神経筋接合分子欠損症におけるmRNA病態研究と治療法開発研究

2011 Fiscal Year Annual Research Report

• Project/Area Number: 21390266
• 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, 助教 (10343203) ; 伊藤 美佳子 名古屋大学, 医学系研究科, 助教 (60444402)
• Keywords: 先天性筋無力症候群 / 終板アセチルコリンエステラーゼ欠損症 / protein-anchoring therapy / Collagen Q

Research Abstract

先天性筋無力症候群に対するタンパク標的治療法(protein-anchoring therapy)の開発を行った。先天性筋無力症候群の一型である終板アセチルコリンエステラーゼ(AChE)欠損症はAChEをシナプス基底膜に係留するcollagen Q(ColQ)分子の先天的な欠損によっておきる。本症に対する特異的な治療法は存在しない。研究代表者らはヒトCOLQをAAVベクターに組み込み、骨格筋と肝臓に親和性の高いserotype 8 capsidを用いて作成をしたAAV8-COLQウィルス粒子をColqノックアウトマウスに尾静脈より投与した。電気生理学的検討にてMEPP減衰時間の正常化、反復神経刺激におけるCMAP減衰の改善、筋終板電位の正常化を認め、組織化学的検討にて観察をしたすべての神経筋接合部においてAchE/ColQ複合体の発現を認めた。さらに生化学的検討にて骨格筋AChE/ColQ複合体は正常の89%まで回復をしており、その分子構成も正常化していた。さらに運動機能も正常マウスと同等まで治療後3週で回復し、最高1年半効果が持続した。次に、全身への移行が少ないAAVl-COLQを作成し一側下肢に筋注を行いAChE/ColQ複合体の四肢筋への分布を確認した。さらに精製AchE/ColQ複合体を大量に作成しColq-/-マウス臀筋に持続的に筋肉注射を行いAChE/ColQ複合体の前肢を含む全身骨格筋への分布を確認した。タンパク分子自身が標的組織への親和性を持つ細胞外分子の特徴を生かしたprotein-anchoring therapyの有用性を終板AChE欠損症モデルマウスで実証し他の細胞外分子欠損症に応用が可能である可能性を示した。

