title: Dual exon skipping in myostatin and dystrophin for Duchenne muscular dystrophy.
abstract: BACKGROUND
  Myostatin is a potent muscle growth inhibitor that belongs to the Transforming Growth Factor-β (TGF-β) family. Mutations leading to non functional myostatin have been associated with hypermuscularity in several organisms. By contrast, Duchenne muscular dystrophy (DMD) is characterized by a loss of muscle fibers and impaired regeneration. In this study, we aim to knockdown myostatin by means of exon skipping, a technique which has been successfully applied to reframe the genetic defect of dystrophin gene in DMD patients.
  
  
  METHODS
  We targeted myostatin exon 2 using antisense oligonucleotides (AON) in healthy and DMD-derived myotubes cultures. We assessed the exon skipping level, transcriptional expression of myostatin and its target genes, and combined myostatin and several dystrophin AONs. These AONs were also applied in the mdx mice models via intramuscular injections.
  
  
  RESULTS
  Myostatin AON induced exon 2 skipping in cell cultures and to a lower extent in the mdx mice. It was accompanied by decrease in myostatin mRNA and enhanced MYOG and MYF5 expression. Furthermore, combination of myostatin and dystrophin AONs induced simultaneous skipping of both genes.
  
  
  CONCLUSIONS
  We conclude that two AONs can be used to target two different genes, MSTN and DMD, in a straightforward manner. Targeting multiple ligands of TGF-beta family will be more promising as adjuvant therapies for DMD.
authors.affiliation: Center for Human and Clinical Genetics, Leiden University Medical Center, Postzone S4-P, PO Box 9600, Leiden, 2300RC, the Netherlands.
authors.forename: Dwi U
authors.initials: DU
authors.lastname: Kemaladewi
authors.forename: Willem M H
authors.initials: WM
authors.lastname: Hoogaars
authors.forename: Sandra H
authors.initials: SH
authors.lastname: van Heiningen
authors.forename: Samuel
authors.initials: S
authors.lastname: Terlouw
authors.forename: David J J
authors.initials: DJ
authors.lastname: de Gorter
authors.forename: Johan T
authors.initials: JT
authors.lastname: den Dunnen
authors.forename: Gert Jan B
authors.initials: GJ
authors.lastname: van Ommen
authors.forename: Annemieke
authors.initials: A
authors.lastname: Aartsma-Rus
authors.forename: Peter
authors.initials: P
authors.lastname: ten Dijke
authors.forename: Peter A C
authors.initials: PA
authors.lastname: 't Hoen
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D016189:Dystrophin / Q000235:genetics*; D005091:Exons / Q000235:genetics*; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D051379:Mice; D018101:Mice, Inbred mdx; D024510:Muscle Development / Q000187:drug effects / Q000235:genetics; D018485:Muscle Fibers, Skeletal / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D009137:Muscular Dystrophy, Animal / Q000235:genetics; D020388:Muscular Dystrophy, Duchenne / Q000235:genetics*; D032446:Myoblasts / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D055435:Myostatin / Q000235:genetics* / Q000378:metabolism; D016376:Oligonucleotides, Antisense / Q000008:administration & dosage / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism
chemical_list: D016189:Dystrophin; D055435:Myostatin; D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger