Dystrophinopathy carrier determination and detection of protein deficiencies in muscular dystrophy using lentiviral MyoD-forced myogenesis

Neuromuscul Disord. 2007 Apr;17(4):276-84. doi: 10.1016/j.nmd.2006.12.010. Epub 2007 Feb 15.


The objective of this study is to expand the applications of MyoD-forced myogenesis for research and diagnosis of human muscle disorders using a lentiviral vector (LVhMyoD) for efficient trans-differentiation of patient primary cells. LVhMyoD transduced cells readily formed striated, multinucleate myotubes expressing a wide range of genes associated with muscular dystrophy (dystrophin, dysferlin, sarcoglycans, caveolin-3) and congenital myopathy (nebulin, actin, desmin, tropomyosin, troponin). We demonstrate that MyoD gene-modified fibroblasts reproduce protein deficiencies associated with different forms of muscular dystrophy, and confirm that LVhMyoD gene-modified chorionic villus can be used successfully to determine the dystrophin status of the developing fetus, augmenting prenatal diagnosis of dystrophinopathy patients. Using muscle-specific cDNA derived from LVhMyoD gene-modified patient cells, we identified a female carrier bearing a large dystrophin deletion and a previously unidentified non-coding splice-site mutation within dystrophin in a Becker muscular dystrophy patient. This study highlights the significant potential of lentiviral MyoD-forced myogenesis for study of a wide range of human muscle disorders; a field constrained by the limited availability of human tissue. LVhMyoD gene-modified patient cells provide a renewable source of mutant protein and muscle-specific mRNA, facilitating accelerated mutation screening of large genes, molecular analyses of splicing abnormalities and study of disease-causing mutations.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actinin / metabolism
  • Cell Differentiation
  • Cells, Cultured
  • Chorionic Villi / pathology
  • DNA Mutational Analysis
  • Dystrophin / metabolism*
  • Female
  • Fibroblasts / pathology
  • Humans
  • Lentivirus / physiology*
  • Male
  • Muscle Development / physiology
  • Muscular Dystrophies / pathology*
  • Mutation
  • MyoD Protein / genetics
  • MyoD Protein / metabolism*
  • Protein Deficiency / metabolism*
  • RNA, Messenger
  • Reverse Transcriptase Polymerase Chain Reaction / methods
  • Time Factors
  • Transduction, Genetic / methods


  • Dystrophin
  • MyoD Protein
  • RNA, Messenger
  • Actinin