J. Lipid Res.
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Originally published In Press as doi:10.1194/jlr.M300162-JLR200 on July 1, 2003

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Journal of Lipid Research, Vol. 44, 1833-1840, October 2003
Copyright © 2003 by American Society for Biochemistry and Molecular Biology

Mobilization of cytoplasmic CE droplets by overexpression of human macrophage cholesteryl ester hydrolase

Shobha Ghosh1,*, Richard W. St. Clair{dagger} and Lawrence L. Rudel{dagger}

* Department of Internal Medicine, Virginia Commonwealth University, Richmond, VA 23298-0050
{dagger} Department of Pathology, Wake Forest University School of Medicine, Winston-Salem, NC 27157

1 To whom correspondence should be addressed. e-mail: shobha{at}hsc.vcu.edu

The obligatory first step in the removal of cholesterol from foam cells is the hydrolysis of stored cholesteryl esters (CEs) to release free cholesterol (FC). Neutral cholesteryl ester hydrolase (CEH) catalyzes this hydrolysis, and limiting levels of CEH could play a role in determining the susceptibility to atherosclerosis. We have recently reported the first identification and cloning of cDNA for human macrophage CEH. In the present study, we tested the hypothesis that systematically varied levels of overexpression of human macrophage CEH results in a proportional degree of reduction in cellular CE content in a cell system with known and reproducible amounts of CE accumulation. CEH expression was confirmed by demonstrating the presence of CEH mRNA and protein with an increase in CEH activity. A significant reduction in intracellular lipid droplets was observed in CEH-expressing cells, together with a decrease in cellular CE mass and a 2-fold increase in FC efflux.

These results demonstrate that when human macrophage CEH is expressed in lipid-laden cells, hydrolysis and mobilization of CE (stored as lipid droplets) occur. These data establish the possibility that increased CE hydrolysis, mediated by CEH up-regulation, could represent an important mechanism to reduce the cholesterol burden of foam cells.

Abbreviations: CE, cholesteryl ester; CEH, cholesteryl ester hydrolase; FC, free or unesterified cholesterol; PC, phosphatidylcholine

Supplementary key words reverse cholesterol transport • cholesterol efflux • atherosclerosis • macrophage foam cells • cholesteryl ester


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