Lipid-poor Apolipoprotein Research Articles

Conclusions from the VA-HIT study

Conclusions from the VA-HIT study

ABCA1-mediated transport of cellular cholesterol and phospholipids to HDL apolipoproteins.

Lipid-poor apolipoproteins remove cellular cholesterol and phospholipids by an active transport pathway controlled by an ATP binding cassette transporter called ABCA1 (formerly ABC1). Mutations in ABCA1 cause Tangier disease, a severe HDL deficiency syndrome characterized by a rapid turnover of plasma apolipoprotein A-I, accumulation of sterol in tissue macrophages, and prevalent atherosclerosis. This implies that lipidation of apolipoprotein A-I by the ABCA1 pathway is required for generating HDL particles and clearing sterol from macrophages. Thus, the ABCA1 pathway has become an important therapeutic target for mobilizing excess cholesterol from tissue macrophages and protecting against atherosclerosis.

Cubilin, a high-density lipoprotein receptor.

The metabolism of HDL particles is a complex biological process involving various regulating factors in plasma and different cellular receptors. In addition to the well-established scavenger receptor BI-mediated selective HDL-cholesteryl ester uptake in liver and steroidogenic tissues, evidence has been provided that HDL also undergoes holoparticle endocytosis in different tissues. Recently, a novel receptor expressed in various absorptive epithelia was disclosed as a high affinity receptor for endocytosis of HDL and lipid-poor apolipoprotein AI. This receptor, designated cubilin, may play an important role in the renal clearance of filterable apolipoprotein AI/HDL and in the maternal-fetal transport of cholesterol.

Apolipoprotein-mediated removal of cellular cholesterol and phospholipids

It is widely believed that high density lipoprotein (HDL) protects against cardiovascular disease by removing excess cholesterol from cells of the artery wall. Recent cell culture studies have provided evidence that a major pathway for removing cholesterol and phospholipids from cells is mediated by the direct interactions of HDL apolipoproteins (apo) with plasma membrane domains. These interactions efficiently clear cells of excess sterol by targeting for removal pools of cholesterol that feed into the cholesteryl ester cycle. The precursors for this pathway in vivo are likely to be lipid-free or lipid-poor apolipoproteins generated either by dissociation from the surface of HDL particles or by de novo synthesis. Fibroblasts from subjects with a severe HDL deficiency syndrome called Tangier disease have a cellular defect that prevents apolipoproteins from removing both cholesterol and phospholipids from cells. This defect is associated with a near absence of plasma HDL, markedly below normal low density lipoprotein (LDL) levels, and the appearance of macrophage foam cells in tissues. Thus, an inability of nascent apoA-I to acquire cellular lipids results in a rapid clearance of apoA-I from the plasma, decreased production and increased clearance of LDL, and sterol deposition in tissue macrophages. Although the molecular properties of this pathway are still poorly understood, these studies imply that the apolipoprotein-mediated pathway for removal of cellular lipids is a major source of plasma cholesterol and phospholipids and plays an important role in clearing excess cholesterol from macrophages in vivo.

Apolipoprotein E Produced by Human Monocyte-derived Macrophages Mediates Cholesterol Efflux That Occurs in the Absence of Added Cholesterol Acceptors

Human monocyte-derived macrophages can efflux accumulated cholesterol without exogenously added cholesterol acceptors (Kruth, H. S., Skarlatos, S. I., Gaynor, P. M., and Gamble, W. (1994) J. Biol. Chem. 269, 24511-24518). Most of the effluxed cholesterol accumulates in the medium as apolipoprotein E-discoidal lipid particles. In the current study, we determined whether and to what degree cholesterol efflux from human monocyte-macrophages depended on apolipoprotein E secretion. Unexpectedly, 2-week-old differentiated monocyte-macrophages secreted similar amounts of apolipoprotein E without or with cholesterol enrichment. Apolipoprotein E mRNA levels in these macrophages were not increased by cholesterol enrichment and were comparable with levels in HepG2 cells. Without cholesterol enrichment, monocyte-macrophages secreted lipid-poor apolipoprotein E with a density >1.21 g/ml. By contrast, cholesterol enrichment of monocyte-macrophages induced the association of apoE with phospholipid and cholesterol to form discoidal particles that floated at densities of 1.08-1.10 g/ml. An anti-apolipoprotein E monoclonal antibody added to the culture medium significantly inhibited cholesterol and phospholipid efflux from the monocyte-macrophages. This showed that apolipoprotein E was required for most of the cholesterol efflux, and that apolipoprotein E did not leave macrophages with lipid but rather associated with lipid after it was secreted. Thus, 1) apolipoprotein E was constitutively secreted by differentiated human monocyte-macrophages, 2) apolipoprotein E only formed discoidal particles following macrophage cholesterol enrichment, 3) apolipoprotein E was necessary for cholesterol efflux to occur in the absence of added cholesterol acceptors and, in addition 4) the level of macrophage unesterified cholesterol was not rate-limiting for this cholesterol efflux, and 5) net phospholipid synthesis occurred in macrophages secondary to apoE-mediated loss of macrophage phospholipid. In conclusion, apolipoprotein E functions in an autocrine pathway that mediates cholesterol efflux from human monocyte-derived macrophages.

