Abstract
A major obstacle limiting gene therapy for diseases of the heart and skeletal muscles is an inability to deliver genes systemically to muscles of an adult organism. Systemic gene transfer to striated muscles is hampered by the vascular endothelium, which represents a barrier to distribution of vectors via the circulation. Here we show the first evidence of widespread transduction of both cardiac and skeletal muscles in an adult mammal, after a single intravenous administration of recombinant adeno-associated virus pseudotype 6 vectors. The inclusion of vascular endothelium growth factor/vascular permeability factor, to achieve acute permeabilization of the peripheral microvasculature, enhanced tissue transduction at lower vector doses. This technique enabled widespread muscle-specific expression of a functional micro-dystrophin in the skeletal muscles of dystrophin-deficient mdx mice, which model Duchenne muscular dystrophy. We propose that these methods may be applicable for systemic delivery of a wide variety of genes to the striated muscles of adult mammals.
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Acknowledgements
We thank S.D. Hauschka for helpful discussions; D. Duan for the AAV MCS vector plasmids pDD2 and pDD344; J. Kleinschmidt for the packaging plasmid pDG; A.D. Miller for the packaging cell line 293; J. Han for the rAAV6–CK6–lacZ construct; and M. Haraguchi and S. Oakley for technical assistance. This work was supported by grants from the National Institutes of Health and the Muscular Dystrophy Association (USA) to J.S.C.
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The University of Washington, which employs the authors, has filed for a patent application relating to some of the results reported in this manuscript.
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Supplementary Fig. 1
Intravenous administration of rAAV6 vectors and VEGF is not associated with evidence of acute organ toxicity. (PDF 389 kb)
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Gregorevic, P., Blankinship, M., Allen, J. et al. Systemic delivery of genes to striated muscles using adeno-associated viral vectors. Nat Med 10, 828–834 (2004). https://doi.org/10.1038/nm1085
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DOI: https://doi.org/10.1038/nm1085
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