Inhibition of adhesion molecule expression on human venous endothelial cells by non-viral siRNA transfection

Objective: Expression of adhesion molecule receptors on venous endothelial cells crucially influences the fate of venous grafts by mediating leukocyte-endothelium interactions. These interactions include adhesion of leukocytes to the endothelium, followed by transendothelial migration, leading to neointimal hyperplasia and finally graft occlusion. Therefore, inhibition of adhesion molecule expression may be a promising strategy to improve the quality of venous grafts. We tested the efficiency of non-viral transfection of human venous endothelial cells with short interfering RNA (siRNA) to specificially down-regulate adhesion molecule expression. Methods: Primary cultures of human venous endothelial cells (HVEC) were examined for expression of the adhesion molecules ICAM1, VCAM1 and E-selectin (SELE) after non viral siRNA transfection. Adhesion molecule expression was measured by flow cytometry, real-time PCR and immunoblotting after stimulation with TNF-alpha, an inflammatory cytokine. Results: Non-transfected cells showed a strong increase of adhesion molecule expression following cytokine stimulation (p<0.01). Upon transfection with specific siRNAs a six fold decrease in ICAM1 (p<0.001) and SELE expression and cell positivity (p<0.05) and a twofold decrease in VCAM1 expression and cell positivity (p<0.01) could be observed. SiRNA-mediated gene suppression of adhesion molecules was also reflected by corresponding decreases in adhesion protein and transcript levels. Conclusions: The expression of adhesion molecules on HVECs can be effectively inhibited by specific siRNAs using a safe, non-viral transfection approach. This is a promising tool to precondition venous bypass grafts in order to interfere with endothelium-leukocyte interactions and to prohibit neointima thickening or atherosclerosis, which are regarded to be the most important causes of venous graft failure.