U12依存性スプライシングとALSサーキットパソロジー

• Project Area: Generation of synapse-neurocircuit pathology
• Project/Area Number: 25110714
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Niigata University
• Principal Investigator: 小野寺 理 新潟大学, 脳研究所, 教授 (20303167)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Project Status: Discontinued (Fiscal Year 2014)
• Budget Amount: ¥15,600,000 (Direct Cost: ¥12,000,000、Indirect Cost: ¥3,600,000); Fiscal Year 2014: ¥7,800,000 (Direct Cost: ¥6,000,000、Indirect Cost: ¥1,800,000); Fiscal Year 2013: ¥7,800,000 (Direct Cost: ¥6,000,000、Indirect Cost: ¥1,800,000)
• Keywords: 神経病態

Outline of Annual Research Achievements

ALSのサーキットパソロジーには次の2点1) TDP-43病理像の錐体路への選択性，2)細胞死の運動神経への選択性の解明が必要であり，各々について研究を進めた．本年度は，引き続きTDP-43病理像の錐体路への選択性についてて検討した．TDP-43遺伝子変異，及び C9ORF72変異によるALS／FTLDの発見は，組織への選択性を規定する因子が，遺伝子ではなく，侵される細胞側の特性にあることを示す．その細胞側の特性として，我々は細胞でのTDP-43 mRNA の制御機構の相違を考え，これを検討した．これに示唆を与える事実として，まずTDP-43は自己量の制御機構をもち，最終エクソンがその制御の主体を担っている．同部は複数のスプライシング部位とpolyA結合部位をもつ．TDP-43 mRNA は核内に多量に存在する．また制御機構の乱れを示唆する所見として，疾患関連変異が最終エクソンに集中する．TDP-43変異を持つ患者由来人工多能性幹細胞由来の運動神経細胞ではTDP-43の増加が示唆されていることがあげられる．そこでTDP-43自身のｍRNAの脊髄運動神経細胞での検討を行った．神経細胞でのTDP-43 mRNAの細胞内での核，細胞質における存在比率と，を検討した．in situ hybridization 法を用い高感度に，かつ定量的にTDP-43 mRNAを測定できる系を開発し，本方法による検討を行った．その結果ALS脊髄運動神経細胞においてTDP-43 mRNAの細胞内局在の変化を同定した．本発見は細胞毎のTDP-43量調節について検討を加える基盤となる．

