Establishment of treatment of Duchenne muscular dystrophy

• Project/Area Number: 10557076
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 展開研究
• Research Field: Pediatrics
• Research Institution: Kobe University
• Principal Investigator: MATSUO Masafumi Kobe University, School of Medicine, Professor, 医学部, 教授 (10157266)
• Co-Investigator(Kenkyū-buntansha): TAKESHIMA Yasuhiro Kobe University, Hospital, Assistant Professor, 医学部・附属病院, 助手 (40281141)
• Project Period (FY): 1998 – 1999
• Project Status: Completed (Fiscal Year 1999)
• Budget Amount: ¥13,600,000 (Direct Cost: ¥13,600,000); Fiscal Year 1999: ¥3,600,000 (Direct Cost: ¥3,600,000); Fiscal Year 1998: ¥10,000,000 (Direct Cost: ¥10,000,000)
• Keywords: dystrophin / splicing / splicing enhancer sequence / exon skipping / treatment / ジストロフィン遺伝子 / エクソン / スキッピング / アンチセンスオリゴオヌクレオチド

Research Abstract

Duchenne muscular dystrophy (DMD) is a severe muscle wasting disease caused by mutation of the dystrophin gene. DMD patients usually die among the age of 20, but no treatment has been established. We have proposed that out-frame mutation identified in DMD can be corrected to in-frame by inducing exon skipping at the time of splicing. To confirm our proposal, we transfected oligonucleotide which is complementary to splicing enhancer sequence of exon 19 into cultured muscle cells, which was established from DMD case having exon 20 deletion. By this treatment, exon 19 skipping was induced and resulting dystrophin transcript re-gained translational reading frame. Remarkably dystrophin was stained in those transfected cells. This confirms that dystrophin negative cells are able to be converted to dystrophin positive and our proposal is right way to treat DMD.
To expand our strategy, we are currently studying splicing enhancer sequence in other exons, dystrophin, splicing, splicing enhancer sequence, exon skipping, treatment

