Endogenous 24(S),25-epoxycholesterol fine-tunes acute control of cellular cholesterol homeostasis

Certain oxysterols, when added to cultured cells, are potent regulators of cholesterol homeostasis, decreasing cholesterol synthesis and uptake and increasing cholesterol efflux. However, very little is known about whether or not endogenous oxysterol(s) plays a significant role in cholesterol homeostasis. 24(S),25-Epoxycholesterol (24,25EC) is unique among oxysterols in that it is produced in a shunt of the mevalonate pathway which also produces cholesterol. We investigated the role of endogenously produced 24,25EC using a novel strategy of overexpressing the enzyme 2,3-oxidosqualene cyclase in Chinese hamster ovary cells to selectively inhibit the synthesis of this oxysterol. First, loss of 24,25EC decreased expression of the LXR target gene, ABCA1, substantiating its role as an endogenous ligand for LXR. Second, loss of 24,25EC increased acute cholesterol synthesis, which was rationalized by a concomitant increase in HMG-CoA reductase gene expression at the level of SREBP-2 processing. Therefore, in the absence of 24,25EC, fine-tuning of the acute regulation of cholesterol homeostasis is lost, supporting the hypothesis that 24,25EC functions to protect the cell against the accumulation of newly synthesized cholesterol.     

25-Hydroxycholesterol increases the availability of cholesterol in phospholipid membranes

Side-chain oxysterols are enzymatically generated oxidation products of cholesterol that serve a central role in mediating cholesterol homeostasis. Recent work has shown that side-chain oxysterols, such as 25-hydroxycholesterol (25-HC), alter membrane structure in very different ways from cholesterol, suggesting a possible mechanism for how these oxysterols regulate cholesterol homeostasis. Here we extend our previous work by using molecular-dynamics simulations of 25-HC and cholesterol mixtures in 1-palmitoyl-2-oleoyl-phosphatidylcholine bilayers to examine the combined effects of 25-HC and cholesterol in the same bilayer. 25-HC causes larger changes in membrane structure when added to cholesterol-containing membranes than when added to cholesterol-free membranes. We also find that the presence of 25-HC changes the position, orientation, and solvent accessibility of cholesterol, shifting it into the water interface and thus increasing its availability to external acceptors. This is consistent with experimental results showing that oxysterols can trigger cholesterol trafficking from the plasma membrane to the endoplasmic reticulum. These effects provide a potential mechanism for 25-HC-mediated regulation of cholesterol trafficking and homeostasis through modulation of cholesterol availability.     

Oxysterols, cholesterol homeostasis, and Alzheimer disease

Aberrant cholesterol metabolism has been implicated in Alzheimer disease (AD) and other neurological disorders. Oxysterols and other cholesterol oxidation products are effective ligands of liver X activated receptor (LXR) nuclear receptors, major regulators of genes subserving cholesterol homeostasis. LXR receptors act as molecular sensors of cellular cholesterol concentrations and effectors of tissue cholesterol reduction. Following their interaction with oxysterols, activation of LXRs induces the expression of ATP-binding cassette, sub-family A member 1, a pivotal modulator of cholesterol efflux. The relative solubility of oxysterols facilitates lipid flux among brain compartments and egress across the blood-brain barrier. Oxysterol-mediated LXR activation induces local apoE biosynthesis (predominantly in astrocytes) further enhancing cholesterol re-distribution and removal. Activated LXRs invoke additional neuroprotective mechanisms, including induction of genes governing bile acid synthesis (sterol elimination pathway), apolipoprotein elaboration, and amyloid precursor protein processing. The latter translates into attenuated beta-amyloid production that may ameliorate amyloidogenic neurotoxicity in AD brain. Stress-induced up-regulation of the heme-degrading enzyme, heme oxygenase-1 in AD-affected astroglia may impact central lipid homeostasis by promoting the oxidation of cholesterol to a host of oxysterol intermediates. Synthetic oxysterol-mimetic drugs that activate LXR receptors within the CNS may provide novel therapeutics for management of AD and other neurological afflictions characterized by deranged tissue cholesterol homeostasis.     

Oxysterols and redox signaling in the pathogenesis of non-alcoholic fatty liver disease

Abstract

Oxysterols are oxidized species of cholesterol coming from exogenous (e.g. dietary) and endogenous (in vivo) sources. They play critical roles in normal physiologic functions such as regulation of cellular cholesterol homeostasis. Most of biological effects are mediated by interaction with nuclear receptor LXRα, highly expressed in the liver as well as in many other tissues. Such interaction participates in the regulation of whole-body cholesterol metabolism, by acting as “lipid sensors”. Moreover, it seems that oxysterols are also suspected to play key roles in several pathologies, including cardiovascular and inflammatory disease, cancer, and neurodegeneration. Growing evidence suggests that oxysterols may contribute to liver injury in non-alcoholic fatty liver disease. The present review focuses on the current status of knowledge on oxysterols’ biological role, with an emphasis on LXR signaling and oxysterols’ physiopathological relevance in NAFLD, suggesting new pharmacological development that needs to be addressed in the near future.     

Oxysterols in human circulation: which role do they have?

Oxysterols are oxygenated derivatives of cholesterol that are intermediates in cholesterol excretion pathways. They may also be regarded as transport forms of cholesterol and introduction of an additional hydroxyl group facilitates flux of cholesterol across cell membranes and the blood-brain barrier. According to current concepts, oxysterols are also mediating a number of cholesterol-induced metabolic effects. The recent discovery of nuclear receptors with an affinity for oxysterols has given support to this concept. Nuclear receptors such as liver X receptor alpha do have a role in cholesterol homeostasis, but there is still only indirect evidence that oxysterols are the physiological ligands. In this overview we report some recent advancements in our knowledge about the origin and metabolic fate of the quantitatively most important oxysterols occurring in the circulation. In addition, we discuss the possibility that some of these oxysterols may activate liver X receptors and regulate cholesterol homeostasis.     

Cholesterol metabolism: from lipidomics to immunology

Oxysterols, the oxidized forms of cholesterol or of its precursors, are formed in the first steps of cholesterol metabolism. Oxysterols have interested chemists, biologists, and physicians for many decades, but their exact biological relevance in vivo, other than as intermediates in bile acid biosynthesis, has long been debated. However, in the first quarter of this century, a role for side-chain oxysterols and their C-7 oxidized metabolites has been convincingly established in the immune system. 25-Hydroxycholesterol has been shown to be synthesized by macrophages in response to the activation of Toll-like receptors and to offer protection against microbial pathogens, whereas 7α,25-dihydroxycholesterol has been shown to act as a chemoattractant to lymphocytes expressing the G protein-coupled receptor Epstein-Barr virus-induced gene 2 and to be important in coordinating the action of B cells, T cells, and dendritic cells in secondary lymphoid tissue. There is a growing body of evidence that not only these two oxysterols but also many of their isomers are of importance to the proper function of the immune system. Here, we review recent findings related to the roles of oxysterols in immunology.     

NPC1 and NPC2 regulate cellular cholesterol homeostasis through generation of low density lipoprotein cholesterol-derived oxysterols

Mutations in the Niemann-Pick disease genes cause lysosomal cholesterol accumulation and impaired low density lipoprotein (LDL) cholesterol esterification. These findings have been attributed to a block in cholesterol movement from lysosomes to the site of the sterol regulatory machinery. In this study we show that Niemann-Pick type C1 (NPC1) and Niemann-Pick type C2 (NPC2) mutants have increased cellular cholesterol, yet they are unable to suppress LDL receptor activity and cholesterol biosynthesis. Cholesterol overload in both NPC1 and NPC2 mutants results from the failure of LDL cholesterol tobothsuppresssterolregulatoryelement-bindingprotein-dependent gene expression and promote liver X receptor-mediated responses. However, the severity of the defect in regulation of sterol homeostasis does not correlate with endoplasmic reticulum cholesterol levels, but rather with the degree to which NPC mutant fibroblasts fail to appropriately generate 25-hydroxycholesterol and 27-hydroxycholesterol in response to LDL cholesterol. Moreover, we demonstrate that treatment with oxysterols reduces cholesterol in NPC mutants and is able to correct the NPC1I1061T phenotype, the most prevalent NPC1 disease genotype. Our findings support a role for NPC1 and NPC2 in the regulation of sterol homeostasis through generation of LDL cholesterol-derived oxysterols and have important implications for the treatment of NPC disease.     

Oxysterols: Sources,cellular storage and metabolism,and new insights into their roles in cholesterol homeostasis

Oxysterols are structurally identical to cholesterol, but with one or more additional oxygen containing functional groups (such as alcohol, carbonyl or epoxide groups). The wide array of oxysterols encountered in human health and disease vary in their origin (either enzymic or non-enzymic), and their putative effects and/or function(s). Some are thought to be damaging, whereas others may play important physiological roles, including in the regulation of cholesterol homeostasis. In this review, we will concentrate on the major cellular oxysterols. We summarise their location, generation, metabolism and elimination, as well as providing insights into the latest research into their regulatory roles in cholesterol homeostasis.     

Oxysterols: genesis and basic functions

The structural peculiarities of the most widespread oxysterols, the products of oxidative transformations of cholesterol are discussed. The transformations proceed with the participation of enzymatic systems of the body or as a result of various nonenzymatic reactions. The pathways of their formation from cholesterol are also considered. The role of oxysterols in the maintenance of cholesterol homeostasis and in the development of atherosclerosis is reviewed. The possibility of using oxysterols as markers of pathological processes is demonstrated.     

