Intramiocardial gene silencing of epidermal growth factor receptor (EGFR) by using lentiviral vectors. Functional evidence

BREA MS; DÍAZ RG; ESCUDERO DS; LEPORACE GUIMIL J; CALDIZ CI.; MORGAN PE; PÉREZ NG

Bariloche

Simposio; The Third South American Symposium in Signal Transduction and Molecular Medicine (SISTAM); 2015

American Society for Biochemistry and Molecular Biology

The aim of our work was the development of a 3rd generation lentivirus carrying a small interfering RNA (siRNA) capable to silence the EGFR in the myocardium, and being an experimental tool for the study of the possible rol of the receptor in the pathological cardiac hypertrophy. For virus production, HEK 293T cells were transfected with the packaging vectors and the silencing vector, and the culture medium was collected for 72 hours. A titer of 1x108 TU/mL was obtained. The lentivirus was injected into a rat cardiac left ventricular wall (n=5), and as control, a second group of sham rats were injected with phosphate buffer saline (PBS, n=5). Four-week later, EGFR protein expression in the left ventricle of siRNA-EGFR expressing rats showed 48 ± 15 % reduction compared to sham animals (100 ± 6 %, P