Gene therapy for progressive muscular dystrophy using a new generation adenovirus vector (gutless adenovirus)

• Project/Area Number: 15590897
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Neurology
• Research Institution: Kumamoto University
• Principal Investigator: UCHIN Makoto Kumamoto University, Department of Neurology, Professor, 大学院・医学薬学研究部, 教授 (20117336)
• Co-Investigator(Kenkyū-buntansha): MAEDA Yasushi Kumamoto University Hospital, Department of Neurology, assistant, 医学部附属病院, 助手 (60346997)
• Project Period (FY): 2003 – 2004
• Project Status: Completed (Fiscal Year 2004)
• Budget Amount: ¥3,500,000 (Direct Cost: ¥3,500,000); Fiscal Year 2004: ¥1,700,000 (Direct Cost: ¥1,700,000); Fiscal Year 2003: ¥1,800,000 (Direct Cost: ¥1,800,000)
• Keywords: gene therapy / adenovirus vector / helper-dependent / muscular dystrophy / attachment receptor / dystrophin / 反復投与 / ヘルパーウイルス依存型アデノウイルスベクター / CAR / mdxマウス / ヌードmdxマウス

Research Abstract

The helper-dependent adenovirus(HDAd) vector is less immunogenic and has a larger cloning capacity of up to 37 kb enough to carry the full-length dystrophin cDNA. However, high and long-term expression of dystrophin transduced to mature muscle still remains difficult. One of the main reasons for this is that the expression of the Coxsakievirus and adenovirus receptor(CAR) is very low in mature muscle. We have constructed two different HDAd vectors. One contains the LacZ and the murine full-length dystrophin expression cassette(HDAdLacZ-dys), and the other is a new, improved vector containing the CAR and the dystrophin expression cassette(HDAdCAR-dys). We initially demonstrated high dystrophin expression and prevention of the dystrophic pathology in mdx muscle injected during the neonatal phase with HDAdLacZ-dys. Furthermore, we demonstrated that repeated injections of HDAdCAR-dys into mature muscle led to approximately nine times greater dystrophin-positive fibers in number than a single injection, thereby recovering the expression of dystrophin-associated proteins. This data has also shown that HDAdCAR-dys enabled administration of adenovirus(Ad) vector to the host with pre-existing immunity to the same serotype of Ad. Repetitive injections of the HDAd vector containing the CAR and the dystrophin expression cassette could improve the efficiency of subsequent dystrophin gene transfer to mature mdx muscle. This result suggests that our new HDAd vector will provide a novel gene therapy strategy for Duchenne muscular dystrophy, raising the prospects for gene therapy of other hereditary myopaties.

Research Products

• [Journal Article] Myopathy phenotype in transgenic mice expressing mutated PABPN1 as a model of oculopharyngeal muscular dystrophy (2004)
  • Author(s): Hino H., Araki K., Uyama E., Takeya M., Araki M., Yoshinobu K., Miike K., Kawazoe Y., Maeda Y., Uchino M., Yamamura K.
  • Journal Title: Hum.Mol.Genet. 13 Pages: 181-190
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Myopathy phenotype in transgenic mice expressing mutated PABPN1 as a model of oculopharyngeal muscular dystrophy. (2004)
  • Author(s): Hino H., Araki K., Uyama E., Takeya M., Araki M., Yoshinobu K., Miike K., Kawazoe Y., Maeda Y., Uchino M., Yamamura K.
  • Journal Title: Hum.Mol.Genet. 13 Pages: 181-190
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Targeted conversion of the transthyretin gene in vitro and in vivo. (2004)
  • Author(s): Nakamura M., Ando Y., Nagahara S., Sano A., Ochiya T., Maeda S., Kawaji T., Ogawa M., Hirata A., Terazaki H., Haraoka K., Tanihara H., Ueda M., Uchino M., Yamamura K., Yamashita T.
  • Journal Title: Gene Therapy 11 Pages: 838-846
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Effective repetitive dystrophin gene transfer into skeletal muscle of adult mdx mice using a helper-dependent adenovirus vector expressing the Coxsackie-virus and adenovirus receptor (CAR) and dystrophin.
  • Author(s): Uchida Y., Maeda Y, Kimura E, Yamashita S, Nishida Y, Arima T, Hirano T, Uyama E, Mita S., Uchino M.
  • Journal Title: J Gene Med (in press)
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Effective repetitive dystrophin gene transfer into skeletal muscle of adult mdx mice using a helper-dependent adenovirus vector expressing the Coxsackievirus and adenovirus receptor(CAR) and dystrophin.
  • Author(s): Uchida Y., Maeda Y, Kimura E, Yamashita S, Nishida Y, Arima T, Hirano T, Uyama E, Mita S., Uchino M.
  • Journal Title: J Gene Med (in press)
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Effective repetitive dystrophin gene transfer into skeletal muscle of adult mdx mice using a helper-dependent adenovirus vector expressing the Coxsackie-virus and adenovirus receptor (CAR) and dystrophin.
  • Author(s): Uchida Y., Maeda Y, Kimura E, Yamashita S, Nishida Y, Arima T, Hirano T, Uyama E, Mita S., Uchino M.
  • Journal Title: J Gene Med (in press)
• [Publications] Maeda Y., Uchino M.et al.: "Cre/lox P-mediated adenovirus type 5 packaging signal excision demonstrates that core element VI is sufficient for virus packaging"Virology. 309. 330-338 (2003)