Antisense pre-treatment increases gene therapy efficacy in dystrophic muscles

Hum Mol Genet. 2016 Aug 15;25(16):3555-3563. doi: 10.1093/hmg/ddw201. Epub 2016 Jul 4.

Abstract

In preclinical models for Duchenne muscular dystrophy, dystrophin restoration during adeno-associated virus (AAV)-U7-mediated exon-skipping therapy was shown to decrease drastically after six months in treated muscles. This decline in efficacy is strongly correlated with the loss of the therapeutic AAV genomes, probably due to alterations of the dystrophic myofiber membranes. To improve the membrane integrity of the dystrophic myofibers at the time of AAV-U7 injection, mdx muscles were pre-treated with a single dose of the peptide-phosphorodiamidate morpholino (PPMO) antisense oligonucleotides that induced temporary dystrophin expression at the sarcolemma. The PPMO pre-treatment allowed efficient maintenance of AAV genomes in mdx muscles and enhanced the AAV-U7 therapy effect with a ten-fold increase of the protein level after 6 months. PPMO pre-treatment was also beneficial to AAV-mediated gene therapy with transfer of micro-dystrophin cDNA into muscles. Therefore, avoiding vector genome loss after AAV injection by PPMO pre-treatment would allow efficient long-term restoration of dystrophin and the use of lower and thus safer vector doses for Duchenne patients.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Dependovirus / genetics
  • Dystrophin / genetics*
  • Exons / genetics
  • Gene Transfer Techniques
  • Genetic Therapy*
  • Genetic Vectors / administration & dosage
  • Humans
  • Mice, Inbred mdx
  • Morpholinos / administration & dosage*
  • Muscle, Skeletal / drug effects
  • Muscle, Skeletal / pathology
  • Muscular Dystrophy, Animal / genetics
  • Muscular Dystrophy, Animal / therapy*
  • Muscular Dystrophy, Duchenne / genetics
  • Muscular Dystrophy, Duchenne / therapy*
  • Oligonucleotides, Antisense / administration & dosage*
  • Sarcolemma / drug effects
  • Sarcolemma / pathology

Substances

  • Dystrophin
  • Morpholinos
  • Oligonucleotides, Antisense