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Restoration of dystrophin expression in mdx mice by intravascular injection of naked DNA containing full-length dystrophin cDNA

Gene Therapy volume 11, pages 901–908 (2004)Cite this article

Abstract

Duchenne muscular dystrophy (DMD) is a lethal, X-linked, recessive disease caused by a defect in the dystrophin gene. No effective therapy is available. Dystrophin gene transfer to skeletal muscle has been proposed as a treatment for DMD. However, successful treatment for DMD requires restoration of dystrophin in the affected muscle fibers to at least 20% of the normal level. Current gene transfer methods such as intramuscular injection of viral vector or naked DNA can only transfect a small area of muscle, and therefore is of little clinical utility. We have developed a semisystemic method for gene transfer into skeletal muscle of mdx mice, an animal model for DMD. Naked DNA was injected through the tail artery or vein of mice, in which the aorta and the vena cava were clamped at the location just below the kidneys. The DNA solution was thus forced into the blood vessels of both legs. Luciferase gene expression was detected in all muscle groups in both legs. The effects of injection speed, injection volume, and ischemia time on gene expression were also optimized. LacZ staining was used to check the spread of gene expression in muscle. Although the percentage of transfected fibers was modest (∼10%), β-galactosidase was found in all muscle groups of both legs. Finally, plasmid DNA encoding full-length dystrophin gene was injected into mdx mice and widespread restoration of dystrophin protein was observed in all muscles of both hind limbs. In conclusion, these results demonstrate that the semisystemic delivery of naked DNA is a potential approach towards the long-term goal of gene therapy for DMD.

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Acknowledgements

This work was supported in part by Muscular Dystrophy Association grant and NIH grant PO1 AR45925 (L. Huang) and grant from the Uehara memeriol Foundation (M. Nishikawa). We thank Dr Paula Clemens for providing the dystrophin plasmid and mdx mice. Kenneth W Liang and Makiya Nishikawa were postdoctoral fellows of Duchenne Muscular Dystrophy Research Center, University of Pittsburgh.

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Authors and Affiliations

  1. Center for Pharmacogenetics, School of Pharmacy, University of Pittsburgh, Pittsburgh, PA, USA

    K W Liang, M Nishikawa, F Liu, B Sun & L Huang

  2. Department of Biological Sciences, Carnegie Mellon University, Pittsburgh, PA, USA

    Q Ye

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  1. K W Liang
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  2. M Nishikawa
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  6. L Huang
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Liang, K., Nishikawa, M., Liu, F. et al. Restoration of dystrophin expression in mdx mice by intravascular injection of naked DNA containing full-length dystrophin cDNA. Gene Ther 11, 901–908 (2004). https://doi.org/10.1038/sj.gt.3302239

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