Abstract 396: Acute Hepatic ACAT2 Knockdown Transiently Increases Plasma and Hepatic Free Cholesterol and Fecal Neutral Sterol Excretion

Abstract

In spite of the advent of statins, atherosclerotic coronary vascular disease (ASCVD) remains the number one killer of Americans. A way to reduce LDL cholesterol, the primary risk factor of ASCVD, is to increase cholesterol excretion from the body. Our group and others have recently found that cholesterol excretion can be facilitated by both biliary and non-biliary pathways. The lipoprotein that delivers cholesterol from the liver through the plasma to the small intestine for transintestinal cholesterol excretion (TICE) is not yet known. We have previously shown that chronic knockdown in mice of hepatic acyl-CoA cholesterol acyltransferase 2 (ACAT2), a cellular enzyme that converts free cholesterol (FC) into cholesteryl ester (CE), appeared to cause the formation of hepatic apoB-containing lipoproteins that preferentially trafficked cholesterol to the small intestine for TICE. We tested the hypothesis that the plasma concentration of TICE-competent, apoB-containing lipoproteins, could be increased by preloading the liver with cholesterol and then acutely depleting the cholesterol by knocking down hepatic ACAT2 with antisense oligonucleotides (ASO). After feeding a high cholesterol (0.2% wt/wt) diet for six weeks, C57BL/6 mice were treated with control non-targeting ASO or ACAT2 ASO for one or two weeks. After only one week of ACAT2 knockdown (ACAT2KD) hepatic ACAT2 protein expression was decreased nearly 80%. This translated into a 50% decrease in hepatic CE concentration in conjunction with a rarely seen 2-fold increase in hepatic FC concentration. Acute hepatic ACAT2KD increased plasma FC levels by 25%, which subsided after two weeks of treatment. The increased plasma FC was primarily associated with large and small LDL. After one week of hepatic ACAT2KD mice had a minor, non-significant increase in biliary cholesterol levels but had a 2-fold increase in fecal neutral sterol (FNS) excretion. In summary our data show that when CE is rapidly cleared from the liver, FNS levels dramatically increase with a transient increase in hepatic and plasma FC levels. We believe that in the ACAT2KD mice the increased plasma FC associated with LDL is feeding into TICE thus resulting in increased FNS.