Duchenne muscular dystrophies (DMD) and Limb-girdle muscular dystrophies (LGMD) are common inherited degenerative muscle diseases caused by mutations in genes coding for memberance associated proteins in muscle cells. DMD and LGMD often manifest themselves in young age and lead to severe morbidity and fatality, with no currently available effective treatment. In addition, the diseases are usually genotypically recessive, which makes them suitable for gene replacement therapy with vectors. Recombinant adeno-associated virus (rAAV) is one such vector based on defective human parvoviruses. rAAV system has attracted attention due to its non-pathogenicity, genomic integration, transduction of quiescent cells, and apparent lack of cellular immune reactions. In contrast to other viral vectors, rAAV is capable of efficiently bypassing the myofiber basal lamina and tranducing mature muscle cells. We have demonstrated that rAAV vectors harboring a foreign gene can achieve highly efficient and sustained gene transfer in mature muscle of immunocompetent animals for more than 1.5 years without detectable toxicity. Recently, significant improvement in vector production methodology has made it possible to generate high titer and high quality rAAV vectors completely free of helper adenovirus contamination. However, no experiments using rAAV vectors to restore the functional deficits in muscle tissue itself have been reported to date. In this proposal, we will use delta-SG as the target disease gene, the delta-SG deficient hamster as the LGDM animal model, and rAAV as the gene delivery vector to test our general hypothesis that safe, efficient and sustained functional rescue of muscle deficiency can be achieved by genetic complementation of inherited muscular dystrophies with rAAV vectors. Specifically, we would like to achieve the following aims: 1) To study gene transfer efficiency and functional rescue in the LGMD hamster model by local intramuscular delivery of delta-SG-rAAV vectors and examine their short term ability to correlate the genetic defect in both skeletal and cardiac muscle. 2) To evaluate the gene therapy efficacy after systemic delivery of rAAV vectors through intra-artery or intra-ventricle injection. 3) To investigate the molecular kinetics and fate of rAAV vectors, especially after systemic vector delivery. 4) To develop new generation, high titer, helper-virus-free rAAV producer cells, which not only harbor vector and packaging genes, but also contain the necessary helper genes from adenovirus in a highly regulated and inducible manner.

National Institute of Health (NIH)
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
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Medical Biochemistry Study Section (MEDB)
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Lymn, Richard W
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University of Pittsburgh
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Yang, Lin; Xiao, Xiao (2013) Creation of a cardiotropic adeno-associated virus: the story of viral directed evolution. Virol J 10:50
He, Bo; Tang, Ru-hang; Weisleder, Noah et al. (2012) Enhancing muscle membrane repair by gene delivery of MG53 ameliorates muscular dystrophy and heart failure in ?-Sarcoglycan-deficient hamsters. Mol Ther 20:727-35
Qiao, Chunping; Yuan, Zhenhua; Li, Jianbin et al. (2012) Single tyrosine mutation in AAV8 and AAV9 capsids is insufficient to enhance gene delivery to skeletal muscle and heart. Hum Gene Ther Methods 23:29-37
Yang, Lin; Li, Juan; Xiao, Xiao (2011) Directed evolution of adeno-associated virus (AAV) as vector for muscle gene therapy. Methods Mol Biol 709:127-39
Hoshijima, Masahiko; Hayashi, Takeharu; Jeon, Young E et al. (2011) Delta-sarcoglycan gene therapy halts progression of cardiac dysfunction, improves respiratory failure, and prolongs life in myopathic hamsters. Circ Heart Fail 4:89-97
Yuan, Zhenhua; Qiao, Chunping; Hu, Peiqi et al. (2011) A versatile adeno-associated virus vector producer cell line method for scalable vector production of different serotypes. Hum Gene Ther 22:613-24
Qiao, C; Yuan, Z; Li, J et al. (2011) Liver-specific microRNA-122 target sequences incorporated in AAV vectors efficiently inhibits transgene expression in the liver. Gene Ther 18:403-10
Koppanati, B M; Li, J; Reay, D P et al. (2010) Improvement of the mdx mouse dystrophic phenotype by systemic in utero AAV8 delivery of a minidystrophin gene. Gene Ther 17:1355-62
Qiao, Chunping; Zhang, Wei; Yuan, Zhenhua et al. (2010) Adeno-associated virus serotype 6 capsid tyrosine-to-phenylalanine mutations improve gene transfer to skeletal muscle. Hum Gene Ther 21:1343-8
Powell, Frank L; Kim, B Cindy; Johnson, S Randall et al. (2009) Oxygen sensing in the brain--invited article. Adv Exp Med Biol 648:369-76

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