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Exon skipping restores dystrophin expression, but fails to prevent disease progression in later stage dystrophic dko mice

Gene Therapy volume 21, pages 785–793 (2014)Cite this article

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Abstract

Antisense therapy with both chemistries of phosphorodiamidate morpholino oligomers (PMOs) and 2′-O-methyl phosphorothioate has demonstrated the capability to induce dystrophin expression in Duchenne muscular dystrophy (DMD) patients in phase II-III clinical trials with benefit in muscle functions. However, potential of the therapy for DMD at different stages of the disease progression is not understood. In this study, we examined the effect of peptide-conjugated PMO (PPMO)-mediated exon skipping on disease progression of utrophin-dystrophin-deficient mice (dko) of four age groups (21–29, 30–39, 40–49 and 50+ days), representing diseases from early stage to advanced stage with severe kyphosis. Biweekly intravenous (i.v.) administration of the PPMO restored the dystrophin expression in nearly 100% skeletal muscle fibers in all age groups. This was associated with the restoration of dystrophin-associated proteins including functional glycosylated dystroglycan and neuronal nitric synthase. However, therapeutic outcomes clearly depended on severity of the disease at the time the treatment started. The PPMO treatment alleviated the disease pathology and significantly prolonged the life span of the mice receiving treatment at younger age with mild phenotype. However, restoration of high levels of dystrophin expression failed to prevent disease progression to the mice receiving treatment when disease was already at advanced stage. The results could be critical for design of clinical trials with antisense therapy to DMD.

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Acknowledgements

This work was supported by the Carolinas Muscular Dystrophy Research Endowment at the Carolinas HealthCare Foundation and Carolinas Medical Center, Charlotte, NC, Foundation to Eradicate Duchenne, Department of Defense (W81XWH-09-1-0599), and NIH/NICHD (U54 HD 071601-02). We thank AVI Biopharma (now Sarepta Therapeutics) for the supply of PPMOE23 for this study and Dr Mark Grady (Washington University, St Louis) and Dr Joshua Sanes (Harvard University, Cambridge) for providing mdx female mice heterozygous for utrophin (mdx;utr+/−).

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Authors and Affiliations

  1. Department of Neurology, McColl-Lockwood Laboratory for Muscular Dystrophy Research, Neuromuscular/ALS Center, Carolinas Medical Center, Charlotte, NC, USA

    B Wu, C Cloer, P Lu, S Milazi, M Shaban, S N Shah, L Marston-Poe & Q L Lu

  2. Department of Biomedical Sciences, College of Veterinary Medicine, Oregon State University, Corvallis, OR, USA

    H M Moulton

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  1. B Wu
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  2. C Cloer
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Correspondence to B Wu or Q L Lu.

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Wu, B., Cloer, C., Lu, P. et al. Exon skipping restores dystrophin expression, but fails to prevent disease progression in later stage dystrophic dko mice. Gene Ther 21, 785–793 (2014). https://doi.org/10.1038/gt.2014.53

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