Research Products

• [Journal Article] A novel mutation in SCN4A causes severe myotonia and school-age-onset paralytic episodes (2012)
  • Author(s): Yoshinaga H, Sakoda S, Good JM, Takahashi MP, Kubota T, Arikawa-Hirasawa E, Nakata T, OhnoK, Kitamura T, Kobayashi K, Ohtsuka Y
  • Journal Title: J Neurol Sci Volume: 315 Pages: 15-19
  • Peer Reviewed
• [Journal Article] CUGBP1 and MBNL1 preferentially bind to 3' UTRs and facilitate mRNA decay (2012)
  • Author(s): Masuda A, Andersen HS, Doktor TK, Okamoto T, Ito M, Andresen BS, Ohno K
  • Journal Title: Scientific Reports Volume: 2 Pages: 209
  • Peer Reviewed
• [Journal Article] Myotonic dystrophy type 2 (DM2) is rare in the Japanese population (2012)
  • Author(s): Matsuura T, Minami N, Arahata H, Ohno K, Abe K, Hayashi YK, Nishino I
  • Journal Title: J Hum Genet Volume: (印刷中)
  • DOI: 10.1038/jhg.2011.152 jhg2011152[pii]
  • Peer Reviewed
• [Journal Article] Protein-anchoring strategy for delivering acetylcholinesterase to the neuromuscular junction (2012)
  • Author(s): Ito M, Suzuki Y, Okada T, Fukudome T, Yoshimura T, Masuda A, Takeda S, Krejci E, Ohno K
  • Journal Title: Mol Ther Volume: (印刷中)
  • DOI: 10.1038/mt.2012.34 mt201234[pii]
  • Peer Reviewed
• [Journal Article] Anti-MuSK autoantibodies block binding of collagen Q to MuSK (2011)
  • Author(s): Kawakami Y, Ito M, Hirayama M, Sahashi K, Ohkawara B, Masuda A, Nishida H, Mabuchi N, Engel A G, Ohno K
  • Journal Title: Neurology Volume: 77 Pages: 1819-1826
  • Peer Reviewed
• [Journal Article] Myasthenic syndrome caused by plectinopathy (2011)
  • Author(s): Selcen D, Juel VC, Hobson-Webb LD, Smith EC, Stickler DE, Bite AV, Ohno K, Engel AG
  • Journal Title: Neurology Volume: 76 Pages: 327-336
  • Peer Reviewed
• [Journal Article] AG-dependent 3'-splice sites are predisposed to aberrant splicing due to a mutation at the first nucleotide of an exon (2011)
  • Author(s): Fu Y, Masuda A, Ito M, Shinmi J, Ohno K
  • Journal Title: Nucleic Acids Research Volume: 39 Pages: 4396-4404
  • DOI: gkr026[pii] 10.1093/nar/gkr026
  • Peer Reviewed
• [Journal Article] Molecular hydrogen inhibits lipopolysaccharide/interferon gamma-induced nitric oxide production through modulation of signal transduction in macrophages (2011)
  • Author(s): Itoh T, Hamada N, Terazawa R, Ito M, Ohno K, Ichihara M, Nozawa Y, Ito M
  • Journal Title: Biochem Biophys Res Commun Volume: 411 Pages: 143-149
  • DOI: S0006-291X(11)01096-5[pii] 10.1016/j.bbrc.2011.06.116
  • Peer Reviewed
• [Journal Article] Hyperuricemia cosegregating with osteogenesis imperfecta is associated with a mutation in GPATCH8 (2011)
  • Author(s): Kaneko H, Kitoh H, Matsuura T, Masuda A, Ito M, Mottes M, Rauch F, Ishiguro N, Ohno K
  • Journal Title: Hum Genet Volume: 130 Pages: 671-683
  • DOI: 10.1007/s00439-011-1006-9
  • Peer Reviewed
• [Presentation] Anti-MuSK antibodies in myasthenia gravis block binding of collagen Q to MuSK expressed at the neuromuscular junction (2011)
  • Author(s): Kawakami Y, Ito M, Hirayama M, Sahashi K, Ohkawara B, Masuda A, Hishida H, Mabuchi N, Engel AG, Ohno K
  • Organizer: 41st Annual Meeting, Society for Neuroscience
  • Place of Presentation: Washington DC, USA
  • Year and Date: 2011-11-15
• [Presentation] Protein-anchoring therapy for delivering acetylcholinesterase to the neuromuscular junction (2011)
  • Author(s): Ito M, Suzuki Y, Okada T, Fukudome T, Yoshimura T, Masuda A, Takeda S, Krejci E, Ohno K
  • Organizer: 4th International Congress of Myology
  • Place of Presentation: Lille, France(招待講演)
  • Year and Date: 2011-05-09
• [Book] Congenital myasthenic syndromes. Myasthenia gravis and myasthenic disorders 2nd ed.(Ed.by Engel AG) (2012)
  • Author(s): Engel AG, Shen X-M, Ohno K, Sine SM
  • Publisher: Oxford University Press, New York(in press)
• [Book] Congenital Myasthenic Syndromes-Molecular Bases of Congenital Defects of Proteins at the Neuromuscular Junction-Myopathy (2012)
  • Author(s): Ohno K, Ito M, Engel AG
  • Publisher: InTech, Rijeka(in press)(査読有)
• [Book] RNA pathologies in neurological disorders. Neurochemical Mechanisms in Disease, Advances in Neurobiology(Ed by Abel Lajtha) (2011)
  • Author(s): Ohno K, Masuda A
  • Total Pages: 399-415
  • Publisher: Springer, New York
• [Book] Chapter 8 : Molecular defects of acetylcholine receptor subunits in congenital myasthenic syndromes. Pharmacology of Nicotinic Acetylcholine Receptors from the Basic and Therapeutic Perspectives(Ed.By Hugo R.Arias) (2011)
  • Author(s): Ohno K, Engel AG
  • Total Pages: 175-186
  • Publisher: Research Signpost, Kerala