Induced dystrophin exon skipping in human muscle explants

Antisense oligonucleotide (AO) manipulation of pre-mRNA splicing of the dystrophin gene shows potential in overcoming Duchenne muscular dystrophy (DMD)-causing mutations. AOs are used to alter dystrophin pre-mRNA splicing to reduce the consequences of nonsense or frame-shifting mutations that would otherwise prematurely terminate translation. We have developed a panel of AOs, targeting 77 out of 79 exons, to address mutations across the human dystrophin gene transcript that will restore the reading frame by either removing single or multiple exons. The aim of this study was to demonstrate that AOs could be used to induce exon skipping in human muscle. To date, this approach has been limited to studies using animal models or cultured monolayers of human muscle cells. We used different AO chemistries to induce exon skipping in human muscle explants, derived from normal and DMD human tissue, with great success. We propose that inducing exon skipping in human muscle explants is closer to in vivo conditions than cells in monolayer cultures, and that it is possible that explant studies may be used as a further tool to assist in the design of the AOs and evolution of different AO chemistries