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Systemic delivery of genes to striated muscles using adeno-associated viral vectors

Nature Medicine volume 10, pages 828–834 (2004)Cite this article

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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Figure 1: Muscle transduction (blue) following systemic rAAV6 administration.
Figure 2: β-Gal activity in muscles after systemic rAAV6 administration.
Figure 3: β-Gal activity and vector genome copy numbers in muscles and organs after systemic administration of rAAV6 vectors with VEGF.
Figure 4: Myocardial transduction (blue) in adult mice 11 days after systemic administration of 1 × 1012 vector genomes of rAAV6–CMV–lacZ and 10 μg VEGF.
Figure 5: Systemic delivery of microdystrophin to dystrophic mice.
Figure 6: Sustained expression of a therapeutic human microdystrophin protein is achieved after intravenous administration of rAAV6 vectors, without significant cellular infiltration.

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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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Author notes

  1. Paul Gregorevic and Michael J Blankinship: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Neurology, Senator Paul D. Wellstone Muscular Dystrophy Cooperative Research Center, The University of Washington, Seattle, 98195, Washington, USA

    Paul Gregorevic, Michael J Blankinship, Robert W Crawford, Leonard Meuse & Jeffrey S Chamberlain

  2. Department of Medicine, Senator Paul D. Wellstone Muscular Dystrophy Cooperative Research Center, The University of Washington, Seattle, 98195, Washington, USA

    James M Allen, Daniel G Miller, David W Russell & Jeffrey S Chamberlain

  3. Department of Biochemistry, Senator Paul D. Wellstone Muscular Dystrophy Cooperative Research Center, The University of Washington, Seattle, 98195, Washington, USA

    David W Russell & Jeffrey S Chamberlain

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  1. Paul Gregorevic
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Correspondence to Jeffrey S Chamberlain.

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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.

Supplementary information

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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