Development of therapy for myotonic dystrophy by analyzing splicing mechanism using new splicing quantitative system

• Project/Area Number: 15H06161
• Research Category: Grant-in-Aid for Research Activity Start-up
• Allocation Type: Single-year Grants
• Research Field: Neurology
• Research Institution: The University of Tokyo
• Principal Investigator: YOSHIDA NATSUMI 東京大学, 生命科学ネットワーク, 特任助教 (10760069)
• Project Period (FY): 2015-08-28 – 2017-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000); Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: myotonic dystrophy (DM) / chloride channel (CLCN1) / MBNL / CELF / antisenseoligonucleotide / Tat peptide / CLCN1 / 筋強直性ジストロフィー / AON / TAT / 遺伝子 / 核酸 / 発現制御 / 病理 / 薬学 / 選択的スプライシング / クロライドチャネル

Outline of Final Research Achievements

Muscle hyper-excitability (myotonia) in patients with myotonic dystrophy is caused by the abnormality in alternative splicing of chloride channel (CLCN1). It is important for the treatment to normalize CLCN1 splicing. However, it was found that the genomic structures of mouse and human CLCN1 are considerably different, and we newly detected the abnormal splicing isoform exists in human CLCN1. So, it was necessary to quantify CLCN1 splicing by a new method using human CLCN1 genome. Therefore, we constructed a system to quantify only normal splicing, identify the sequence of antisense oligonucleotide that efficiently normalizes splicing of human CLCN1, and identify the splicing factor for the normalization. We expect that these results will be contributed to development of the treatment.

Research Products

• [Journal Article] Expansions of intronic TTTCA and TTTTA repeats in benign adult familial myoclonic epilepsy (2018)
  • Author(s): Ishiura Hiroyuki、Doi Koichiro、Mitsui Jun、Yoshimura Jun、Matsukawa Miho Kawabe、Takahashi Yuji、Date Hidetoshi、Matsukawa Takashi、Shimizu Jun、Koh Kishin、Takiyama Yoshihisa、Goto Jun、Morishita Shinichi、Tsuji Shoji
  • Journal Title: Nature Genetics Volume: 50 Issue: 4 Pages: 581-590
  • DOI: 10.1038/s41588-018-0067-2
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Splicing of human chloride channel 1 (2015)
  • Author(s): Nakamura, T., Ohsawa-Yoshida, N., Zhao, Y., Koebis, M., Oana, K., Mitsuhashi, H., Ishiura, S.
  • Journal Title: Biochemistry and Biophysics Reports Volume: 5 Pages: 63-69
  • DOI: 10.1016/j.bbrep.2015.11.006
  • Peer Reviewed / Open Access
• [Presentation] マウス膵ベータ細胞株MIN6の飢餓ストレスに対するlncRNAを介した遺伝子発現制御機構の探索 (2017)
  • Author(s): 土屋一郎、荒木海人、石井智子、吉田(大澤)奈摘、太田邦史
  • Organizer: ConBio2017
• [Presentation] 土屋一郎、石井智子、吉田（大澤）奈摘、太田那史 (2016)
  • Author(s): マウス膵β細胞株MIN6における飢餓ストレスに対すlncRNAを介した遺伝子発現制御の探索
  • Organizer: The 39th Annual Meeting of the Molecular Biology Society of Japan
  • Place of Presentation: パシフィコ横浜（神奈川県・横浜市）
  • Year and Date: 2016-11-30
• [Presentation] Identification of interactions between repeat RNAs and RNA binding proteins by the Yeast Three Hybrid system. (2016)
  • Author(s): Natsumi Ohsawa-Yoshida, Josephine GALIPON, Shoichi Ishiura, Kunihiro Ohta.
  • Organizer: The 39th Annual Meeting of the Molecular Biology Society of Japan
• [Presentation] The optimization of antisense oligonucleotide therapy for the abnormality of CLCN1 alternative splicing in DM. (2015)
  • Author(s): Natsumi Ohsawa-Yoshida, Takumi Nakamura, Michinori Koebis, Shoichi Ishiura.
  • Organizer: BMB2015
  • Place of Presentation: 神戸国際会議場（兵庫県・神戸市）
  • Year and Date: 2015-12-01