diminished. In this study, we aimed to understand molecular mechanisms to maintain stemness in satellite cells. We found that incubation of satellite cells with retinoic acid maintains satellite cell stemness and inhibits differentiation through retinoic acid receptor-alpha. In addition, laminin fragment also supports to inhibit satellite cell differentiation. This protocol improved engraftment of human satellite cells in dystrophic mice compared to conventional methods. Therefore, this new protocol may support ex vivo satellite cell expansion and stem cell therapy for treating dystrophy patients.

Academic Significance and Societal Importance of the Research Achievements

筋ジストロフィーの根治療法として骨格筋幹細胞である筋衛星細胞を移植する方法が挙げられるが、生体外で筋衛星細胞の未分化な状態を保持したまま培養することが難しく、移植に十分な数の筋衛星細胞を得られないことが課題となっている。本研究成果によって、これまでは困難であった骨格筋の幹細胞を生体外で未分化な状態のまま維持・増産できる事が可能となった。今後、レチノイン酸による筋分化抑制機構の分子メカニズムを明らかにすることによって細胞移植治療の発展に貢献できると期待される。

Research Products

• [Journal Article] Deficiency and overexpression of Rtl1 in the mouse cause distinct muscle abnormalities related to the Temple and Kagami-Ogata syndromes. (2020)
  • Author(s): Kitazawa M, Hayashi S, Imamura H, Takeda S, Oishi Y, Kaneko-Ishino T, Ishino F
  • Journal Title: Development Volume: 147 Issue: 21 Pages: 147-147
  • DOI: 10.1242/dev.185918
  • Peer Reviewed / Open Access
• [Journal Article] Pathologic Features of Anti-Mi-2 Dermatomyositis (2020)
  • Author(s): Tanboon Jantima、Inoue Michio、Hirakawa Shinya、Tachimori Hisateru、Hayashi Shinichiro、Noguchi Satoru、Suzuki Shigeaki、Okiyama Naoko、Fujimoto Manabu、Nishino Ichizo
  • Journal Title: Neurology Volume: 96 Issue: 3
  • DOI: 10.1212/wnl.0000000000011269
  • Peer Reviewed / Open Access
• [Journal Article] CGG expansion in NOTCH2NLC is associated with oculopharyngodistal myopathy with neurological manifestations (2020)
  • Author(s): Ogasawara M, Iida A, Kumutpongpanich T, Ozaki A, Oya Y, Konishi H, Nakamura A, Abe R, Takai H, Hanajima R, Doi H, Tanaka F, Nakamura H, Nonaka I, Wang Z, Hayashi S, Noguchi S, Nishino I.
  • Journal Title: Acta Neuropathol Commun Volume: 8 Issue: 1 Pages: 204-204
  • DOI: 10.1186/s40478-020-01084-4
  • Peer Reviewed / Open Access
• [Journal Article] ADSSL1 myopathy is the most common nemaline myopathy in Japan with variable clinical features (2020)
  • Author(s): Saito Yoshihiko、Nishikawa Atsuko、Iida Aritoshi、Mori-Yoshimura Madoka、Oya Yasushi、Ishiyama Akihiko、Komaki Hirofumi、Nakamura Seigo、Fujikawa Susumu、Kanda Takashi、Yamadera Misaki、Sakiyama Hiroshi、Hayashi Shinichiro、Nonaka Ikuya、Noguchi Satoru、Nishino Ichizo
  • Journal Title: Neurology Volume: 95 Issue: 11
  • DOI: 10.1212/wnl.0000000000010237
  • Peer Reviewed / Open Access
• [Journal Article] Two Japanese LGMDR25 patients with a biallelic recurrent nonsense variant of BVES (2020)
  • Author(s): Indrawati Luh Ari、Iida Aritoshi、Tanaka Yuzo、Honma Yutaka、Mizoguchi Koichi、Yamaguchi Tomohisa、Ikawa Masamichi、Hayashi Shinichiro、Noguchi Satoru、Nishino Ichizo
  • Journal Title: Neuromuscular Disorders Volume: 30 Issue: 8 Pages: 674-679
  • DOI: 10.1016/j.nmd.2020.06.004
  • Peer Reviewed / Open Access
• [Journal Article] Association of Dermatomyositis Sine Dermatitis With Anti-Nuclear Matrix Protein 2 Autoantibodies (2020)
  • Author(s): Michio Inoue, Kazumoto Shibuya (11th) et al.