Oxysterols: Genesis and basic functions

The structural peculiarities of the most widespread oxysterols, the products of oxidative transformations of cholesterol are discussed. The transformations proceed with the participation of enzymatic systems of the body or as a result of various nonenzymatic reactions. The pathways of their formation from cholesterol are also considered. The role of oxysterols in the maintenance of cholesterol homeostasis and in the development of atherosclerosis is reviewed. The possibility of using oxysterols as markers of pathological processes is demonstrated.     

Biological activities of oxysterols

Literature dealing with the biological activities of cholesterol autoxidation products and related oxysterols in vivo and in vitro published since the previous 1981 monograph is reviewed. Although several oxysterols are important cholesterol metabolites implicated in bile acids and steroid hormones biosynthesis, effects on cellular membranes and on specific enzyme systems as well as cytotoxic, atherogenic, mutagenic, and carcinogenic activities characterize oxysterols as a class. Circumstantial evidence implicates oxysterols of the human diet and those formed in vivo with human health disorders, but recent work also supports an hypothesis that some oxysterols be endogenous intracellular regulators of de novo sterol biosynthesis. The true physiological relevance, if any, of these matters has not been adduced.     

Oxysterols: functional significance in fetal development and the maintenance of normal retinal function

PURPOSE OF REVIEW: Recent findings extend the biologic activities of oxysterols as ligands for nuclear receptors to a role in morphogenesis during fetal development and to a role in the metabolism of photooxidation products of cholesterol in the retina. RECENT FINDINGS: A 1000-fold increase of the 27-hydroxy metabolite of 7-dehydrocholesterol in the plasma of children with Smith-Lemli-Opitz syndrome imply that intermediates in cholesterol synthesis follow alternate pathways of metabolism that generate novel oxysterols. A mouse model also finds an increase in sterol intermediates as the proximate cause of dysmorphisms. A role for oxysterols in the effects of Sonic hedgehog protein focuses on their role in normal fetal development. Both CYP27A1 and CYP46A1 are expressed in primate retina indicating that local metabolism of 7-ketocholesterol to nontoxic derivatives is important for preventing retinal degeneration. SUMMARY: Recent data expand the functional roles of oxysterols to fetal development and to the detoxification of oxidation products of cholesterol. This review shifts the focus of attention from studies of their ligand-binding activity to studies of animal models that indicate a number of important biologic effects during fetal development and during the aging process.     

Oxysterols and mechanisms of apoptotic signaling: implications in the pathology of degenerative diseases

Oxysterols, or cholesterol oxidation products, are oxygenated derivatives of cholesterol which are formed endogenously during the biosynthesis of bile acids and steroid hormones. In addition, oxysterols may also be absorbed from the diet as they are found in many commonly consumed foods. Oxysterols have been shown to possess many potent and diverse biological activities, and the study of the effects of these oxidation products on the human body forms a wide field of research. The results of most research efforts support the conclusion that certain oxysterols, predominantly those found in oxidized low-density lipoprotein, exert pathological effects such as the induction of apoptotic cell death. Moreover, apoptosis induced by oxysterols has been strongly implicated in the pathogenesis of atherosclerosis as well as a variety of other diseases. The study of oxysterol-induced apoptosis is an emerging area, and the following review aims to provide a detailed account on the chronology of events involved. Current evidence of the involvement of the death receptor pathway and protein kinases is examined as well as important apoptosis regulators such as the mitochondria, B-cell lymphoma-2 proteins and caspases. The effect of oxysterols on gene expression, protein interactions and membrane properties are also discussed.     

Oxysterols: From cholesterol metabolites to key mediators

Oxysterols are cholesterol metabolites that can be produced through enzymatic or radical processes. They constitute a large family of lipids (i.e. the oxysterome) involved in a plethora of physiological processes. They can act through GPCR (e.g. EBI2, SMO, CXCR2), nuclear receptors (LXR, ROR, ERα) and through transporters or regulatory proteins. Their physiological effects encompass cholesterol, lipid and glucose homeostasis. Additionally, they were shown to be involved in other processes such as immune regulatory functions and brain homeostasis. First studied as precursors of bile acids, they quickly emerged as interesting lipid mediators. Their levels are greatly altered in several pathologies and some oxysterols (e.g. 4β-hydroxycholesterol or 7α-hydroxycholestenone) are used as biomarkers of specific pathologies. In this review, we discuss the complex metabolism and molecular targets (including binding properties) of these bioactive lipids in human and mice. We also discuss the genetic mouse models currently available to interrogate their effects in pathophysiological settings. We also summarize the levels of oxysterols reported in two key organs in oxysterol metabolism (liver and brain), plasma and cerebrospinal fluid. Finally, we consider future opportunities and directions in the oxysterol field in order to gain a better insight and understanding of the complex oxysterol system.