Elucidation of molecular mechanisms of neuromuscular junction formations in physiology and pathology

2020 Fiscal Year Annual Research Report

• Project/Area Number: 20H03561
• Research Institution: Nagoya University
• Principal Investigator: 大野 欽司 名古屋大学, 医学系研究科, 教授 (80397455)
• Co-Investigator(Kenkyū-buntansha): 増田 章男 名古屋大学, 医学系研究科, 准教授 (10343203) ; 大河原 美静 名古屋大学, 医学系研究科, 准教授 (80589606)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Keywords: 神経筋接合部 / 先天性筋無力症候群

Outline of Annual Research Achievements

神経筋接合部(neuromuscular junction, NMJ)の正常分子構築機構を明らかにするとともに、先天性筋無力症候群(congenital myasthenic syndromes, CMS)の分子病態を解明し、新規治療法開発につなげることを目的とした。CMSはNMJ発現分子の先天的遺伝子欠損によりNMJ信号伝達が障害をされる病態であり、出生時を含む２歳以下発症が多く、筋力低下と日内変動と日差変動を特徴とし、時に顔面の小奇形を伴う。今までに30種類以上の原因遺伝子が同定されてきた。本研究では、初めに、Rspo2, Fgf18, Ctgfの3種類の新規NMJ構築誘導分子とその受容体の機能解析により正常NMJ分子構築機構を明らかにするとともに、RiboTagマウスとSPPLAT法によるproximity proteomicsにより新規NMJ構築分子同定を行った。次に、本邦CMSにおいて同定を行なってきた遺伝子変異の機能解析を培養細胞を用いて行うことを目的としてin vitro NMJの作成を開始した。これらのCMS病態解析を新規治療法開発研究に今後つなげる。

Current Status of Research Progress

2: Research has progressed on the whole more than it was originally planned.

Reason

COVID-19パンデミックによる研究の一部遅延が生じているが概ね順調に経過している。

Strategy for Future Research Activity

CMSモデルマウスとRiboTagマウスをかけあわせることによりCMSのNMJ筋終板ならびに脊髄前角細胞における遺伝子発現変化を解析しCMS病態を明らかにする。iPS細胞を用いたin vitro NMJを作成し、CMS病態を明らかにするとともに、薬剤スクリーニングで同定をした新規治療薬の効果の検証を行う。