  • Journal Title: JAMA Neurology Volume: 77 Issue: 7 Pages: 872-872
  • DOI: 10.1001/jamaneurol.2020.0673
  • Peer Reviewed / Open Access
• [Journal Article] Evaluation of the Core Formation Process in Congenital Neuromuscular Disease With Uniform Type 1 Fiber and Central Core Disease (2020)
  • Author(s): Ogasawara Masashi、Ogawa Megumu、Nonaka Ikuya、Hayashi Shinichiro、Noguchi Satoru、Nishino Ichizo
  • Journal Title: Journal of Neuropathology & Experimental Neurology Volume: 79 Issue: 12 Pages: 1370-1375
  • DOI: 10.1093/jnen/nlaa104
  • Peer Reviewed / Open Access
• [Journal Article] Exon skipping induced by nonsense/frameshift mutations in DMD gene results in Becker muscular dystrophy (2020)
  • Author(s): Okubo Mariko、Noguchi Satoru、Hayashi Shinichiro、Nakamura Harumasa、Komaki Hirofumi、Matsuo Masafumi、Nishino Ichizo
  • Journal Title: Human Genetics Volume: 139 Issue: 2 Pages: 247-255
  • DOI: 10.1007/s00439-019-02107-4
  • Peer Reviewed
• [Journal Article] PAX3 Confers Functional Heterogeneity in Skeletal Muscle Stem Cell Responses to Environmental Stress (2019)
  • Author(s): Der Vartanian Audrey、Qu?tin Marie、Michineau St?phanie、Aurad? Fr?d?ric、Hayashi Shinichiro、Dubois Christelle、Rocancourt Didier、Drayton-Libotte Bernadette、Szegedi Anik?、Buckingham Margaret、Conway Simon J.、Gervais Marianne、Relaix Fr?d?ric
  • Journal Title: Cell Stem Cell Volume: 24 Issue: 6 Pages: 958-973.e9
  • DOI: 10.1016/j.stem.2019.03.019
  • Peer Reviewed
• [Journal Article] COX6A2 Variants Cause a Muscle-Specific Cytochrome C Oxidase Deficiency (2019)
  • Author(s): Inoue Michio、Uchino Shumpei、Iida Aritoshi、Noguchi Satoru、Hayashi Shinichiro、Takahashi Tsutomu、Fujii Katsunori、Komaki Hirofumi、Takeshita Eri、Nonaka Ikuya、Okada Yukinori、Yoshizawa Takuya、Van Lommel Leentje、Schuit Frans、Goto Yu‐ichi、Mimaki Masakazu、Nishino Ichizo
  • Journal Title: Annals of Neurology Volume: 86 Issue: 2 Pages: 193-202
  • DOI: 10.1002/ana.25517
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Three novel MTM1 pathogenic variants identified in Japanese patients with X‐linked myotubular myopathy (2019)
  • Author(s): Nishikawa Atsuko、Iida Aritoshi、Hayashi Shinichiro、Okubo Mariko、Oya Yasushi、Yamanaka Gaku、Takahashi Ikuko、Nonaka Ikuya、Noguchi Satoru、Nishino Ichizo
  • Journal Title: Molecular Genetics & Genomic Medicine Volume: 7 Issue: 5
  • DOI: 10.1002/mgg3.621
  • Peer Reviewed
• [Journal Article] Three novel recessive DYSF mutations identified in three patients with muscular dystrophy, limb-girdle, type 2B. (2018)
  • Author(s): Okubo M, Iida A, Hayashi S, Mori-Yoshimura M, Oya Y, Watanabe A, Arahata H, El Sherif R, Noguchi S, Nishino I.
