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A canine minidystrophin is functional and therapeutic in mdx mice

Gene Therapy volume 15, pages 1099–1106 (2008)Cite this article

Abstract

Duchenne muscular dystrophy (DMD) is the most common and lethal genetic muscle disorder lacking a curative treatment. We wish to use the dystrophin-deficient golden retriever muscular dystrophy (GRMD) dog, a canine model of DMD, to investigate adeno-associated virus (AAV) vector-mediated minidystrophin gene therapy. The dog model is useful in evaluating vector dose requirement and immunological consequences owing to its large size and outbred nature. In this study, we have cloned and constructed a canine minidystrophin gene vector. Owing to limited availability of the GRMD dogs, here we first examined the functions and therapeutic effects of the canine minidystrophin in the mdx mouse model. We observed efficient minigene expression without cellular immune responses in mdx mice after AAV1-cMinidys vector intramuscular injection. We also observed restoration of the missing dystrophin-associated protein complex (DPC) onto the sarcolemma, including sarcoglycans and dystrobrevin, and a partial restoration of α-syntrophin and neural nitric oxide synthase (nNOS). In addition, minidystrophin treatment ameliorated dystrophic pathology, such as fibrosis and myofiber central nucleation (CN). CN remained minimal (<2%) after AAV injection in the neonatal mdx mice and was reduced from more than 75% to about 25% after AAV injection in adult mdx mice. Finally, in vivo cell membrane leakage test with Evans blue dye showed that the canine minidystrophin could effectively protect the myofiber plasma membrane integrity. Our results, thus, demonstrated the functionality and therapeutic potential of the canine minidystrophin and paved its way for further testing in the GRMD dog model.

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Acknowledgements

This study is supported by NIH grants Muscular Dystrophy Center for Research and Cooperation (MDCRC) AR50733 and AR50595. We thank Mr Mengnan Tian for technical assistance and Dr Tong Zhu for advice and discussion.

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Authors and Affiliations

  1. Department of Orthopedic Surgery, University of Pittsburgh, Pittsburgh, PA, USA

    B Wang & L Zhou

  2. Division of Molecular Pharmaceutics, University of North Carolina School of Pharmacy, Chapel Hill, NC, USA

    J Li, C Qiao, C Chen, P Hu & X Xiao

  3. Cardiovascular Institute, University of Pittsburgh, Pittsburgh, PA, USA

    X Zhu

  4. Department of Pathology, University of North Carolina School of Medicine, Chapel Hill, NC, USA

    J Bogan & J Kornegay

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  1. B Wang
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  2. J Li
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  3. C Qiao
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  4. C Chen
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  5. P Hu
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  6. X Zhu
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  7. L Zhou
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  10. X Xiao
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Correspondence to X Xiao.

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Wang, B., Li, J., Qiao, C. et al. A canine minidystrophin is functional and therapeutic in mdx mice. Gene Ther 15, 1099–1106 (2008). https://doi.org/10.1038/gt.2008.70

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