Research Products

• [Journal Article] Efficacy of salbutamol monotherapy in slow‐channel congenital myasthenic syndrome caused by a novel mutation in CHRND (2021)
  • Author(s): Tawara Nozomu、Yamashita Satoshi、Takamatsu Koutaro、Yamasaki Yoshimune、Mukaino Akihiro、Nakane Shunya、Farshadyeganeh Paniz、Ohno Kinji、Ando Yukio
  • Journal Title: Muscle & Nerve Volume: 63 Pages: E30-E32
  • DOI: 10.1002/mus.27166
  • Peer Reviewed
• [Journal Article] Secreted Signaling Molecules at the Neuromuscular Junction in Physiology and Pathology (2021)
  • Author(s): Ohkawara Bisei、Ito Mikako、Ohno Kinji
  • Journal Title: International Journal of Molecular Sciences Volume: 22 Pages: 2455～2455
  • DOI: 10.3390/ijms22052455
  • Peer Reviewed / Open Access
• [Journal Article] Screening of signaling pathways for agrin-induced AChR clustering in C2C12 myotubes (2021)
  • Author(s): Noto K, Ohkawara B, Ishii H, Ito M, Masuda A, Hirata H, Ohno K.
  • Journal Title: Genes Cells Volume: - Pages: in press.
  • Peer Reviewed
• [Journal Article] Hierarchical non-negative matrix factorization using clinical information for microbial communities (2021)
  • Author(s): Abe Ko、Hirayama Masaaki、Ohno Kinji、Shimamura Teppei
  • Journal Title: BMC Genomics Volume: 22 Pages: 104-104
  • DOI: 10.1186/s12864-021-07401-y
  • Peer Reviewed / Open Access
• [Journal Article] tRIP‐seq reveals repression of premature polyadenylation by co‐transcriptional FUS‐U1 snRNP assembly (2020)
  • Author(s): Masuda Akio、Kawachi Toshihiko、Takeda Jun‐ichi、Ohkawara Bisei、Ito Mikako、Ohno Kinji
  • Journal Title: EMBO reports Volume: 21 Pages: e49890
  • DOI: 10.15252/embr.201949890
  • Peer Reviewed / Open Access
• [Journal Article] Congenital myasthenic syndrome?associated agrin variants affect clustering of acetylcholine receptors in a domain-specific manner (2020)
  • Author(s): Ohkawara Bisei、Shen XinMing、Selcen Duygu、Nazim Mohammad、Bril Vera、Tarnopolsky Mark A.、Brady Lauren、Fukami Sae、Amato Anthony A.、Yis Uluc、Ohno Kinji、Engel Andrew G.
  • Journal Title: JCI Insight Volume: 5 Pages: e132023
  • DOI: 10.1172/jci.insight.132023
  • Peer Reviewed / Open Access
• [Journal Article] Meta‐Analysis of Gut Dysbiosis in Parkinson's Disease (2020)
  • Author(s): Nishiwaki Hiroshi、Ito Mikako、Ishida Tomohiro、Hamaguchi Tomonari、Maeda Tetsuya、Kashihara Kenichi、Tsuboi Yoshio、Ueyama Jun、Shimamura Teppei、Mori Hiroshi、Kurokawa Ken、Katsuno Masahisa、Hirayama Masaaki、Ohno Kinji
  • Journal Title: Movement Disorders Volume: 35 Pages: 1626～1635
  • DOI: 10.1002/mds.28119
  • Peer Reviewed / Open Access
• [Journal Article] InMeRF: prediction of pathogenicity of missense variants by individual modeling for each amino acid substitution (2020)
  • Author(s): Takeda Jun-ichi、Nanatsue Kentaro、Yamagishi Ryosuke、Ito Mikako、Haga Nobuhiko、Hirata Hiromi、Ogi Tomoo、Ohno Kinji
  • Journal Title: NAR Genomics and Bioinformatics Volume: 2 Pages: lqaa038
  • DOI: 10.1093/nargab/lqaa038
  • Peer Reviewed
• [Journal Article] CTGF/CCN2 facilitates LRP4‐mediated formation of the embryonic neuromuscular junction (2020)
  • Author(s): Ohkawara Bisei、Kobayakawa Akinori、Kanbara Shunsuke、Hattori Takako、Kubota Satoshi、Ito Mikako、Masuda Akio、Takigawa Masaharu、Lyons Karen M、Ishiguro Naoki、Ohno Kinji
  • Journal Title: EMBO reports Volume: 21 Pages: e48462
  • DOI: 10.15252/embr.201948462
  • Peer Reviewed / Open Access
• [Journal Article] Zonisamide ameliorates progression of cervical spondylotic myelopathy in a rat model (2020)
  • Author(s): Kanbara Shunsuke、Ohkawara Bisei、Nakashima Hiroaki、Ohta Kyotaro、Koshimizu Hiroyuki、Inoue Taro、Tomita Hiroyuki、Ito Mikako、Masuda Akio、Ishiguro Naoki、Imagama Shiro、Ohno Kinji
  • Journal Title: Scientific Reports Volume: 10 Pages: 13138-13138
  • DOI: 10.1038/s41598-020-70068-0
  • Peer Reviewed / Open Access
• [Journal Article] Gene Expression Profile at the Motor Endplate of the Neuromuscular Junction of Fast-Twitch Muscle (2020)
  • Author(s): Huang Kun、Li Jin、Ito Mikako、Takeda Jun-Ichi、Ohkawara Bisei、Ogi Tomoo、Masuda Akio、Ohno Kinji
  • Journal Title: Frontiers in Molecular Neuroscience Volume: 13 Pages: 154-154
  • DOI: 10.3389/fnmol.2020.00154
  • Peer Reviewed / Open Access
• [Journal Article] Short-Chain Fatty Acid-Producing Gut Microbiota Is Decreased in Parkinson’s Disease but Not in Rapid-Eye-Movement Sleep Behavior Disorder (2020)
  • Author(s): Nishiwaki Hiroshi、Hamaguchi Tomonari、Ito Mikako、Ishida Tomohiro、Maeda Tetsuya、Kashihara Kenichi、Tsuboi Yoshio、Ueyama Jun、Shimamura Teppei、Mori Hiroshi、Kurokawa Ken、Katsuno Masahisa、Hirayama Masaaki、Ohno Kinji
  • Journal Title: mSystems Volume: 5 Pages: e00797-20
  • DOI: 10.1128/mSystems.00797-20
  • Peer Reviewed / Open Access
• [Journal Article] Identification of Qk as a Glial Precursor Cell Marker that Governs the Fate Specification of Neural Stem Cells to a Glial Cell Lineage (2020)
  • Author(s): Takeuchi Akihide、Takahashi Yuji、Iida Kei、Hosokawa Motoyasu、Irie Koichiro、Ito Mikako、Brown J.B.、Ohno Kinji、Nakashima Kinichi、Hagiwara Masatoshi
  • Journal Title: Stem Cell Reports Volume: 15 Pages: 883～897
  • DOI: 10.1016/j.stemcr.2020.08.010
  • Peer Reviewed / Open Access
• [Journal Article] Zonisamide ameliorates neuropathic pain partly by suppressing microglial activation in the spinal cord in a mouse model (2020)
  • Author(s): Koshimizu Hiroyuki、Ohkawara Bisei、Nakashima Hiroaki、Ota Kyotaro、Kanbara Shunsuke、Inoue Taro、Tomita Hiroyuki、Sayo Akira、Kiryu-Seo Sumiko、Konishi Hiroyuki、Ito Mikako、Masuda Akio、Ishiguro Naoki、Imagama Shiro、Kiyama Hiroshi、Ohno Kinji
  • Journal Title: Life Sciences Volume: 263 Pages: 118577～118577
  • DOI: 10.1016/j.lfs.2020.118577
  • Peer Reviewed
• [Presentation] InMeRF: A set of random forest models for each amino acid substitution to predict pathogenicity of missense variants in the human genome (2020)
  • Author(s): Takeda J, Nanatsue K, Yamagishi R, Ito M, Ohno K
  • Organizer: European Society of Human Genetics Virtual Conference
  • Int'l Joint Research