  • Journal Title: Journal of the Neurological Sciences Volume: 395 Pages: 169-171
  • DOI: 10.1016/j.jns.2018.10.015
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] A novel LMNA mutation identified in a Japanese patient with LMNA-associated congenital muscular dystrophy. (2018)
  • Author(s): Ishiyama A, Iida A, Hayashi S, Komaki H, Sasaki M, Nonaka I, Noguchi S, Nishino I.
  • Journal Title: Human Genome Variation Volume: 5 Issue: 1 Pages: 19-19
  • DOI: 10.1038/s41439-018-0018-6
  • Peer Reviewed / Open Access
• [Journal Article] Two novel VCP missense variants identified in Japanese patients with multisystem proteinopathy. (2018)
  • Author(s): Inoue M, Iida A, Hayashi S, Mori-Yoshimura M, Nagaoka A, Yoshimura S, Shiraishi H, Tsujino A, Takahashi Y, Nonaka I, Hayashi YK, Noguchi S, Nishino I.
  • Journal Title: Human Genome Variation Volume: 5 Issue: 1 Pages: 9-9
  • DOI: 10.1038/s41439-018-0009-7
  • Peer Reviewed / Open Access
• [Presentation] Myopathy in scleroderma/systemic sclerosis patients: From a histopathological point of view. (2020)
  • Author(s): Chen Y, Inoue M, Ogasawara M, Saito Y, Indrawati L, Tanboon J, Kumutpongpanich T, Okubo M, Yoshioka W, Hayashi S, Noguchi S, Nishino I
  • Organizer: 25th International Congress of the World Muscle Society
  • Int'l Joint Research
• [Presentation] Congenital neuromuscular disease with uniform type 1 fiber due to RYR1 mutation is a de facto core myopathy. (2020)
  • Author(s): Ogasawara M, Ogawa M, Nonaka I, Hayashi S, Noguchi S, Nishino I
  • Organizer: 25th International Congress of the World Muscle Society
  • Int'l Joint Research
• [Presentation] Generation and characterization of Pax7-HA knock-in mice. (2020)
  • Author(s): Hayashi S, Inoue Y, Kosako H, Inoue T, Noguchi S, Nishino I
  • Organizer: 25th International Congress of the World Muscle Society
  • Int'l Joint Research
• [Presentation] Screening for Pompe disease using muscle section in Japan. (2020)
  • Author(s): Saito Y, Nakamura K, Fukuda T, Sugie H, Hayashi S, Noguchi S, Nishino I
  • Organizer: 25th International Congress of the World Muscle Society
  • Int'l Joint Research
• [Presentation] A dominant variant in DNAJB4 causes a myofibrillar myopathy. (2020)
  • Author(s): Inoue M, Iida A, Watanabe K, Hosoi Y, Miyajima H, Hayashi S, Inoue Y, Inoue T, Noguchi S, Nishino I
  • Organizer: 25th International Congress of the World Muscle Society
  • Int'l Joint Research
• [Presentation] RYR1遺伝子異常によるCongenital neuromuscular disease with uniform type1 fiberは事実上のコアミオパチーである (2020)
  • Author(s): 小笠原真志，小川 恵，埜中征哉，林晋一郎，野口 悟，西野一三
  • Organizer: 第6回日本筋学会学術集会
• [Presentation] 本邦における骨格筋切片を用いたポンペ病スクリーニング (2020)
  • Author(s): 斎藤良彦，中村公俊，福田冬季子，杉江秀夫，林晋一郎，野口 悟，西野一三
  • Organizer: 第6回日本筋学会学術集会
• [Presentation] Recessive variant of POPDC3 causes a nuclear envelopathy. (2020)
  • Author(s): Indrawati LA, Mitsuhashi H, Iida A, Mori-Yoshimmura M, Hayashi S, Noguchi S, Nishino I
  • Organizer: 第6回日本筋学会学術集会
• [Presentation] RNA-seqを用いたジストロフィン遺伝子解析 (2020)
  • Author(s): 大久保真理子，野口 悟，林晋一郎，小牧宏文，齋藤伸治，西野一三
  • Organizer: 第6回日本筋学会学術集会
• [Presentation] Pathogenesis of congenital neuromuscular disease with uniform type 1 fiber and central core disease. (2020)
  • Author(s): Ogasawara M, Ogawa M, Hayashi S, Nonaka I, Noguchi S, Nishino I
  • Organizer: 第62回日本小児神経学会学術集会
• [Presentation] 筋発生・再生過程における骨格筋細胞の分化制御機構 (2019)
  • Author(s): 林 晋一郎
  • Organizer: 第74回日本体力医学会大会
  • Invited
• [Presentation] 骨格筋の再生・発生過程における筋細胞の分化制御メカニズム (2019)
  • Author(s): 林 晋一郎
  • Organizer: 第92回日本生化学会大会
  • Invited
• [Presentation] Genetic diagnosis in large Japanese cohort using targeted re-sequencing system (2019)
  • Author(s): Okubo M, Iida A, Inoue M, Saito Y, Ogasawara M, Hayashi S, Noguchi S, Nishino I
  • Organizer: 24th International Congress of the World Muscle Society
  • Int'l Joint Research
• [Presentation] ADSSL1 myopathy is a fatigability disease presenting both nemaline bodies and lipid droplets in skeletal muscles-a study of 57 Japanese cases (2019)
  • Author(s): Saito Y, Noguchi S, Nishikawa A, Hayashi S, Iida A, Nishino I
  • Organizer: 24th International Congress of the World Muscle Society
  • Int'l Joint Research
• [Presentation] ETS1 facilitates muscle satellite cell proliferation and stemness perpetuation (2019)
  • Author(s): Kumutpongpanich T, Hayashi S, Noguchi S, Nishino I
  • Organizer: 24th International Congress of the World Muscle Society
  • Int'l Joint Research
• [Presentation] Two murine models for tubular aggregate myopathy with mutations in Stim1 and Orai1 (2019)
  • Author(s): Ogasawara M, Ogawa M, Endo Y, Lee J, Inoue Y, Inoue T, Nishino I, Noguchi S
  • Organizer: 24th International Congress of the World Muscle Society
  • Int'l Joint Research
• [Presentation] Mutation-specific therapy for X-linked myotubular myopathy (2019)
  • Author(s): Hayashi S, Noguchi S, Kumutpongpanich T, Iida A, Okubo M, Matsuo M, Nishino I
  • Organizer: 24th International Congress of the World Muscle Society
  • Int'l Joint Research
• [Presentation] Infantile-onset lipid storage myopathy. (2019)
  • Author(s): Indrawati LA, Noguchi S, Tanboon J, Ogasawara M, Saito Y, Kumutpongpanich T, Inoue M, Okubo M, Fukuda T, Sugie H, Goto Y, Iida A, Hayashi S, Nishino I
  • Organizer: 24th International Congress of the World Muscle Society
  • Int'l Joint Research
• [Presentation] 次世代シークエンサーを用いたデュシェンヌ/ベッカー型筋ジストロフィーの遺伝学的解析（続報） (2019)
  • Author(s): 大久保真理子，野口 悟，飯田有俊，林 晋一郎，西野一三
  • Organizer: 第64回日本人類遺伝学会
• [Presentation] COX6A2変異は横紋筋特異的なチトクロームc酸化酵素欠損症を引き起こす (2019)
  • Author(s): 井上道雄，内野俊平，飯田有俊，野口 悟，林 晋一郎，高橋 努，藤井克則，小牧宏文，竹下絵里，埜中征哉，岡田随象，吉澤拓也，Van Lommel L, Schuit F, 後藤雄一，三牧正和，西野一三
  • Organizer: 日本筋学会第5回学術集会
• [Presentation] GNEミオパチーp.D207V変異は軽症型変異であり、ホモ接合体では極めて稀にしか発症しない (2019)
  • Author(s): 吉岡和香子，飯田有俊，岡田随象，大矢 寧，大久保真理子，小川 恵，松田文彦，日笠幸一郎，森 まどか，中村治雅，林 晋一郎，野口 悟，西野一三
  • Organizer: 日本筋学会第5回学術集会
• [Presentation] ETS1 regulates muscle stem cell proliferation and stemness perpetuation (2019)
  • Author(s): Kumutpongpanich T, Hayashi S, Noguchi S, Nishino I
  • Organizer: 日本筋学会第5回学術集会
• [Presentation] Establishment of a long-term culture system for Muscle stem cells. (2018)
  • Author(s): Hayashi S.
