Role of STAR and SCP2/SCPx in the Transport of Cholesterol and Other Lipids

Cholesterol is a lipid molecule essential for several key cellular processes including steroidogenesis. As such, the trafficking and distribution of cholesterol is tightly regulated by various pathways that include vesicular and non-vesicular mechanisms. One non-vesicular mechanism is the binding of cholesterol to cholesterol transport proteins, which facilitate the movement of cholesterol between cellular membranes. Classic examples of cholesterol transport proteins are the steroidogenic acute regulatory protein (STAR; STARD1), which facilitates cholesterol transport for acute steroidogenesis in mitochondria, and sterol carrier protein 2/sterol carrier protein-x (SCP2/SCPx), which are non-specific lipid transfer proteins involved in the transport and metabolism of many lipids including cholesterol between several cellular compartments. This review discusses the roles of STAR and SCP2/SCPx in cholesterol transport as model cholesterol transport proteins, as well as more recent findings that support the role of these proteins in the transport and/or metabolism of other lipids.     

Should the forms of dietary choline also be considered when estimating dietary intake and the implications for health?

Although epidemiological studies suggest that populations are not meeting daily recommendations for total choline, they fail to consider the forms of choline in the diet. Aqueous and lipid‐soluble choline forms differ in absorption, metabolism and functions, suggesting that the form matters. It has been demonstrated that the relative amount of these forms of choline can be altered through consuming different foods and using different preparation methods. Evidence suggests that the forms of choline may differentially impact growth, immune function, plasma and serum cholesterol levels and brain development, therefore in addition to estimating the amount total choline in the diet, researchers should also consider what forms of choline are consumed in the diet.     

NGF Modulates Cholesterol Metabolism and Stimulates ApoE Secretion in Glial Cells Conferring Neuroprotection against Oxidative Stress

Cholesterol plays a crucial role in the brain, where its metabolism is particularly regulated by astrocytic activity. Indeed, adult neurons suppress their own cholesterol biosynthesis and import this sterol through ApoE-rich particles secreted from astrocytes. Recent evidence suggests that nerve growth factor (NGF) may exert neurotrophic activity by influencing cell metabolism. Nevertheless, the effect of NGF on glial cholesterol homeostasis has still not been elucidated. Thus, the aim of this project is to assess whether NGF could influence cholesterol metabolism in glial cells. To reach this objective, the U373 astrocyte-derived cell line was used as an experimental model. Immunoblot and ELISA analysis showed that proteins and enzymes belonging to the cholesterol metabolism network were increased upon NGF treatment in glial cells. Furthermore, NGF significantly increased ApoE secretion and the amount of extracellular cholesterol in the culture medium. Co-culture and U373-conditioned medium experiments demonstrated that NGF treatment efficiently counteracted rotenone-mediated cytotoxicity in N1E-115 neuronal cells. Conversely, neuroprotection mediated by NGF treatment was suppressed when N1E-115 were co-cultured with ApoE-silenced U373 cells. Taken together, these data suggest that NGF controls cholesterol homeostasis in glial cells. More importantly, NGF exerts neuroprotection against oxidative stress, which is likely associated with the induction of glial ApoE secretion.     

Myelin and multiple sclerosis

Myelin is a lipid rich membrane system elaborated by oligodendrocytes in the central nervous system and Schwann cells in the peripheral nervous system to provide axons with a high resistance but discontinuous sheath. The node internode structure that results allows very fast saltatory nerve impulse conduction. Myelin is rich in cholesterol, phospholipids and glycolipids. However, the ability of the brain to synthesise cholesterol determines its capacity to elaborate myelin and as a result cholesterol is an essential nutrient for brain function. Multiple sclerosis is an autoimmune disease of the brain and nervous system resulting in inflammation leading to demyelination and eventually to axonal degeneration. The discovery of remyelination a regenerative process offers some hope that by boosting its capacity more effective treatments for multiple sclerosis will be developed in the future.     

Brain Energy Metabolism in Ischemic Stroke: Effects of Smoking and Diabetes

Proper regulation of energy metabolism in the brain is crucial for maintaining brain activity in physiological and different pathophysiological conditions. Ischemic stroke has a complex pathophysiology which includes perturbations in the brain energy metabolism processes which can contribute to worsening of brain injury and stroke outcome. Smoking and diabetes are common risk factors and comorbid conditions for ischemic stroke which have also been associated with disruptions in brain energy metabolism. Simultaneous presence of these conditions may further alter energy metabolism in the brain leading to a poor clinical prognosis after an ischemic stroke event. In this review, we discuss the possible effects of smoking and/or diabetes on brain glucose utilization and mitochondrial energy metabolism which, when present concurrently, may exacerbate energy metabolism in the ischemic brain. More research is needed to investigate brain glucose utilization and mitochondrial oxidative metabolism in ischemic stroke in the presence of smoking and/or diabetes, which would provide further insights on the pathophysiology of these comorbid conditions and facilitate the development of therapeutic interventions.     

Fast Tacrolimus Metabolism Does Not Promote Post-Transplant Diabetes Mellitus after Kidney Transplantation

Post-transplant diabetes mellitus (PTDM) after kidney transplantation induced by tacrolimus is an important issue. Fast tacrolimus metabolism, which can be estimated by concentration-to-dose (C/D) ratio, is associated with increased nephrotoxicity and unfavorable outcomes after kidney transplantation. Herein, we elucidate whether fast tacrolimus metabolism also increases the risk for PTDM. Data from 596 non-diabetic patients treated with tacrolimus-based immunosuppression at the time of kidney transplantation between 2007 and 2015 were retrospectively analyzed. The median follow-up time after kidney transplantation was 4.7 years (IQR 4.2 years). Our analysis was complemented by experimental modeling of fast and slow tacrolimus metabolism kinetics in cultured insulin-producing pancreatic cells (INS-1 cells). During the follow-up period, 117 (19.6%) patients developed PTDM. Of all patients, 210 (35.2%) were classified as fast metabolizers (C/D ratio < 1.05 ng/mL × 1/mg). Fast tacrolimus metabolizers did not have a higher incidence of PTDM than slow tacrolimus metabolizers (p = 0.496). Consistent with this, insulin secretion and the viability of tacrolimus-treated INS-1 cells exposed to 12 h of tacrolimus concentrations analogous to the serum profiles of fast or slow tacrolimus metabolizers or to continuous exposure did not differ (p = 0.286). In conclusion, fast tacrolimus metabolism is not associated with increased incidence of PTDM after kidney transplantation, either in vitro or in vivo. A short period of incubation of INS-1 cells with tacrolimus using different concentration profiles led to comparable effects on cell viability and insulin secretion in vitro. Consistent with this, in our patient, collective fast Tac metabolizers did not show a higher PTDM incidence compared to slow metabolizers.     

Relationship between Brain Metabolic Disorders and Cognitive Impairment: LDL Receptor Defect

The low-density-lipoprotein receptor (LDLr) removes low-density lipoprotein (LDL), an endovascular transporter that carries cholesterol from the bloodstream to peripheral tissues. The maintenance of cholesterol content in the brain, which is important to protect brain function, is affected by LDLr. LDLr co-localizes with the insulin receptor and complements the internalization of LDL. In LDLr deficiency, LDL blood levels and insulin resistance increase, leading to abnormal cholesterol control and cognitive deficits in atherosclerosis. Defects in brain cholesterol metabolism lead to neuroinflammation and blood–brain-barrier (BBB) degradation. Moreover, interactions between endoplasmic reticulum stress (ER stress) and mitochondria are induced by ox-LDL accumulation, apolipoprotein E (ApoE) regulates the levels of amyloid beta (Aβ) in the brain, and hypoxia is induced by apoptosis induced by the LDLr defect. This review summarizes the association between neurodegenerative brain disease and typical cognitive deficits.     

Hepatic Expression of Long-Chain Acyl-CoA Synthetase 3 is Upregulated in Hyperlipidemic Hamsters

Members of the mammalian long-chain acyl-CoA synthetase (ACSL) family are key enzymes for cellular fatty acid metabolism that catalyze the initial step in activation of long-chain fatty acids. However, the specificity of individual isoforms of ACSL to the lipid metabolic process is not well studied. In addition, the regulation of expression of individual ACSL isoforms under hyperlipidemic conditions is largely unknown. We cloned the hamster ACSL3 cDNA coding region and generated specific antibodies recognizing the ACSL3 protein. We next observed the changes in ACSL3 mRNA and protein expression in hamsters fed a standard chow diet or a high fat and high cholesterol (HFHC) diet. HFHC feeding significantly increased ACSL3 mRNA and protein expression in liver and to a lesser extent in muscle but not in adipose, brain, heart, or testis. Additionally, ACSL3 mRNA abundance was differentially regulated by the nutritional status in different tissues with liver, muscle, and adipose being the most sensitive tissues. Importantly, the hepatic ACSL3 mRNA expression pattern in response to fasting and refeeding in hyperlipidemic hamsters differed from that observed in normal chow-fed hamsters. Together, these results provide the first in vivo evidence of altered regulation of hepatic ACSL3 expression under hyperlipidemic conditions and suggest important regulatory roles for this enzyme in lipid metabolism.     

Different thermal behavior of neonatal hepatic and cerebral 3-hydroxy-3-methylglutaryl-CoA reductase

The Arrhenius plots of hepatic and cerebral 3-hydroxy-3-methylglutaryl-CoA reductase activity were studied in neonatal chicks fed with a standard diet. Supplementation of the diet with 2% cholesterol from hatching has no effect on the thermal characteristics of the brain enzyme. The Arrhenius plot of brain reductase was practically similar to that found in control chicks. However, hepatic reductase was inhibited by cholesterol feeding. Dietary cholesterol increased the cholesterol/lipidic phosphorus molar ratio in liver microsomes, whereas no significant differences were observed in brain microsomes. These results are in agreement with the hypothesis that activity of hepatic reductase is regulated by the fluidity of microsomal membrane and show that cholesterol feeding does not alter the fluidity of microsomal membranes in neonatal chick brain having, thus, no effect on the thermal behavior of cerebral reductase.     

Activation of acyl-CoA cholesterol acyltransferase: Redistribution in microsomal fragments of cholesterol and its facilitated movement by methyl-β-cyclodextrin

Acyl-CoA cholesterol acyltransferase (ACAT) (EC 2,3,1.26) in the yolk sac membrane of chicken eggs plays an important role in the transport of lipids, which serve as both structural components and as an energy source during embryogenesis. ACAT from the yolk sac membrane of chicken eggs 16 d after fertilization has higher activity and better stability than its mammalian liver counterpart. During our study of the avian enzyme, ACAI was found to be activated up to twofold during storage at 4°C. The activation was investigated, and data suggest that redistribution of cholesterol within microsomal vesicles leads to the increase. Methyl-β-cyclodextrin (MβCD) increases activation an additional twofold, possibly by facilitating the movement of cholesterol within microsomal fragments and allowing redistribution of cholesterol in lipid bilayers to a greater extent. Treatment of microsomes with MβCD removes cholesterol from the membranes. Controlled amounts of cholesterol can be restored to the membranes by mixing them with cholesterol-phosphatidylcholine liposomes in the presence of MβCD. Under these conditions, the plot of ACAT vs. cholesterol mole fraction in the liposomes is sigmoidal. The finding that MβCD can enhance cholesterol transfer between liposomes and microsomes and reduce the limitation of slow movement et nonpolar molecules in aqueous media should make cyclodextrins more useful in in vitro studies of apolar molecule transport between membrane vesicles.     

Reduced Levels of ABCA1 Transporter Are Responsible for the Cholesterol Efflux Impairment in β-Amyloid-Induced Reactive Astrocytes: Potential Rescue from Biomimetic HDLs

The cerebral synthesis of cholesterol is mainly handled by astrocytes, which are also responsible for apoproteins’ synthesis and lipoproteins’ assembly required for the cholesterol transport in the brain parenchyma. In Alzheimer disease (AD), these processes are impaired, likely because of the astrogliosis, a process characterized by morphological and functional changes in astrocytes. Several ATP-binding cassette transporters expressed by brain cells are involved in the formation of nascent discoidal lipoproteins, but the effect of beta-amyloid (Aβ) assemblies on this process is not fully understood. In this study, we investigated how of Aβ_1-42-induced astrogliosis affects the metabolism of cholesterol in vitro. We detected an impairment in the cholesterol efflux of reactive astrocytes attributable to reduced levels of ABCA1 transporters that could explain the decreased lipoproteins’ levels detected in AD patients. To approach this issue, we designed biomimetic HDLs and evaluated their performance as cholesterol acceptors. The results demonstrated the ability of apoA-I nanodiscs to cross the blood–brain barrier in vitro and to promote the cholesterol efflux from astrocytes, making them suitable as a potential supportive treatment for AD to compensate the depletion of cerebral HDLs.     

Aging Induces Tissue-Specific Changes in Cholesterol Metabolism in Rat Brain and Liver

Disturbance of cholesterol homeostasis in the brain is coupled to age-related brain dysfunction. In the present work, we studied the relationship between aging and cholesterol metabolism in two brain regions, the cortex and hippocampus, as well as in the sera and liver of 6-, 12-, 18- and 24-month-old male Wistar rats. Using gas chromatography-mass spectrometry, we undertook a comparative analysis of the concentrations of cholesterol, its precursors and metabolites, as well as dietary-derived phytosterols. During aging, the concentrations of the three cholesterol precursors examined (lanosterol, lathosterol and desmosterol) were unchanged in the cortex, except for desmosterol which decreased (44 %) in 18-month-old rats. In the hippocampus, aging was associated with a significant reduction in lanosterol and lathosterol concentrations at 24 months (28 and 25 %, respectively), as well as by a significant decrease of desmosterol concentration at 18 and 24 months (36 and 51 %, respectively). In contrast, in the liver we detected age-induced increases in lanosterol and lathosterol concentrations, and no change in desmosterol concentration. The amounts of these sterols were lower than in the brain regions. In the cortex and hippocampus, desmosterol was the predominant cholesterol precursor. In the liver, lathosterol was the most abundant precursor. This ratio remained stable during aging. The most striking effect of aging observed in our study was a significant decrease in desmosterol concentration in the hippocampus which could reflect age-related reduced synaptic plasticity, thus representing one of the detrimental effects of advanced age.     

Spontaneous crowding of ribosomes and proteins inside vesicles: a possible mechanism for the origin of cell metabolism

One of the open questions in the origin of life is the spontaneous formation of primitive cell-like compartments from free molecules in solution and membranes. "Metabolism-first" and "replicator-first" theories claim that early catalytic cycles first evolved in solution, and became encapsulated inside lipid vesicles later on. "Compartment-first" theories suggest that metabolism progressively occurred inside compartments. Both views have some weaknesses: the low probability of co-entrapment of several compounds inside the same compartment, and the need to control nutrient uptake and waste release, respectively. By using lipid vesicles as early-cell models, we show that ribosomes, proteins and lipids spontaneously self-organise into cell-like compartments to achieve high internal concentrations, even when starting from dilute solutions. These findings suggest that the assembly of cell-like compartments, despite its low probability of occurrence, is indeed a physically realistic process. The spontaneous achievement of high local concentration might provide a rational account for the origin of primitive cellular metabolism.     

Myelin Defects in Niemann–Pick Type C Disease: Mechanisms and Possible Therapeutic Perspectives

Niemann–Pick type C (NPC) disease is a wide-spectrum clinical condition classified as a neurovisceral disorder affecting mainly the liver and the brain. It is caused by mutations in one of two genes, NPC1 and NPC2, coding for proteins located in the lysosomes. NPC proteins are deputed to transport cholesterol within lysosomes or between late endosome/lysosome systems and other cellular compartments, such as the endoplasmic reticulum and plasma membrane. The first trait of NPC is the accumulation of unesterified cholesterol and other lipids, like sphingosine and glycosphingolipids, in the late endosomal and lysosomal compartments, which causes the blockade of autophagic flux and the impairment of mitochondrial functions. In the brain, the main consequences of NPC are cerebellar neurodegeneration, neuroinflammation, and myelin defects. This review will focus on myelin defects and the pivotal importance of cholesterol for myelination and will offer an overview of the molecular targets and the pharmacological strategies so far proposed, or an object of clinical trials for NPC. Finally, it will summarize recent data on a new and promising pharmacological perspective involving A_2A adenosine receptor stimulation in genetic and pharmacological NPC dysmyelination models.     

Oil-binding capacity of plastic fats: Effects of intermediate melting point TAG

The oil-binding capacity (OBC) of fat crystals was investigated as a function of intermediate melting point TAG (IMP-TAG) and stearin composition. Samples were prepared by melting 14% hard fraction (palm-canola stearin and IMP-TAG blends) in 86% liquid oil (olive, canola, safflower, and triolein) and crystallizing the mixture under fast and slow cooling conditions. Under fast cooling conditions, the OBC increased as the IMP-TAG/stearin ratio increased. The OBC is the grams of bound oil (determined by centrifugation) divided by the grams of solid fat (determined by pulse NMR). The maximum OBC was observed at 14% IMP-TAG and 0% stearin. In constrast, under slow cooling conditions, the 14% IMP-TAG and 0% stearin sample did not form crystals, and only free oil was present. The OBC for each liquid oil was lower under slow cooling conditions than under fast cooling conditions when compared at the same IMP-TAG/ stearin ratio. Solid fat content and RP-HPLC analyses indicated that IMP-TAG were retained in the crystal structure when processed under fast cooling conditions. RP-HPLC analyses also revealed that TAG with two saturated FA were retained in the crystal structure and that the monosaturated TAG were not. It was concluded that the TAG composition and cooling conditions played an important role in determining the OBC.     

Lipids and the pill

The widespread use of anovulatory compounds and the well-known effects of sex hormones on various aspects of metabolism prompted this review of our work and the work of others on observed changes in lipid metabolism resulting from the administration of oral contraceptives and their components. In the rat, female sex hormone administration results in a decreased plasma cholesterol level, an accumulation of cholesterol in liver and a decreased hepatic cholesterol biosynthesis. On the other hand, cholesterol biosynthesis is enhanced in ovaries and adrenals. There is also a diminished alpha lipoprotein content and a corresponding decrease in the alpha/beta lipoprotein ratio. In some cases these changes are comparable to those observed during pregnancy. The results of sex hormone administration to women are more variable. In this case the most often observed effect is hypertriglyceridemia. Changes in lipoprotein content and distribution are also evident and may be the result of changes in metabolism in the liver, e.g., lipid synthesis or lipid transport from liver to plasma and tissues, or both. Many of these changes may be mediated indirectly through the action of estrogenprogestin on other hormones. In both species the effects of oral contraceptives are attributable principally to the estrogen component. The combination of estrogen with progestin compounds, which constitutes the oral contraceptive, modifies the effects of estrogen administered alone.