Research Products

• [Publications] Shiga, N., Matsuo, M., Yokoyama, M and Yokota, Y.: "Study on mutations affecting the muscle promoter/first exon of the dystrophin gene in 92 Japanese dilated cardiomyopathy patients"Am. J. Med. Genet.. 79. 226-227 (1998)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Chen, D.H., Takeshima, Y., Y. Ishikawa, Y. Minami, R and Matsuo, M.: "A novel deletion of the dystrophin S-promoter region consegregating with mental retardation"Neurology. 52. 638-640 (1999)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Surono, A., Takeshima, Y., Wibawa, T., Ikezawa, M., Nonaka, I. and Matsuo, M.: "Circular dystrophin RNAs consisting of exons that were skipped by alternative splicing"Hum. Mol. Genet.. 8. 493-500 (1999)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Patria, S.Y., Takeshima, Y., Suminaga, R., Nakamura, H., Iwasaki, R., Minagawa, T., and Matsuo, M.: "A simple explanation for a case of incompatibility with the reading frame theory in Duchenne muscular dystrophy : failure to detect an aberrant restriction fragment in Southern blot analysis"Brain Dev.. 21. 386-389 (1999)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Dwi Pramono, Z.A., Takeshima, Y., Surono, A., Ishida, T., and Matsuo, M.: "A novel cryptic exon in intron 2 of the human dystrophin gene evolved from an intron by acquiring consensus sequences for splicing at different stages of anthropoid evolution"Biochem. Biophys. Res. Commun.. 267. 321-328 (2000)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Surono, A., Takeshima, Y., Wibawa, T., Pramono, ZA and Matsuo, M.: "Complete skipping of exon 66 due to novel mutations of the dystrophin gene was identified in two Japanese families of DMD with severe mental retardation"Brain Dev.. (in press). (2000)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Shiga,N, Matsuo,M, Yokoyama,M and Yokota,Y.: "Study on mutations affecting the muscle promoter/first exon of the dystrophin gene in 92 Japanese dilated cardiomyopathy patients"Am J Med Genet. 79. 226-227 (1998)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Chen,DH, Takeshima,Y, Ishikawa,Y, Minami,R and Matsuo,M.: "A novel deletion of the dystrophin S-promoter region cosegregating with mental retardation"Neurology. 52. 638-640 (1999)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Surono,A, Takeshima,Y, Wibawa,T, Ikezawa,M, Nonaka,I and Matsuo,M.: "Circular dystrophin RNAs consisting of exons that were skipped by alternative splicing"Hum Mol Genet. 8. 493-500 (1999)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Patria,SY, Takeshima,Y, Suminaga,R, Nakamura,H, Iwasaki,R, Minagawa,T and Matsuo, M.: "A simple explanation for a case of incompatibility with the reading frame theory in Duchenne muscular dystrophy : failure to detect an aberrant restriction fragment in Southern blot analysis"Brain Dev. 21. 386-389 (1999)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Dwi Pramono,ZA, Takeshima,Y, Surono,A, Ishida,T and Matsuo,M.: "A novel cryptic exon in intron 2 of the human dystrophin gene evolved from an intron by acquiring consensus sequence for splicing at different stages of anthropoid evolution"Biochem Biophys Res Commun. 267. 321-328 (2000)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Wibawa,T, Takeshima,Y, Mitsuyoshi,H., Surono,A, Nakamura,H and Matsuo,M.: "Complete skipping of exon 66 due to novel mutation of the dystrophin gene was identified in two Japanese families of DMD with severe mental retardation"Brain Dev. (in press). (2000)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Suminaga,R, Takeshima,Y, Wada,H, Nakamura,H and Matsuo,M.: "Analysis of a nonsense mutation of the alpha-actinin-3 gene in Japanese patients with dystrophinopathy"Am J Med Genet. (in press). (2000)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Surono,A,Takeshima,Y,Wibawa,T,Ikezawa,M,Nonaka,I and Matsuo,M.: "Circular dystrophin RNAs consisting of exons that were skipped by alternative splicing"Hum Mol Genet. 8. 493-500 (1999)
• [Publications] Patria,SY,Takeshima,Y,Suminaga,R,Nakamura,H,Iwasaki,R,Minagawa,T,and Matsuo,M.: "A simple explanation for a case of incompatibility with the reading frame theory in Duchenne muscular dystrophy: failure to detect an aberrant restriction fragment in Southern blot anlysis"Brain Dev. 21. 386-389 (1999)
• [Publications] Dwi Pramono,ZA,Takeshima,Y,Surono,A,Ishida,T and Matsuo,M.: "A novel cryptic exon in intron 2 of the human dystrophin gene evolved from an intron by acquiring consensus sequences for splicing at different stages of anthropoid evolution"Biochem Biophys Res Commun. 267. 321-328 (2000)
• [Publications] Wibata,T,Takeshima,Y,Mitsuyoshi,H.,Surono,A,Nakamura,H and Matsuo,M.: "Complete skipping of exon 66 due to novel mutations of the dystrophin gene was identified in two Japanese families of DMD with severe mental retardation"Brain Dev. (in press). (2000)
• [Publications] Suminaga,R,Takeshima,Y,Wada,H,Nanamura,H and Matsuo,M.: "Analysis of a nonsense mutation of the alpha-actinin-3 gene in Japanese patients with dystrophinopathy"Am J Med Genet. (in press). (2000)
• [Publications] Shiga,N,Matsuo,M,Yokoyama,M,Yokota,Y.: "Study on mutations affecting the muscle promoter/first exon of the dystrophin gene in 92 Japanese dilated cardiomyopathy patients." Am J Med Genet.79. 226-227 (1998)
• [Publications] Surono,A,Takeshima,Y,Wibawa,T,Ikezawa,M,Nonaka,I,Matsuo,M.: "Circular dystrophin RNAs consisting of exons that were skipped by alternative splicing." Hum Mol Genet.8. 493-500 (1999)
• [Publications] Chen,D,Takeshima,Y,Ishikawa,Y,Ishikawa,Y,Minami,R,Matsuo,M.: "A novel deletion of the dystrophin S-promoter region co-segregating with mental retardation." Neurology. (in press). (1999)