  • Organizer: 5th Tissue Engineering and Regenerative Medicine International Society World Congress
  • Int'l Joint Research / Invited
• [Presentation] Klf5 has essential roles in myoblast differentiation and survival during fetal muscle development. (2018)
  • Author(s): Hayashi S, Oishi Y, Manabe I, Nishino I.
  • Organizer: 23rd INTERNATIONAL CONGRESS OF THE WORLD MUSCLE SOCIETY
  • Int'l Joint Research
• [Presentation] Nonsense mutation induced exon skipping in Becker muscular dystrophy. (2018)
  • Author(s): Okubo M, Noguchi S, Hayashi S, Matsuo M, Nishino I.
  • Organizer: 23rd INTERNATIONAL CONGRESS OF THE WORLD MUSCLE SOCIETY
  • Int'l Joint Research
• [Presentation] Novel deep intronic mutations in MTM1 in X-linked myotubular myopathy. (2018)
  • Author(s): Kumutpongpanich T, Hayashi S, Iida A, Okubo M, Matsuo M, Noguchi S, Nishino I
  • Organizer: 23rd INTERNATIONAL CONGRESS OF THE WORLD MUSCLE SOCIETY
  • Int'l Joint Research
• [Presentation] Regulation of muscle stem cell function during murine fetal development. (2018)
  • Author(s): Hayashi S, Manabe I, Oishi Y, Nishino I.
  • Organizer: 11th Japanese-French Workshop “New insights in personalized medicine for neuromuscular diseases: From Basic to Applied Myology”
  • Int'l Joint Research
• [Presentation] Frequency of c.620A>T homozygotes is extremely low in Japanese GNE myopathy patients: What is determining the pathogenicity? (2018)
  • Author(s): Yoshioka W, Okubo M, Ogawa M, Matsuda F, Higasa K, Mori-Yoshimura M, Oya Y, Kimura E, Nakamura H, Hayashi S, Iida A, Noguchi S, Nishino I
  • Organizer: 11th Japanese-French Workshop “New insights in personalized medicine for neuromuscular diseases: From Basic to Applied Myology”
  • Int'l Joint Research
• [Presentation] Comprehensive analysis in Japan: Nonsense mutation induced exon skipping in Becker muscular dystrophy. (2018)
  • Author(s): Okubo M, Noguchi S, Hayashi S, Nishino I.
  • Organizer: 11th Japanese-French Workshop “New insights in personalized medicine for neuromuscular diseases: From Basic to Applied Myology”
  • Int'l Joint Research
• [Presentation] Klf5 regulates myoblast differentiation and apoptosis during fetal muscle development. (2018)
  • Author(s): Hayashi S.
  • Organizer: 第6回若手による骨格筋細胞研究会
• [Presentation] Klf5による骨格筋発生の制御機構 (2018)
  • Author(s): 林晋一郎，大石由美子，真鍋一郎，西野一三
  • Organizer: 日本筋学会第4回学術集会
• [Book] 超高齢社会に挑む骨格筋のメディカルサイエンス (2018)
  • Author(s): 武田 伸一, 林晋一郎, 他
  • Total Pages: 230
  • Publisher: 羊土社
  • ISBN: 9784758103701