Study on the treatment of Duchenne musclar dystrophy with chimera RNA/DNA

• Project/Area Number: 13307026
• Research Category: Grant-in-Aid for Scientific Research (A)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Pediatrics
• Research Institution: Kobe University
• Principal Investigator: MATSUO Masafumi Kobe University, Graduate School of Medicine, Professor, 大学院・医学系研究科, 教授 (10157266)
• Co-Investigator(Kenkyū-buntansha): YAGI Mariko Kobe University, Hospital, Research Associate, 医学部付属病院, 助手 (60362787) ; TAKESHIMA Tasuhiro Kobe University, Graduate School of Medicine, Associate Professor, 大学院・医学系研究科, 助教授 (40281141)
• Project Period (FY): 2001 – 2003
• Project Status: Completed (Fiscal Year 2003)
• Budget Amount: ¥55,120,000 (Direct Cost: ¥42,400,000、Indirect Cost: ¥12,720,000); Fiscal Year 2003: ¥16,120,000 (Direct Cost: ¥12,400,000、Indirect Cost: ¥3,720,000); Fiscal Year 2002: ¥16,120,000 (Direct Cost: ¥12,400,000、Indirect Cost: ¥3,720,000); Fiscal Year 2001: ¥22,880,000 (Direct Cost: ¥17,600,000、Indirect Cost: ¥5,280,000)
• Keywords: dystrophin / Duchenne / muscular dystrophy / molecular therapy / 遺伝子異常 / デュシェンヌ型筋ジストロフィー / キメラRNA / DNA / 点変異 / 筋細胞 / 治療 / 点突然変異

Research Abstract

In this study the method to treat Duchenne musclar dystrophy (DMD) was examined by inducing exon 41 skipping. In one DMD a nonsense mutation was identified in exon 41 of the dystrophin gene. It was supposed that he will start dystrophin production when exon 41 skipping was induced resulting in in-frame mRNA. RNA/ENA chimera was employed to induce exon 41 skipping. Several RNA/ENA chimera were designed to cover the full-length of exon 41 and each of RNA/ENA chimera examined for its activity to induce exon 41 skipping. As a result the best RNA/ENA was identified. The selected RNA/ENA chimera was transfected to myocytes of the DMD case. Remarkably, exon 41 skipping was induced in his dystrophin mRNA and dystrophin expression was shown.

Research Products

• [Publications] Yagi M: "Two alternative exons can result from activation of the cryptic splice acceptor site deep within intron 2 of the dystrophin gene in a patient with as yet asymptomatic dystrophinopathy"Hum.Gemet.. 112. 164-170 (2003)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Ito T: "Analysis of dystrophin mRNA from skeletal muscle but not from lymphocytes led to identification of a novel nonsense mutation in a carrier of Duchenne muscular dystrophy"J.Neurol. 250. 581-587 (2003)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Adachi K: "Heterogpus dystrophin mRNAs produced by a novel splice acceptor site mutation in intermediate dystrophinopathy"Ped.Res.. 53. 1-7 (2003)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Masafumi M: "Treatment of Duchenne muscular dystrophy with oligonucleotides against an exonic splicing enhancer sequence"Basic and Applied Myology. (in press). (2003)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Nakayama Y: "Cloning of cDNA encoding a regeneration-associated muscle protease whose expression is attenuated in cell lines derived from Duchenne muscular dystrophy patients"American Journal of Pathology. (in press). (2004)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Yagi M, Takeshima Y, Wada H, Nakamura H, Matsuo M.: "Two alternative exons can result from activation of the cryptic splice acceptor site deep within intron 2 of the dystrophin gene in a patient with as yet asymptomatic dystrophinopathy."Hum.Genet.. 112. 164-170 (2003)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Ito T, Takeshima Y, Yagi M, Kamei S, Wada H, Matsuo M.: "Analysis of dystrophin mRNA from skeletal muscle but not from lymphocytes led to identification of a novel nonsense mutation in a carrier of Duchenne muscular dystrophy."J.Neurol.. 250. 581-587 (2003)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Adachi K, Takeshima Y, Wada H, Yagi Y, Nakamura H, Matsuo M.: "Hetergpus dystrophin mRNAs produced by a novel splice acceptor site mutation in intermediate dystrophinopathy."Ped.Res.. 53. 1-7 (2003)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Matsuo M, Yagi M, Takeshima Y.: "Treatment of Duchenne muscular dystrophy with oligonucleotides against an exonic splicing enhancer sequence."Basic and Applied Myology. (in press). (2003)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Nakayama Y, Nara N, Kawakita Y, Takeshima Y, Arakawa M, Katoh M, Morita S, Iwatsuki K, Tanaka K, Okamoto S, Kitamura T, Seki N, Matsuda R, Matsuo M, Saito K, Hara T.: "Cloning of cDNA Encoding a Regeneration-associated Muscle Protease Whose Expression is Attenuated in Cell Lines Derived from Duchenne Muscular Dystrophy Patients"American Journal of Pathology. (in press). (2004)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Yagi M: "Two alternative exons can result from activation of the cryptic splice acceptor site deep within intron 2 of the dystrophin gene in a patient with as yet asymptomatic dystrophinopathy"Hum.Gemet.. 112. 164-170 (2003)
• [Publications] Ito T: "Analysis of dystrophin mRNA from skeletal muscle but not from lymphocytes led to identification of a novel nonsense mutation in a carrier of Duchenne muscular dystrophy"J.Neurol. 250. 581-587 (2003)
• [Publications] Adachi K: "Hetergpus dystrophin mRNAs produced by a novel splice acceptor site mutation in intermediate dystrophinopathy"Ped.Res.. 53. 1-7 (2003)
• [Publications] Masafumi M: "Treatment of Duchenne muscular dystrophy with oligonucleotides against an exonic splicing enhancer sequence"Basic and Applied Myology. in press. (2003)
• [Publications] Nakayama Y: "Cloning of cDNA Encoding a Regeneration-associated Muscle Protease Whose Expression is Attenuated in Cell Lines Derived from Duchenne Muscular Dystrophy Patients"American Journal of Pathology. in press. (2004)
• [Publications] Saito-Ohara F, Fukuda Y, Ito M, Agarwala K, Hayashi M, Matsuo M, Imoto I, Yamakawa K, Nakamura Y, Inazawa J.: "The Xq22 inversion breakpoint interrupted a novel Ras-like GTPase gene in a patient with Duchenne muscular dystrophy and profound mental retardation"Am J Hum Genet.. 71. 637-645 (2002)
• [Publications] Suminaga R, Takeshima Y, Adachi K, Yagi M, Nakamura H, Matsuo M.: "A novel cryptic exon in intron III of the dystrophin gene was incorporated into dystrophin mRNA with a single nucleotide deletion in exon 5"J Hum Genet.. 47. 196-201 (2002)
• [Publications] Matsuo M.: "Duchenne and Becker muscular dystrophy : from gene diagnosis to molecular therapy"IUBMB Life. 53. 147-152 (2002)
• [Publications] Adachi K, Takeshima Y, Wada H, Yagi M, Nakamura H, Matsuo M.: "Heterogous dystrophin mRNA produced by a novel splice acceptor site mutation in intermediate dystrophinopathy"Pediatr Res.. 53. 125-131 (2003)
• [Publications] Yagi M, Takeshima Y, Nakamura H, Matsuo M.: "Two alternative exons can result from activation of the cryptic splice acceptor site deep within intron 2 of the dystrophin gene in a patient with as yet asymptomatic dystrophinopathy"Hum Genet.. 112. 164-170 (2003)
• [Publications] Ito T.: "One of three examined purine-rich sequences selected from dystrophin exons exhibits splicing enhancer activity"Acta Myologica. 2. 151-153 (2001)
• [Publications] Matsuo M.: "Treatment of Duchenne muscular dystrophy at the mRDA level."Frontiers in human genetics diseases and technologies.. 347-361 (2001)
• [Publications] Tachi N.: "Fukuyama muscular dystrophy associated with lack of C-terminal domain of dystrophin."Pediatr. Neurol. 24. 373-378 (2001)
• [Publications] Takeshima Y.: "Nakamura, H, Matsuo, M. Oligonucleotides against a splicing enhancer sequence led to dystrophin production in muscle cells from a Duchenne muscular dystrophy patient."Brain Dev.. 23. 788-798 (2001)
• [Publications] Ito T.: "Purine-rich exon sequences are not necessarily splicing enhancer sequence in the dystrophin gene."Kobe. J. Med. Sci.. 47. 193-202 (2001)