Experimental model of gene transfection in healthy canine myocardium: perspectives of gene therapy for ischemic heart disease.
OBJECTIVE
To assess the transfection of the gene that encodes green fluorescent protein (GFP) through direct intramyocardial injection.


METHODS
The pREGFP plasmid vector was used. The EGFP gene was inserted downstream from the constitutive promoter of the Rous sarcoma virus. Five male dogs were used (mean weight 13.5 kg), in which 0.5 mL of saline solution (n=1) or 0.5 mL of plasmid solution containing 0.5 micro g of pREGFP/dog (n=4) were injected into the myocardium of the left ventricular lateral wall. The dogs were euthanized 1 week later, and cardiac biopsies were obtained.


RESULTS
Fluorescence microscopy showed differences between the cells transfected and not transfected with pREGFP plasmid. Mild fluorescence was observed in the cardiac fibers that received saline solution; however, the myocardial cells transfected with pREGFP had overt EGFP expression.


CONCLUSIONS
Transfection with the EGFP gene in healthy canine myocardium was effective. The reproduction of this efficacy using vascular endothelial growth factor (VEGF) instead of EGFP aims at developing gene therapy for ischemic heart disease.
Kalil
Renato A K
RA
Instituto de Cardiologia do Rio Grande do Sul, Fundação Universitária de Cardiologia, Porto Alegre, RS. pesquisa@cardnet.tche.br
Teixeira
Leonardo A K
LA
Mastalir
Eduardo T
ET
Moreno
Paulo
P
Fricke
Cecília H
CH
Nardi
Nance B
NB
D000818:Animals; D004285:Dogs; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000627:therapeutic use; D049452:Green Fluorescent Proteins; D007202:Indicators and Reagents; D008164:Luminescent Proteins / Q000235:genetics*; D008297:Male; D008957:Models, Genetic; D017202:Myocardial Ischemia / Q000628:therapy*; D009206:Myocardium; D014162:Transfection
D007202:Indicators and Reagents; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins