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Restoration of all dystrophin protein interactions by functional domains in trans does not rescue dystrophy

Gene Therapy volume 13, pages 744–751 (2006)Cite this article

Abstract

Rescue of dystrophic skeletal muscle in mdx and utrophin/dystrophin-deficient (dko) mouse models by reintroduction of dystrophin has validated gene therapy as a potential therapeutic approach for Duchenne muscular dystrophy. However, the size of the dystrophin gene exceeds the capacity of adeno-associated viral (AAV) vectors. Dystrophin provides a mechanical link at the muscle membrane by direct binding of its amino-terminal and cysteine-rich domains to actin and a transmembrane protein complex, respectively. It has not been investigated whether restoration of these two tethering functions by two separate dystrophin molecules is sufficient to prevent dystrophic pathologies. We examine the effect of coexpression of the amino-terminal and cysteine-rich domains from separate dystrophin transgenes, Δcys and Dp71, on the dystrophic phenotype. Expression of individual dystrophin domains from multiple vectors would effectively expand AAV capacity. Although both Δcys and Dp71 colocalize at the membrane, there is no improvement of dystrophic pathology. The fiber-type and neuromuscular junction abnormalities of dko mice that are ameliorated by the Δcys transgene are not further improved or disrupted by Dp71. Separate truncated dystrophins, which together restore all protein interactions and scaffolding for signaling molecules, are not sufficient to ameliorate the dystrophic phenotype and therefore dystrophin domains in trans cannot be used to increase the effective cloning capacity for AAV-mediated gene therapy.

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Acknowledgements

This work was supported by the Muscular Dystrophy Association and a Career Award in the Biomedical Sciences from the Burroughs Wellcome Fund (to JRF). KLG was supported by a Chemistry/Biology Interface training grant from NIH (T32 GM08512).

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Authors and Affiliations

  1. Department of Molecular and Cellular Biochemistry, College of Medicine, The Ohio State University, Columbus, OH, USA

    K L Gardner, J A Kearney, J D Edwards & J A Rafael-Fortney

  2. Department of Chemistry, The Ohio State University, 100 Celeste Laboratory, Columbus, OH, USA

    K L Gardner

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  1. K L Gardner
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  2. J A Kearney
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  4. J A Rafael-Fortney
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Correspondence to J A Rafael-Fortney.

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Gardner, K., Kearney, J., Edwards, J. et al. Restoration of all dystrophin protein interactions by functional domains in trans does not rescue dystrophy. Gene Ther 13, 744–751 (2006). https://doi.org/10.1038/sj.gt.3302686

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