Molecular bases and their regulations of mRNA aberrations in neuromuscular transmission defects and other muscular diseases

• Project/Area Number: 24390221
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Neurology
• Research Institution: Nagoya University
• Principal Investigator: OHNO Kinji 名古屋大学, 医学(系)研究科(研究院), 教授 (80397455)
• Co-Investigator(Kenkyū-buntansha): MASUDA Akio 名古屋大学, 大学院医学系研究科, 准教授 (10343203) ; ITO Mikako 名古屋大学, 大学院医学系研究科, 助教 (60444402) ; OHKAWARA Bisei 名古屋大学, 高等研究院, 特任講師 (80589606)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥17,940,000 (Direct Cost: ¥13,800,000、Indirect Cost: ¥4,140,000); Fiscal Year 2014: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2013: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2012: ¥7,670,000 (Direct Cost: ¥5,900,000、Indirect Cost: ¥1,770,000)
• Keywords: 先天性筋無力症候群 / 神経筋接合部 / RSPO2 / LGR5 / LRP4 / Wnt / ColQ

Outline of Final Research Achievements

Laser capture microdissection of mouse spinal motor neurons revealed that Rspo2 is highly expressed in spinal motor neurons. Rspo2 induces acetylcholine receptor (AChR) clustering, which is ~80% as potent as agrin. We propose that Rspo2 is an essential AChR clustering-inducing molecule after agrin.
We identified that mutations in LRP4 cause congenital myasthenic syndrome. Mutations in the 3rd beta-propeller domain of LRP4 cause either a defect in neuromuscular signal transmission or a defect in osteogenesis in a position-specific manner.

Research Products

• [Journal Article] Congenital myasthenic syndrome in Japan: ethnically unique mutations in muscle nicotinic acetylcholine receptor subunits (2015)
  • Author(s): Azuma Y, Nakata T, Tanaka M, Shen XM, Ito M, Iwata S, Okuno T, Nomura Y, Ando N, Ishigaki K, Ohkawara B, Masuda A, Natsume J, Kojima S, Sokabe M, Ohno K.
  • Journal Title: Neuromuscul Disord Volume: 25(1) Issue: 1 Pages: 60-69
  • DOI: 10.1016/j.nmd.2014.09.002
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] LRP4 third β-propeller domain mutations cause novel congenital myasthenia by compromising agrin-mediated MuSK signaling in a position-specific manner (2014)
  • Author(s): Ohkawara B, Cabrera-Serrano M, Nakata T, Milone M, Asai N, Ito K, Ito M, Masuda A, Ito Y, Engel AG, Ohno K
  • Journal Title: Hum Mol Genet. Volume: Apr 1;23(7) Issue: 7 Pages: 1856-68
  • DOI: 10.1093/hmg/ddt578
  • Peer Reviewed / Open Access
• [Journal Article] LDB3 splicing abnormalities are specific to skeletal muscles of patients with myotonic dystrophy type 1 and alter its PKC binding affinity (2014)
  • Author(s): Yamashita Y, Matsuura T, Kurosaki T, Amakusa Y, Kinoshita M, Ibi T, Sahashi K, Ohno K.
  • Journal Title: Neurobiol Disord Volume: 69 Pages: 200-205
  • DOI: 10.1016/j.nbd.2014.05.026
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Collagen Q is a key player for developing rational therapy for congenital myasthenia and for dissecting the mechanisms of anti-MuSK myasthenia gravis. (2014)
  • Author(s): Ohno K, Ito M, Kawakami Y, Ohtsuka K.
  • Journal Title: J Mol Neurosci Volume: 53(3) Issue: 3 Pages: 359-361
  • DOI: 10.1007/s12031-013-0170-x
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] HnRNP C, YB-1 and hnRNP L coordinately enhance skipping of human MUSK exon 10 to generate a Wnt-insensitive MuSK isoform (2014)
  • Author(s): Nasrin F, Rahman MA, Masuda A, Ohe K, Takeda J, Ohno K.
  • Journal Title: Sci Rep Volume: 4 Issue: 1 Pages: 6841-6841
  • DOI: 10.1038/srep06841
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Clinical and genetic analysis of the first known Asian family with myotonic dystrophy type 2 (2014)
  • Author(s): Nakayama T, Nakamura H, Oya Y, Kimura T, Imahuku I, Ohno K, Nishino I, Abe K, Matsuura T.
  • Journal Title: J Hum Genet Volume: 59 Issue: 3 Pages: 129-133
  • DOI: 10.1038/jhg.2013.133
  • Peer Reviewed / Open Access
• [Journal Article] A Kir3.4 mutation causes Andersen–Tawil syndrome by an inhibitory effect on Kir2.1. (2014)
  • Author(s): Kokunai Y, Nakata T, Furuta M, Sakata S, Kimura H, Aiba T, Yoshinaga M, Osaki Y, Nakamori M, Itoh H, Sato T, Kubota T, Kadota K, Shindo K, Mochizuki H, Shimizu W, Horie M, Okamura Y, Ohno K, Takahashi MP
  • Journal Title: Neurology Volume: 82 Issue: 12 Pages: 1058-64
  • DOI: 10.1212/wnl.0000000000000239
  • Peer Reviewed / Open Access
• [Journal Article] Exome sequencing of senescence-accelerated mice (SAM) reveals deleterious mutations in degenerative disease-causing genes. (2013)
  • Author(s): Tanisawa K., Mikami E., Fuku N., Honda Y., Honda S., Ohsawa I., Ito M., Endo S., Ihara K., Ohno K., Kishimoto Y., Ishigami A., Maruyama N., Sawabe M., Iseki H., Okazaki Y., Hasegawa-Ishii S., Takei S., Shimada A., Hosokawa M., Mori M., Higuchi K., Takeda T., Higuchi M., Tanaka M.
  • Journal Title: BMC Genomics. Volume: 14(1): Issue: 1 Pages: 248-248
  • DOI: 10.1186/1471-2164-14-248
  • Peer Reviewed / Open Access
• [Journal Article] Mutations in the C-Terminal Domain of ColQ in Endplate Acetylcholinesterase Deficiency Compromise ColQ-MuSK Interaction. (2013)
  • Author(s): Nakata T, Ito M, Azuma Y, Otsuka K, Noguchi Y, Komaki H, Okumura A, Shiraishi K, Masuda A, Natsume J, Kojima S, Ohno K.
  • Journal Title: Human Mutation Volume: in press Issue: 7 Pages: 997-1004
  • DOI: 10.1002/humu.22325
  • Peer Reviewed
• [Journal Article] Gfpt1-myasthenia: Clinical, structural, and electrophysiologic heterogeneity (2013)
  • Author(s): Selcen D, Shen XM, Milone M, Brengman J, Ohno K, Deymeer F, Finkel R, Rowin J, Engel AG.
  • Journal Title: Neurology Volume: 81 Issue: 4 Pages: 378-378
  • DOI: 10.1212/wnl.0b013e31829c5e9c
  • Peer Reviewed
• [Journal Article] HnRNP L and hnRNP LL antagonistically modulate PTB-mediated splicing suppression of CHRNA1 pre-mRNA. (2013)
  • Author(s): Rahman, M. A., Masuda, A., Ohe, K., Ito, M. , Hutchinson, D. O., Mayeda, A., Engel, A. G. and Ohno, K.
