title: Exon skipping and gene transfer restore dystrophin expression in hiPSC-cardiomyocytes harbouring DMD mutations.
abstract: With an incidence of ~1:3,500 to 5,000 in male children, Duchenne muscular dystrophy (DMD) is an X-linked disorder in which progressive muscle degeneration occurs and affected boys usually die in their twenties or thirties. Cardiac involvement occurs in 90% of patients and heart failure accounts for up to 40% of deaths. To enable new therapeutics such as gene therapy and exon skipping to be tested in human cardiomyocytes, we produced human induced pluripotent stem cells (hiPSC) from seven patients harbouring mutations across the DMD gene. Mutations were retained during differentiation and analysis indicated the cardiomyocytes showed a dystrophic gene expression profile. Antisense oligonucleotide-mediated skipping of exon 51 restored dystrophin expression to ~30% of normal levels in hiPSC-cardiomyocytes carrying exon 47-50 or 48-50 deletions. Alternatively, delivery of a dystrophin minigene to cardiomyocytes with a deletion in exon 35 or a point mutation in exon 70 allowed expression levels similar to those seen in healthy cells. This demonstrates that DMD hiPSC-cardiomyocytes provide a novel tool to evaluate whether new therapeutics can restore dystrophin expression in the heart.
authors.forename: Emily
authors.initials: E
authors.lastname: Dick
authors.forename: Spandan
authors.initials: S
authors.lastname: Kalra
authors.forename: David
authors.initials: D
authors.lastname: Anderson
authors.forename: Vinoj
authors.initials: V
authors.lastname: George
authors.forename: Morten
authors.initials: M
authors.lastname: Ritson
authors.forename: Steve
authors.initials: S
authors.lastname: Laval
authors.forename: Rita
authors.initials: R
authors.lastname: Barresi
authors.forename: Annemieke
authors.initials: A
authors.lastname: Aartsma-Rus
authors.forename: Hanns
authors.initials: H
authors.lastname: Lochmuller
authors.forename: Chris
authors.initials: C
authors.lastname: Denning