Lipid-poor apolipoprotein A-I in Hep G2 cells: formation of lipid-rich particles by incubation with dimyristoylphosphatidylcholine

Apolipoprotein A-I is a major secretory product of the human hepatoma cell line, Hep G2; approx. 70% of apolipoprotein A-I was separated from the medium as lipid-poor apolipoprotein A-I in the d > 1.21 g/ ml fraction while 30% was associated with high-density lipoproteins (HDL) of d 1.063–1.21 g/ ml. The lipid-poor apolipoprotein A-I contains 50% proapolipoprotein A-I which is similar to the isoform distribution in Hep G2 preformed HDL. We tested the ability of lipid-poor apolipoprotein A-I from Hep G2 to form complexes with dimyristoylphosphatidylcholine (DMPC) vesicles at DMPC/apolipoprotein A-I molar ratios of 100:1 and 300:1. Lipid-poor apolipoprotein A-I formed complexes which floated at d < 1.21 g/ ml; at a 100:1 ratio, 42.7 ± 9.4% of the apolipoprotein A-I was recovered in complex form while at a 300:1 ratio, 68.8 ± 6.3% was recovered. On electron microscopy, the former complexes were small discs 16.9 nm ± 4.5 S.D. in diameter while the latter were larger discs 21.4 ± 4.4 nm diameter. Non-denaturing gradient gel electrophoresis of complexes formed at a 100:1 ratio had a peak in the region corresponding to 9.64 ± 0.08 nm; these particles possessed two apolipoprotein A-I molecules. At the higher ratio, 300:1, two distinct complexes were identifiable, one which banded in the 9.7 nm region and the other in the 16.9–18.7 nm region. The former particles contained two molecules of apolipoprotein A-I and the latter, three molecules. This study demonstrates that lipid-poor apolipoprotein A-I which is rich in more basic isoforms forms discrete lipoprotein complexes similar to those formed by mature apolipoprotein A-I. It is further suggested that, under the appropriate conditions, precursor or nascent HDL may be assembled extracellularly.

Nutrition and biogenesis of plasma lipoproteins in nonhuman primates

In this article we examine the production of very low-density lipoprotein (VLDL) by perfused livers obtained from chow- and cholesterol-fed nonhuman primates. These data illustrate two important features of VLDL production. First, VLDL is secreted from the liver in a form very close to that of its plasma counterpart. Thus for chow-fed animals, plasma VLDL and liver perfusate VLDL have similar lipid compositions. Second, the composition of VLDL can be modified significantly by diet in each of two primate species, the Rhesus monkey and the baboon. Rhesus monkey livers uniformly secrete larger quantities of VLDL and show more dramatic dietary effects than do baboon livers. Nevertheless, perfused livers from both species reveal qualitatively similar responses to dietary peanut oil and to lard fed in combination with cholesterol. Both fat-containing diets induce the livers to secrete VLDL enriched in cholesteryl ester compared with control perfusates yet still cholesteryl ester deficient compared with the animals' plasma VLDL. Peanut oil diet reduces the hepatic output of VLDL-associated apoprotein B and triglyceride, whereas lard increases hepatic secretion of VLDL-associated lipids and apoprotein E. We conclude that the nature of dietary fat plays an important role in determining the profile and composition of lipoproteins formed and secreted by the primate liver. We have also briefly reviewed the production of high-density lipoprotein, which is probably formed in the plasma from many sources, with special emphasis on the possible role of newly secreted lipid-poor apolipoproteins A-I and A-II.