Abstract 628: Overexpression of ABCA1 in Cultured Endothelial Cells Using Helper-Dependent Adenovirus Enhances ApoAI-Mediated Cholesterol Efflux and has Anti-Inflammatory Effects

Background: ABCA1 removes cholesterol from vascular wall cells via apoAI-mediated efflux, generating HDL that transports cholesterol to the liver for excretion. This process of reverse cholesterol transport is atheroprotective; therefore, strategies that increase vascular wall ABCA1 may prevent or reverse atherosclerosis. Cholesterol efflux mediated by ABCA1 and apoAI can also have anti-inflammatory effects; however, excess depletion of cellular cholesterol can cause cell stress and apoptosis. We tested whether transducing endothelial cells (EC) with a helper-dependent adenoviral vector that expresses ABCA1 (HDAdABCA1) enhances apoAI-mediated cholesterol efflux and reduces inflammatory markers without causing cellular toxicity. Methods: We cloned rabbit ABCA1, constructed HDAdABCA1, transduced bovine aortic EC (BAEC) with either HDAdABCA1 or empty vector (HDAdNull). We measured ABCA1 protein by immunoblotting and apoAI-mediated cholesterol efflux by loading EC with 3 [H] cholesterol, then adding apoAI protein to serum-free medium. We assessed EC phenotype using MTT (metabolic activity), BrdU (proliferation), wound-healing assay (migration), and flow cytometry (apoptosis). We measured ICAM-1, VCAM-1, IL-6, and TNFα mRNA in transduced EC by qRT-PCR both under basal conditions and after serum-starvation, addition of apoAI, and LPS challenge. Results: We observed a ~3-fold increase in ABCA1 protein in EC transduced with HDAdABCA1 and a ~2-fold increase in apoAI-mediated cholesterol efflux (P Conclusions: HDAdABCA1 increases EC ABCA1 expression and enhances apoAI-mediated cholesterol efflux, but does not cause toxicity or increase inflammatory markers. In contrast, ABCA1 overexpression appears to have anti-inflammatory effects. Future studies will test if HDAdABCA1 decreases lipid rafts, and whether overexpression of ABCA1 in EC in vivo is atheroprotective.