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

Abstract

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.

• Authors: Sandra T Cooper, Eddy Kizana, Jonathon D Yates, Harriet P Lo, Nan Yang, Zhan He Wu, Ian E Alexander, Kathryn N North
• Journal: Neuromuscular disorders : NMD (Neuromuscul Disord) Vol. 17 Issue 4 Pg. 276-84 (Apr 2007) ISSN: 0960-8966 [Print] England
• PMID: 17303423 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Dystrophin
• MyoD Protein
• RNA, Messenger
• Actinin

Topics

• 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, metabolism)
• Protein Deficiency (metabolism)
• RNA, Messenger
• Reverse Transcriptase Polymerase Chain Reaction (methods)
• Time Factors
• Transduction, Genetic (methods)