  • Journal Title: Sci. Rep. Volume: 3 Issue: 1 Pages: 2931-2931
  • DOI: 10.1038/srep02931
  • Peer Reviewed
• [Journal Article] Clinically applicable antianginal agents suppress osteoblastic transformation of myogenic cells and heterotopic ossifications in mice (2013)
  • Author(s): Yamamoto R, Matsushita M, Kitoh H, Masuda A, Ito M, Katagiri T, Kawai T, Ishiguro N, Ohno K
  • Journal Title: J Bone Miner Metab Volume: 31 Pages: 26-33
  • DOI: 10.1007/s00774-012-0380-2
  • NAID: 10031168834
  • Peer Reviewed
• [Journal Article] S100A10 is required for the organization of actin stress fibers and promotion of cell spreading. (2013)
  • Author(s): Shurovi Sayeed
  • Journal Title: Molecular and cellular biochemistry Volume: 374 Issue: 1-2 Pages: 105-11
  • DOI: 10.1007/s11010-012-1509-2
  • Peer Reviewed
• [Journal Article] Molecular hydrogen attenuates fatty acid uptake and lipid accumulation through downregulating CD36 expression in HepG2 cells (2013)
  • Author(s): Iio A, Ito M, Itoh T, Terazawa R, Fujita Y, Nozawa Y, Ohsawa I, Ohno K, Ito M
  • Journal Title: Med Gas Res Volume: 3 Issue: 1 Pages: 6-6
  • DOI: 10.1186/2045-9912-3-6
  • 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: Sci Rep Volume: 2 Issue: 1 Pages: 209-209
  • DOI: 10.1038/srep00209
  • Peer Reviewed
• [Journal Article] A novel mutation in SCN4A causes severe myotonia and school-age-onset paralytic episodes (2012)
  • Author(s): Yoshinaga Y. Sakoda S-I, Good JM, Takahashi MP, Kubota T, Arikawa-Hirasawa E, Nakata T, Ohno K, Kitamura T, Kobayashi K, Ohtsuka Y
  • Journal Title: J Neurol Sci Volume: 315(1-2) Issue: 1-2 Pages: 15-9
  • DOI: 10.1016/j.jns.2011.12.015
  • 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) Issue: 3 Pages: 219-220
  • DOI: 10.1038/jhg.2011.152
  • 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) Issue: 7 Pages: 1384-1392
  • DOI: 10.1038/mt.2012.34
  • Peer Reviewed
• [Journal Article] Drinking hydrogen water and intermittent hydrogen gas exposure, but not lactulose or continuous hydrogen gas exposure, prevent 6-hydorxydopamine-induced Parkinson's disease in rats (2012)
  • Author(s): Ito M, Hirayama M, Yamai K, Goto S, Ito M, Ichihara M, Ohno K
  • Journal Title: Med Gas Res Volume: 2 Issue: 1 Pages: 15-15
  • DOI: 10.1186/2045-9912-2-15
  • Peer Reviewed
• [Journal Article] The unstable CCTG repeat responsible for myotonic dystrophy type 2 originates from an AluSx element insertion into an early primate genome (2012)
  • Author(s): Kurosaki, T., Ueda, S., Ishida, T., Abe, K., Ohno, K. and Matsuura, T
  • Journal Title: PLos One Volume: 7 Issue: 6 Pages: e38379-e38379
  • DOI: 10.1371/journal.pone.0038379
  • Peer Reviewed
• [Journal Article] Four parameters increase the sensitivity and specificity of the exon array analysis and disclose twenty-five novel aberrantly spliced exons in myotonic dystrophy. (2012)
  • Author(s): Yamashita Y, Matsuura T, Shinmi J, Amakusa Y, Masuda A, Ito M, Kinoshita M, Furuya H, Abe K, Ibi T, Sahashi K, Ohno K.
  • Journal Title: Journal of Human Genetics Volume: 57 Issue: 6 Pages: 368-374
  • DOI: 10.1038/jhg.2012.37
  • Peer Reviewed
• [Journal Article] Position-dependent FUS-RNA interactions regulate alternative splicing events and transcriptions. (2012)
  • Author(s): Ishigaki S, Masuda A, Fujioka Y, Iguchi Y, Katsuno M, Shibata A, Urano F, Sobue G, Ohno K
  • Journal Title: Sci Rep Volume: 2 Issue: 1 Pages: 529-529
  • DOI: 10.1038/srep00529
  • Peer Reviewed
• [Presentation] Alternative splicing of human MUSK exon 10 is physiologically regulated by multiple splicing regulatory cis-elements and cognate trans-factors (2014)
  • Author(s): Rahman MA, Masuda A, Takeda J, Ohno K
  • Organizer: RNA Biology meeting, Cold Spring Harbor Asia Conference
  • Place of Presentation: Suzhou, China
  • Year and Date: 2014-11-10 – 2014-11-14
• [Presentation] IntSplice: A tool to predict aberrant splicing of an SNV at intronic positions -50 to -3 (2014)
  • Author(s): Ohno K, Shibata A, Okuno T, Rahman MA, Azuma Y, Masuda A
  • Organizer: 64th Annual Meeting of the American Society of Human Genetics
  • Place of Presentation: San Diego, California, USA
  • Year and Date: 2014-10-18 – 2014-10-22
• [Presentation] Maintenance of the neuromuscular junction and its aberrations in hereditary and autoimmune disorders (2014)
  • Author(s): Ohno K
  • Organizer: Guarda-Symposium 2014 on the Molecular and Cell Biology of the Neuromuscular System
  • Place of Presentation: Guarda, Switzerland
  • Year and Date: 2014-09-01
  • Invited
• [Presentation] Collagen Q is a key player for developing rational therapy for congenital myasthenia and for causing anti-MuSK myasthenia gravis (2013)
  • Author(s): Ohno K, Ito M, Kawakami Y, Ohtsuka K, Krejci E.
  • Organizer: XIV International Symposium on Cholinergic Mechanisms (Platform)
  • Place of Presentation: Hangzhou, China
  • Invited
• [Presentation] Mutations in the third β-propeller domain of LRP4 in congenital myasthenia compromise agrin-mediated MuSK signaling in a position-specific manner (2013)
  • Author(s): Ohkawara B, Cabrera M, Nakata T, Milone M, Ito Y, Engel AG, Ohno K
  • Organizer: 36th Annual Meeting of the Japan Neuroscience Society (Poster)
  • Place of Presentation: 国立京都国際会館(京都市)
• [Presentation] R-spondin2 is crucial for neuromuscular junction formation (2013)
  • Author(s): Nakashima H, Ohkawara B, Imagama S, Ito Z, Ishiguro N, Ohno K
  • Organizer: 43rd Annual Meeting, Society for Neuroscience (Poster)
  • Place of Presentation: San Diego, USA
• [Presentation] Mutations in LRP4 in congenital myasthenia reveal position-specific regulations of agrin and Wnt signaling of LPR4 (2013)
  • Author(s): Ohkawara B, Cabrera M, Nakata T, Shen X, Ito Y, Engel AG, Ohno K
  • Organizer: 43rd Annual Meeting, Society for Neuroscience (Poster)
  • Place of Presentation: San Diego, USA
• [Presentation] Alternative RNA splicing regulating the molecular architecture of NMJ (2013)
  • Author(s): Nasrin F, Rahman MA, Ohe K, Masuda A, Ohno K
  • Organizer: 5th NAGOYA Global Retreat (Poster)
  • Place of Presentation: あいち健康プラザ（名古屋市）
• [Presentation] RNA diseases caused by defects in cis-acting elements and trans-acting factors (2013)
  • Author(s): Ohno K
  • Organizer: "Danish-Japanese joint seminar on aberrant RNA splicing in neuromuscular disease", Department of Biochemistry and Molecular Biology, University of Southern Denmark (SDU)
