Exon 45 skipping through U1-snRNA antisense molecules recovers the Dys-nNOS pathway and muscle differentiation in human DMD myoblasts

Valentina Cazzella, Julie Martone, Chiara Pinnarò, Tiziana Santini, Shyam Sundar Twayana, Olga Sthandier, Adele D'amico, Valeria Ricotti, Enrico Bertini, Francesco Muntoni, Irene Bozzoni

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Abstract

Exon skipping has been demonstrated to be a successful strategy for the gene therapy of Duchenne muscular dystrophy (DMD): The rational being to convert severe Duchenne forms into milder Becker ones. Here, we show the selection of U1 snRNA-antisense constructs able to confer effective rescue of dystrophin synthesis in a Δ44 Duchenne genetic background, through skipping of exon 45; moreover, we demonstrate that the resulting dystrophin is able to recover timing of myogenic marker expression, to relocalize neuronal nitric oxide synthase (nNOS) and to rescue expression of miRNAs previously shown to be sensitive to the Dystrophin-nNOS-HDAC2 pathway. Becker mutations display different phenotypes, likely depending on whether the shorter protein is able to reconstitute the wide range of wild-type functions. Among them, efficient assembly of the dystrophin-associated protein complex (DAPC) and nNOS localization are important. Comparing different Becker deletions we demonstrate the correlation between the ability of the mutant dystrophin to relocalize nNOS and the expression levels of two miRNAs, miR-1 and miR29c, known to be involved in muscle homeostasis and to be controlled by the Dys-nNOS-HDAC2 pathway.

Original languageEnglish
Pages (from-to)2134-2142
Number of pages9
JournalMolecular Therapy
Volume20
Issue number11
DOIs
Publication statusPublished - Nov 2012

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ASJC Scopus subject areas

  • Molecular Biology
  • Molecular Medicine
  • Genetics
  • Drug Discovery
  • Pharmacology

Cite this

Cazzella, V., Martone, J., Pinnarò, C., Santini, T., Twayana, S. S., Sthandier, O., D'amico, A., Ricotti, V., Bertini, E., Muntoni, F., & Bozzoni, I. (2012). Exon 45 skipping through U1-snRNA antisense molecules recovers the Dys-nNOS pathway and muscle differentiation in human DMD myoblasts. Molecular Therapy, 20(11), 2134-2142. https://doi.org/10.1038/mt.2012.178