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

被引:29
作者
Cooper, Sandra T.
Kizana, Eddy
Yates, Jonathon D.
Lo, Harriet P.
Yang, Nan
Wu, Zhan He
Alexander, Ian E.
North, Kathryn N.
机构
[1] Childrens Hosp Westmead, Neurogenet Res Unit, Sydney, NSW 2145, Australia
[2] Childrens Hosp Westmead, Inst Neuromuscular Res, Sydney, NSW 2145, Australia
[3] Univ Sydney, Discipline Paediat & Child Hlth, Sydney, NSW 2006, Australia
[4] Childrens Hosp Westmead, Gene Therapy Res Unit, Sydney, NSW, Australia
[5] Childrens Med Res Inst, Sydney, NSW, Australia
[6] Childrens Hosp Westmead, Dept Cytogenet, Sydney, NSW, Australia
基金
英国医学研究理事会;
关键词
MyoD-forced myogenesis; muscular dystrophy; myopathy; diagnosis;
D O I
10.1016/j.nmd.2006.12.010
中图分类号
R74 [神经病学与精神病学];
学科分类号
摘要
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. (c) 2006 Elsevier B.V. All rights reserved.
引用
收藏
页码:276 / 284
页数:9
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