  • Place of Presentation: Odense, Denmark
  • Invited
• [Presentation] Investigation of 11 Patients with GFPT1-Myasthenia Reveals Clinical, Structural, and Electrophysiologic Heterogeneity (2013)
  • Author(s): Selcen D, Shen X-M, Milone M, Brengman J, Ohno K, McQuillen M, Deymeer F, Finkel R, Rowin J, Engel AG
  • Organizer: 65th American Academy of Neurology (Platform)
  • Place of Presentation: San Diego, USA
• [Presentation] Molecular mechanism of the inhibitory effect of hydrogen on inflammation (2013)
  • Author(s): Ito M, Itoh T, Ohno K, Nozawa Y
  • Organizer: 90th Annual Meeting of the Physiological Society of Japan (Platform)
  • Place of Presentation: タワーホール船堀（東京都江戸川区）
• [Presentation] Specific binding of collagen Q to the neuromuscular junction is exploited to cure congenital myasthenia and to explore bases of myasthenia gravis (2012)
  • Author(s): Ohno K
  • Organizer: 11th International Meeting on Cholinesterases
  • Place of Presentation: Kazan, Russia
  • Invited
• [Presentation] EMG findings at non-myotonia and myotonia states in patients with Na channelopathies (2012)
  • Author(s): Kinoshita M, Kokunai Y, Kubota T, Takahashi M, Sasaki R, Ohno K, Hirose K
  • Organizer: The 11th Annual Scientific Meeting of the Asian and Oceanian Myology Center (Poster)
  • Place of Presentation: 京都市芝蘭会館（京都市）
• [Presentation] In myotonic dystrophy type 1 Peroxiredoxin-2 of RBC membrane protein was reduced, compared with a normal subject (2012)
  • Author(s): Kinoshita M, Morita S, Ohno K, Hirose K
  • Organizer: The 11th Annual Scientific Meeting of the Asian and Oceanian Myology Center (Poster)
  • Place of Presentation: 京都市芝蘭会館（京都市）
• [Presentation] Global analysis of RNA-binding sites of CUGBP1 and MBNL1 reveal their preferential binding to 3’ UTRs to regulate mRNA stability (2012)
  • Author(s): Masuda A, Andersen HS, Doktor TK, Okamoto T, Ito I, Andresen BS, Ohno K
  • Organizer: The 11th Annual Scientific Meeting of the Asian and Oceanian Myology Center (Poster)
  • Place of Presentation: 京都市芝蘭会館（京都市）
• [Presentation] Functional characterization of GFPT1 during myogenesis and NMJ formation (2012)
  • Author(s): Ohkawara B, Fuse Y, Nakashima H, Ohno K
  • Organizer: The 11th Annual Scientific Meeting of the Asian and Oceanian Myology Center (Poster)
  • Place of Presentation: 京都市芝蘭会館（京都市）
• [Presentation] Congenital defects of neuromuscular signal transduction (2012)
  • Author(s): Ohno K
  • Organizer: 3rd Berlin Summer School for Myology
  • Place of Presentation: Berlin, Germany
  • Invited
• [Presentation] Molecular bases and therapeutic intervention of neuromuscular transmission defects (2012)
  • Author(s): Ohno K
  • Organizer: Ninth French-Japanese Workshop on Muscular Dystrophies
  • Place of Presentation: ＪＡ共済ビル カンファレンス・ホール（東京都千代田区）
  • Invited
• [Presentation] Global mapping and global expression profiling of RNA-binding proteins that are associated with neurological and neuromuscular diseases (2012)
  • Author(s): Ohno K
  • Organizer: 35th Annual Meeting of the Japan Neuroscience Society
  • Place of Presentation: 名古屋国際会議場（名古屋市）
  • Invited
• [Presentation] Mutation in congenital myasthenic syndrome reveals opposite splicing regulatory effects of hnRNPs L and LL (2012)
  • Author(s): Rahman MA, Ohe K, Masuda A, Ito M, Hutchinson DO, Engel AG, Ohno K
  • Organizer: 35th Annual Meeting of the Japan Neuroscience Society (Poster)
  • Place of Presentation: 名古屋国際会議場（名古屋市）
• [Presentation] Global mapping of FUS-binding sites and global profiling of FUS-mediated RNA metabolisms in the mouse brain (2012)
  • Author(s): Masuda A, Ishigaki S, Fujioka Y, Iguchi Y, Katsuno M, Shibata A, Urano F, Sobue G, Ohno K
  • Organizer: 62nd Annual Meeting of the American Society of Human Genetics (Poster)
  • Place of Presentation: San Francisco, USA
• [Presentation] A comprehensive and efficient algorithm to predict splicing consequences of intronic nucleotide substitutions in the human genome (2012)
  • Author(s): Shibata A, Masuda A, Ohno K
  • Organizer: 62nd Annual Meeting of the American Society of Human Genetics (Poster)
  • Place of Presentation: San Francisco, USA
• [Presentation] Molecular hydrogen alters signaling pathways and gene expression profiles in multiple mouse organs (2012)
  • Author(s): Sobue S, Yamai K, Ito M, Ohno K, Ito M, Ohkuwa T, Ichihara M
  • Organizer: 52nd Annual Meeting of the American Society for Cell Biology (Poster)
  • Place of Presentation: San Francisco, USA
• [Book] Molecular Genetics of Congenital Myasthenic Syndromes. in eLS. DOI: 10.1002/9780470015902.a0024314) (2014)
  • Author(s): Ohno K, Ohkawara B, Ito M, Engel AG.
  • Publisher: John Wiley & Sons, Inc., Hoboken, Chichester
  • DOI: 10.1002/9780470015902.a0024314
• [Book] "Collagen Q is a key player for developing rational therapy for congenital myasthenia and for dissecting the mechanisms of anti-MuSK myasthenia gravis" in J Mol Neurosci (2014)
  • Author(s): Ohno K, Ito M, Kawakami Y.
  • Publisher: Springer, New York
• [Book] "Molecular Genetics of Congenital Myasthenic Syndromes" in eLS (2014)
  • Author(s): Ohno K, Ohkawara B, Ito M, Engel AG.
  • Publisher: John Wiley & Sons, Inc., Hoboken
• [Book] "Specific binding of collagen Q to the neuromuscular junction is exploited to cure congenital myasthenia and to explore bases of myasthenia gravis" in Chem Biol Interact 203(1) (2013)
  • Author(s): Ohno K, Ito M, Kawakami Y, Krejci E, Engel AG.
  • Total Pages: 376
  • Publisher: Elsevier, Amsterdam
• [Book] Specific binding of collagen Q to the neuromuscular junction is exploited to cure congenital myasthenia and to explore bases of myasthenia gravis. Chem Biol Interact 203(1) (2013)
  • Author(s): Ohno K, Ito M, Kawakami Y, Krejci E, Engel AG.
  • Total Pages: 376
  • Publisher: Elsevier, Amsterdam
• [Book] Chapter 2: Intronic and exonic nucleotide variations that affect RNA splicing in humans. Genomics I - Humans, Animals and Plants. (2013)
  • Author(s): Ohe K, Masuda A, Ohno K.
  • Total Pages: 344
  • Publisher: iConcept Press, Hong Kong
• [Book] Molecular Hydrogen as an Emerging Therapeutic Medical Gas for Neurodegenerative and Other Diseases. Oxid Med Cell Longev vol.2012. Article ID:353152 (2012)
  • Author(s): Ohno K, Ito M, Ichihara M, and Ito M.
  • Publisher: Hindawi Publishing Corporation, Cairo
• [Patent(Industrial Property Rights)] 骨格筋増量剤及びその用途 (2014)
  • Inventor(s): 大野欽司、石黒直樹、飛田哲朗、松下雅樹
  • Industrial Property Rights Holder: 国立大学法人名古屋大学
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2014-176248
  • Filing Date: 2014-08-29
• [Patent(Industrial Property Rights)] 「骨系統疾患治療薬及びその用途」 (2013)
  • Inventor(s): 大野欽司、石黒直樹、鬼頭浩史、松下雅樹
  • Industrial Property Rights Holder: 国立大学法人名古屋大学
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2013-047426
  • Filing Date: 2013-03-10