Abstract
Muscular dystrophies are a genetically and phenotypically heterogeneous group of degenerative muscle diseases. A subset of them are due to genetic deficiencies in proteins which form the dystrophin-associated complex at the membrane of the myofibers. In this report, we utilized recombinant adeno-associated virus containing a U7 cassette carrying an antisense sequence aimed at inducing exon skipping of the dystrophin gene or containing the α-sarcoglycan gene to alleviate the dystrophic phenotype of the mdx and Sgca-null mice, respectively. As these diseases are characterized by cycle of degeneration/regeneration, we postulated that a reporter gene coadministered at the time of the treatment would make it possible to follow the extent of muscle repair. We observed that the murine secreted alkaline phosphatase (muSeAP) level was very much lower in these animal models than in normal mice. Upon treatment of the dystrophic muscle by gene transfer, the level of muSeAP was restored and correlated with the expression of the therapeutic transgene and with the level of muscle improvement. The system described here provides a simple and noninvasive procedure for monitoring the outcome of a therapeutic strategy involving cell survival.
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Acknowledgements
We acknowledge the excellent expertise of the In vivo department of Généthon, especially Ludovic Arandel for animal procedures and Daniel Stockholm for help in the real-time quantitative RT-PCR experiments. We are grateful to Susan Cure for critical reading of the manuscript. We would like to thank the Howard Hughes Medical Institute (Iowa City, USA) for providing us with Sgca-null mice and Vincent Thuillier (Gencell, Vitry-sur-Seine, France) for the pVT20 plasmid. AG is the recipient of a fellowship from the Ministère de l'Éducation Nationale, de la Recherche et de la Technologie. This work was funded by the Association Française contre les Myopathies, Genopole® (Evry) and the Fondation pour la Recherche Médicale.
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Bartoli, M., Poupiot, J., Goyenvalle, A. et al. Noninvasive monitoring of therapeutic gene transfer in animal models of muscular dystrophies. Gene Ther 13, 20–28 (2006). https://doi.org/10.1038/sj.gt.3302594
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DOI: https://doi.org/10.1038/sj.gt.3302594
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