EFFICIENT COEXPRESSION AND SECRETION OF ANTIATHEROGENIC HUMAN APOLIPOPROTEIN AL AND LECITHIN-CHOLESTEROL ACYLTRANSFERASE BY CULTURED MUSCLE-CELLS USING ADENOASSOCIATED VIRUS PLASMID VECTORS

Plasma apolipoprotein Al (apoAl) and lecithin-cholesterol acyltransfer ase (LCAT) play important roles in reverse cholesterol transport, prom oting the removal of excess cholesterol from peripheral cells and redu cing formation of atherosclerotic lesions. Gene augmentation of either apoAl or LCAT, or both, are thus attractive targets for prevention or treatment of atherosclerosis. With the eventual aim of safe and effic ient gene delivery to skeletal muscle, our chosen secretory platform f or systemic delivery of antiatherogenic proteins, we have constructed conventional and AAV-based plasmid vectors containing human apoAl or L CAT cDNAs; their efficacy was tested by lipoplex transfection of mouse C2C12 muscle cells or human 293 cells. The secretion of apoAl or LCAT by transduced cultures was two- to five-fold higher using AA V-based plasmid vectors than conventional plasmid vectors. Additionally, cells transfected with a bicistronic AA V-based vector containing an intern al ribosome entry site (IRES) efficiently expressed both apoAl and LCA T simultaneously. Furthermore, AAV-based vector sequences were retaine d by host cells, whereas those of conventional plasmid vectors were lo st. These studies indicate that ectopic overexpression of apoAl and LC AT in muscle tissue using AAV-based plasmid vectors might provide a fe asible anti-atherogenic strategy in vivo.