Research Progress Status

26年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

26年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] A blinded international study on the reliability of genetic testing for GGGGCC-repeat expansions in C9orf72 reveals marked differences in results among 14 laboratories. (2014)
  • Author(s): AkimotoC, Onodera O ら
  • Journal Title: J Med Genet. Volume: 51 Issue: 6 Pages: 419-424
  • DOI: 10.1136/jmedgenet-2014-102360
  • Peer Reviewed
• [Journal Article] Bunina bodies in motor and non-motor neurons revisited: A pathological study of an ALS patient after long-term survival on a respirator. (2014)
  • Author(s): Kimura T, Jiang H, Konno T, Seto M, Iwanaga K, Tsujihata M, Satoh A, Onodera O, Kakita A, Takahashi H.
  • Journal Title: Neuropathology Volume: なし Issue: 4 Pages: 392-397
  • DOI: 10.1111/neup.12105
  • Peer Reviewed / Open Access
• [Journal Article] Minor splicing pathway is not minor any more: implications for the pathogenesis of motor neuron diseases. (2014)
  • Author(s): Onodera O, Ishihara T, Shiga A, Ariizumi Y, Yokoseki A, Nishizawa M.
  • Journal Title: Neuropathology Volume: 34 Issue: 1 Pages: 99-107
  • DOI: 10.1111/neup.12070
  • Peer Reviewed
• [Journal Article] ERBB4 Mutations that Disrupt the Neuregulin-ErbB4 Pathway Cause Amyotrophic Lateral Sclerosis Type 19. (2013)
  • Author(s): Takahashi Y, Fukuda Y, Yoshimura J, Toyoda A, (33名省略）, Morishita S, Goto J and Tsuji S.
  • Journal Title: Am J Hum Genet Volume: 93 Issue: 5 Pages: 900-905
  • DOI: 10.1016/j.ajhg.2013.09.008
  • Peer Reviewed
• [Journal Article] Sporadic ALS with compound heterozygous mutation in the SQSTM1 gene. (2013)
  • Author(s): Shimizu H, Toyoshima Y, Shiga A, Yokoseki A, Arakawa K, Sekine Y, Shimohata T, Ikeuchi T, Nishizawa M, Kakita A, Onodera O, Takahashi H
  • Journal Title: Acta Neuropathol Volume: 126 Issue: 3 Pages: 453-459
  • DOI: 10.1007/s00401-013-1150-5
  • Peer Reviewed
• [Journal Article] Decreased number of Gemini of coiled bodies and U12 snRNA level in amyotrophic lateral sclerosis. (2013)
  • Author(s): Ishihara T, Ariizumi Y, Shiga A, et al ;
  • Journal Title: Human Molecular Genetics Volume: 22 Issue: 20 Pages: 4136-4147
  • DOI: 10.1093/hmg/ddt262
  • Peer Reviewed
• [Presentation] The reduction of minor spliceosome and aberrant splicing in ALS. (2014)
  • Author(s): Tomohiko Ishihara,Akihide Koyama, Masatoyo Nishizawa, Osamu Onodera
  • Organizer: Cell symposia Regulatory RNAs
  • Place of Presentation: Claremont Hotel & Spa, USA
  • Year and Date: 2014-10-20
• [Presentation] Endogenous TDP-43 over-expression model with disrupted auto-regulation (2014)
  • Author(s): Akihiro Sugai, Taisuke Kato, Akihide Koyama, Masatoyo Nishizawa, Osamu Onodera
  • Organizer: 10th Annual Meeting of the Oligonucleotide Therapeutics Society
  • Place of Presentation: Hilton San Diego Resort & Spa, USA
  • Year and Date: 2014-10-12
• [Presentation] Update in amyotrophic lateral sclerosis (2014)
  • Author(s): Osamu Onodera, Masatoyo Nishizawa, Hitoshi Takahashi
  • Organizer: XVIII International congress of neuropathology.
  • Place of Presentation: Sheratom Rio Hotel, Rio de Janeiro,Brazil
  • Year and Date: 2014-09-15
  • Invited
• [Presentation] Alternation of miRNA expression in TDP-43 depleted spinal motor neuron (2014)
  • Author(s): Atsushi Shiga, Chigusa Kondo, Sachiko Hirokawa, Akinori Miyashita, Toshiya Sato, Kenji Sakimura, Masatoyo Nishizawa, Osamu Onodera
  • Organizer: Neuroscience 2014
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2014-09-13
• [Presentation] RNA metabolism and ALS (2014)
  • Author(s): Osamu Onodera
  • Organizer: Neuroscience 2014
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2014-09-12
  • Invited
• [Presentation] Analysis of intracellular distribution of TDP-43 mRNA in affected spinal motor neuron with ALS (2014)
  • Author(s): Taisuke Kato, Akihide Koyama, Akihiro Sugai, Yasuko Toyoshima, Akiyoshi Kakita, Hitoshi Takahashi, Osamu Onodera
  • Organizer: Neuroscience 2014
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2014-09-11
• [Presentation] TDP-43 is autoregulated by multiple excisions of introns in exon6 and reservation of mRNA in nucleus by TDP-43 (2014)
  • Author(s): Akihide Koyama, Akihiro Sugai, Taisuke Kato, Takuya Konno, Tomohiko Ishihara, Masatoyo Nishizawa, Osamu Onodera
  • Organizer: Neuroscience 2014
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2014-09-11