Molecular cardiomyoplasty: potential cardiac gene therapy for chronic heart failure.
In this study, we evaluated the feasibility of converting cardiac fibroblasts into skeletal muscle cells by forced expression of the MyoD gene, one of the basic helix-loop-helix myogenic factors. Primary cardiac fibroblasts, isolated from newborn rats, were infected with retrovirus-carrying sense or antisense MyoD gene. Ten days after infection, expression of MyoD protein was demonstrated in 95% of cells infected with sense MyoD virus by intense nuclear immunostaining with a MyoD polyclonal antibody. In contrast, none of the cells infected with antisense MyoD virus showed staining. On withdrawal of serum, 95% of MyoD positive cells became elongated and, in the presence of appropriate cell density, fused to form multinucleated myotubes, morphologically similar to striated muscle cell. Expression of downstream myogenic differentiation markers, myosin heavy chain and myocyte-specific enhancer factor 2, in 95% of these myotubes were detected by intense cytoplasmic and nuclear immunostaining, respectively, with specific antibodies. In contrast, no detectable staining was noted in MyoD negative cells. Spontaneous contractile movements were noted in a few clusters of myotubes. In summary, cardiac fibroblasts were able to be converted into bonafide potentially functional skeletal myocytes as shown by definitive morphologic and biochemical changes. Further studies with in vivo models are needed to explore this unique molecular strategy to treat patients with chronic heart failure.
Tam
S K
SK
Cardiac Surgical Unit, Massachusetts General Hospital, Boston, USA.
Gu
W
W
Nadal-Ginard
B
B
D000818:Animals; D000831:Animals, Newborn; D002454:Cell Differentiation; D002908:Chronic Disease; D005347:Fibroblasts / Q000166:cytology*; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006333:Heart Failure / Q000628:therapy*; D017570:MyoD Protein / Q000235:genetics*; D009206:Myocardium / Q000166:cytology*; D051381:Rats; D017208:Rats, Wistar; D012190:Retroviridae / Q000235:genetics
D017570:MyoD Protein