Bile Acid Precursors Research Articles

Innovative Target Therapies Are Able to Block the Inflammation Associated with Dysfunction of the Cholesterol Biosynthesis Pathway.

The cholesterol pathway is an essential biochemical process aimed at the synthesis of bioactive molecules involved in multiple crucial cellular functions. The end products of this pathway are sterols, such as cholesterol, which are essential components of cell membranes, precursors of steroid hormones, bile acids and other molecules such as ubiquinone. Several diseases are caused by defects in this metabolic pathway: the most severe forms of which cause neurological involvement (psychomotor retardation and cerebellar ataxia) as a result of a variety of cellular impairments, including mitochondrial dysfunction. These pathologies are induced by convergent mechanisms in which the mitochondrial unit plays a pivotal role contributing to defective apoptosis, autophagy and mitophagy processes. Unraveling these mechanisms would contribute to the development of effective drug treatments for these disorders. In addition, the development of biochemical models could have a substantial impact on the understanding of the mechanism of action of drugs that act on this pathway in multifactor disorders. In this review we will focus in particular on inhibitors of cholesterol synthesis, mitochondria-targeted drugs and inhibitors of the inflammasome.

Deficiency of calcium-independent phospholipase A2 beta with aging causes biliary epithelial ductular reaction associated with increased bile acids in enterohepatic circulation

Background/Aims: Calcium-independent phospholipase A2 beta (iPLA2beta) generates lysophosphatidylcholine which is recognized as a macrophage 'fine me' signal for removal of apoptotic cells. We have shown that iPLA2beta deficiency causes an accumulation of apoptotic hepatocytes associated with a decrease of serum and hepatic lipids including cholesterol. We here determined whether iPLA2beta deficiency in aged mice can result in abnormalities in the hepatobiliary system and the alterations of bile acid contents. Methods: Control (WT) and whole body male iPLA2beta-/- mice at 19 – 22 months old were used. Bile acid profiles were measured by liquid-chromatography mass spectrometry. Functional assays included qRT-PCR, histology, and immunohistochemistry (IHC). Results: Livers of aged iPLA2beta-/- mice exhibited increased IHC staining of α-smooth muscle actin and biliary epithelial marker cytokeratin 19 compared with those of aged WT mice. This was concomitant with increases of total, hydrophilic, hydrophobic and secondary bile acids in the bile. The increases of hydrophobic secondary bile acids such as deoxycholic and chenodeoxycholic acids were observed in liver and intestine of iPLA2beta-/- mice, which was associated with increased mRNA expression of CYP7A1 and CYP8B1, and decreased expression of bile acid export genes BSEP and MRP3 in liver as well as OSTα and OSTβ in intestine. In the feces of mutant mice, we observed an increase of deoxycholic acid (suggesting increased dehydroxylation by luminal bacteria), but decreases of tauro-ursodeoxycholic acid as well as tauro- and glyco-chenodeoxycholic acids (suggesting increased deconjugation by luminal bacteria). Conclusions: iPLA2beta deficiency in aged mice caused biliary ductular expansion, and an accumulation of bile acids within enterohepatic circulation. This accumulation was a result of suppressed hepatic and intestinal bile acid export and increased bile acid syntheses, and the latter may lead to a decreased level of the bile acid precursor cholesterol. Thus, iPLA2beta plays a homeostatic role by regulating hepatic bile acid metabolism affecting systemic cholesterol levels and intestinal bile acid contents.

59 LIPID DYNAMICS IN THE EARLY EMBRYONIC DEVELOPMENT OF ZEBRAFISH BY DESORPTION ELECTROSPRAY IONIZATION MASS SPECTROMETRY IMAGING AND NANOELECTROSPRAY–MASS SPECTROMETRY

This study introduces a combination of mass spectrometry-based analytical approaches with little or no sample preparation for the study of zebrafish (Danio rerio) lipid metabolism during early development (0 through 96 h post-fertilization, hpf). Desorption electrospray ionization mass spectrometry (DESI-MS) imaging and nanoelectrospray (nESI) MS were used. Embryos (N = 107) were placed onto filter paper squares and analysed by DESI-MS imaging. Embryos were arranged in rows with each one corresponding to a different developmental time, so that each DESI-MS image contained arrays of samples at different hpf. After completion of DESI-MS acquisition, the arrays were cut into strips of paper, isolating each embryo, which was subsequently inserted into a nESI tip and analysed. All experiments were performed using a linear ion trap mass spectrometer. DESI-MS images were acquired in the positive ion mode using acetonitrile doped with 10 ppm AgNO3, which allowed for the detection of cytosolic neutral lipids, such as cholesteryl esters (CE), diacylglicerols (DAG), triacylglycerols (TAG), squalene, and ubiquinone. For the nESI, experiments were run in full mass scan and MS/MS, in both negative and positive ion mode, using a mixture of dimethylformamide and acetonitrile to detect membrane phospholipids (e.g. phosphatidylcholines and phosphatidylglycerols). Principal component analysis was used to explore DESI-MS images and nESI data in an unsupervised fashion. The combination of DESI-MS imaging, providing chemical and spatial location, and nanoESI, providing a broader and more extensive structural information, showed that the lipid content changes dramatically over the first days of development. Accumulation of DAG and TAG, which are usually concentrated in the embryos yolk sac as an important storage for early development, were observed between 0 and 48 hpf. Embryos at the intermediate phase of development (24 hpf) were distributed between those of 0 and 48 hpf, reflecting the dynamics associated with development and might be correlated with TAG consumption and de novo synthesis. After 72 and 96 hpf, samples differed from prior developmental stages by the content of squalene, ubiquinone, and CE 22 : 5 (overexpressed), while the TAG content decreased. Further decrease of TAGs and ubiquinone occurred between 72 and 96 hpf. These observations indicated that zebrafish embryos rely entirely on the yolk sac for the nutrients needed to sustain growth and survival during the first four days of development. Yolk lipids are the likely source of TAGs, as well as cholesterol, a required component of cell membranes and a precursor for bile acids. The abundance of ubiquinone can be tied to zebrafish embryo growth, differentiation and organogenesis, and activation of mitochondria.

In-Depth Dissection of the P133R Mutation in Steroid 5β-Reductase (AKR1D1): A Molecular Basis of Bile Acid Deficiency.

Human steroid-5β-reductase (aldo-keto reductase 1D1, AKR1D1) stereospecifically reduces Δ(4)-3-ketosteroids to 5β-dihydrosteroids and is essential for steroid hormone metabolism and bile acid biosynthesis. Genetic defects in AKR1D1 cause bile acid deficiency that leads to life threatening neonatal hepatitis and cholestasis. The disease-associated P133R mutation caused significant decreases in catalytic efficiency with both the representative steroid (cortisone) and the bile acid precursor (7α-hydroxycholest-4-en-3-one) substrates. Pro133 is a second shell residue to the steroid binding channel and is distal to both the cofactor binding site and the catalytic center. Strikingly, the P133R mutation caused over a 40-fold increase in Kd values for the NADP(H) cofactors and increased the rate of release of NADP(+) from the enzyme by 2 orders of magnitude when compared to the wild type enzyme. By contrast the effect of the mutation on Kd values for steroids were 10-fold or less. The reduced affinity for the cofactor suggests that the mutant exists largely in the less stable cofactor-free form in the cell. Using stopped-flow spectroscopy, a significant reduction in the rate of the chemical step was observed in multiple turnover reactions catalyzed by the P133R mutant, possibly due to the altered position of NADPH. Thus, impaired NADPH binding and hydride transfer is the molecular basis for bile acid deficiency in patients with the P133R mutation. Results revealed that optimal cofactor binding is vulnerable to distant structural perturbation, which may apply to other disease-associated mutations in AKR1D1, all of which occur at conserved residues and are unstable.

Simultaneous determination of oxysterols, cholesterol and 25-hydroxy-vitamin D3 in human plasma by LC-UV-MS.

BackgroundOxysterols are promising biomarkers of neurodegenerative diseases that are linked with cholesterol and vitamin D metabolism. There is an unmet need for methods capable of sensitive, and simultaneous quantitation of multiple oxysterols, vitamin D and cholesterol pathway biomarkers.MethodsA method for simultaneous determination of 5 major oxysterols, 25-hydroxy vitamin D3 and cholesterol in human plasma was developed. Total oxysterols were prepared by room temperature saponification followed by solid phase extraction from plasma spiked with deuterated internal standards. Oxysterols were resolved by reverse phase HPLC using a methanol/water/0.1% formic acid gradient. Oxysterols and 25-hydroxy vitamin D3 were detected with atmospheric pressure chemical ionization mass spectrometry in positive ion mode; in-series photodiode array detection at 204nm was used for cholesterol. Method validation studies were performed. Oxysterol levels in 220 plasma samples from healthy control subjects, multiple sclerosis and other neurological disorders patients were quantitated.ResultsOur method quantitated 5 oxysterols, cholesterol and 25-hydroxy vitamin D3 from 200 μL plasma in 35 minutes. Recoveries were >85% for all analytes and internal standards. The limits of detection were 3-10 ng/mL for oxysterols and 25-hydroxy vitamin D3 and 1 μg/mL for simultaneous detection of cholesterol. Analytical imprecision was <10 %CV for 24(S)-, 25-, 27-, 7α-hydroxycholesterol (HC) and cholesterol and ≤15 % for 7-keto-cholesterol. Multiple Sclerosis and other neurological disorder patients had lower 27-hydroxycholesterol levels compared to controls whereas 7α-hydroxycholesterol was lower specifically in Multiple Sclerosis.ConclusionThe method is suitable for measuring plasma oxysterols levels in human health and disease. Analysis of human plasma indicates that the oxysterol, bile acid precursors 7α-hydroxycholesterol and 27-hydroxycholesterol are lower in Multiple Sclerosis and may serve as potential biomarkers of disease.

Cholesterol and lipoprotein metabolism: Early Career Committee contribution.

Cholesterol and lipoprotein metabolism: Early Career Committee contribution.

Rate of steroid double-bond reduction catalysed by the human steroid 5β-reductase (AKR1D1) is sensitive to steroid structure: implications for steroid metabolism and bile acid synthesis.

Human AKR1D1 (steroid 5β-reductase/aldo-keto reductase 1D1) catalyses the stereospecific reduction of double bonds in Δ4-3-oxosteroids, a unique reaction that introduces a 90° bend at the A/B ring fusion to yield 5β-dihydrosteroids. AKR1D1 is the only enzyme capable of steroid 5β-reduction in humans and plays critical physiological roles. In steroid hormone metabolism, AKR1D1 serves mainly to inactivate the major classes of steroid hormones. AKR1D1 also catalyses key steps of the biosynthetic pathway of bile acids, which regulate lipid emulsification and cholesterol homoeostasis. Interestingly, AKR1D1 displayed a 20-fold variation in the kcat values, with steroid hormone substrates (e.g. aldosterone, testosterone and cortisone) having significantly higher kcat values than steroids with longer side chains (e.g. 7α-hydroxycholestenone, a bile acid precursor). Transient kinetic analysis revealed striking variations up to two orders of magnitude in the rate of the chemistry step (kchem), which resulted in different rate determining steps for the fast and slow substrates. By contrast, similar Kd values were observed for representative fast and slow substrates, suggesting similar rates of release for different steroid products. The release of NADP+ was shown to control the overall turnover for fast substrates, but not for slow substrates. Despite having high kchem values with steroid hormones, the kinetic control of AKR1D1 is consistent with the enzyme catalysing the slowest step in the catabolic sequence of steroid hormone transformation in the liver. The inherent slowness of the conversion of the bile acid precursor by AKR1D1 is also indicative of a regulatory role in bile acid synthesis.

PTH-107 A Preliminary Study Of The Effects Of Obeticholic Acid, A Farnesoid X Receptor Agonist, In Patients With Chronic Diarrhoea Secondary To Crohn’s Ileal Disease

IntroductionChronic diarrhoea occurs frequently as a result of excess faecal bile acid (BA) loss. Secondary bile acid diarrhoea (SBAD) is common in Crohn’s disease with ileal inflammation and/or resection. The...

A useful multi-analyte blood test for cerebrotendinous xanthomatosis

ObjectivesCerebrotendinous xanthomatosis (CTX) is a rare genetic disorder of bile acid (BA) synthesis that can cause progressive neurological damage and premature death. Blood (normally serum or plasma) testing for CTX is performed by a small number of specialized laboratories, routinely by gas chromatography–mass spectrometry (GC–MS) measurement of elevated 5α-cholestanol. We report here on a more sensitive biochemical approach to test for CTX particularly useful for confirmation of CTX in the case of a challenging diagnostic sample with 5α-cholestanol that, although elevated, was below the cut-off used for diagnosis of CTX (10μg/mL or 1.0mg/dL). Design and methodsWe have previously described liquid chromatography–electrospray ionization-tandem mass spectrometry (LC–ESI-MS/MS) methodology utilizing keto derivatization to enable the sensitive quantification of plasma ketosterol BA precursors that accumulate in CTX. We have expanded this methodology to perform isotope dilution LC–ESI-MS/MS quantification of a panel of plasma ketosterol BA precursors, with internal standards readily generated using isotopically-enriched derivatization reagent. ResultsQuantification of plasma ketosterol BA precursors (7α-hydroxy-4-cholesten-3-one, 7α,12α-dihydroxy-4-cholesten-3-one and 7α,12α-dihydroxy-5β-cholestan-3-one) in a single LC–ESI/MS/MS test provided better discrimination between a CTX-positive and negative samples analyzed (n=20) than measurement of 5α-cholestanol alone. ConclusionsQuantification of plasma ketosterol BA precursors provides a more sensitive biochemical approach to discriminate between CTX negative and positive samples. A multiplexed LC–ESI-MS/MS test quantifying a panel of plasma ketosterols, with simple sample preparation, rapid analysis time and readily available internal standards, can be performed by most clinical laboratories. Wider availability of testing will benefit those affected with CTX.

Bile acid synthesis precursors in familial combined hyperlipidemia: The oxysterols 24S-hydroxycholesterol and 27-hydroxycholesterol

Familial combined hyperlipidemia (FCHL), the most common inherited disorder of lipid metabolism is characterized by increasing cholesterol synthesis precursors due to hepatic overproduction of cholesterol. The bile acids synthesis pathway has not been previously studied in FCHL. The aim of this work was to study the oxysterol levels which are involved in the bile acids synthesis from cholesterol in FCHL. Clinical parameters and subclinical atherosclerosis were studied in a total of 107 FCHL patients and 126 normolipidemic controls. Non cholesterol sterols (desmosterol and lanosterol) and oxysterols (27-hydroxycholesterol and 24S-hydroxycholesterol) were measured by high performance liquid chromatography tandem mass spectrometry. Desmosterol and lanosterol, markers of cholesterol synthesis, had a positive correlation with BMI and apo B. However, no correlation was found for 24S-hydroxycholesterol and 27-hydroxycholesterol, precursors of bile acids, with these clinical parameters. Only 27-hydroxycholesterol had a positive correlation with apo B, ρ=0.204 (P=0.037). All oxysterol levels were higher in FHCL as compared to normal controls. A total of 59 FCHL subjects (59%) presented values of 24S-hydroxycholesterol above the 95th percentile of this oxysterol in the control population. All oxysterols showed no association with fat mass in contrast with non-cholesterol sterols. FCHL subjects with oxysterol overproduction had less carotid intima media thickness (cIMT), which suggests less atherosclerosis in these subjects. In summary, our data indicate that high oxysterol levels might be good markers of FCHL, unrelated to fat mass, and may exert a protective mechanism for cholesterol accumulation.

Physiological and pathological implications of cholesterol

Cholesterol has evolved to fulfill sophisticated biophysical, cell signaling and endocrine requirements of animal systems. At a cellular level, cholesterol is found in membranes, where it increases both bilayer stiffness and impermeability to water and ions. Furthermore, cholesterol is integrated into specialized lipid-protein membrane microdomains with critical topographical and signaling functions. At an organismal level, cholesterol is the precursor for all steroid hormones, including gluco- and mineralo-corticoids, sex hormones and vitamin D, all of which regulate carbohydrate, sodium, reproductive and bone homeostasis, respectively. This sterol is also the precursor for bile acids, which are important for intestinal absorption of dietary lipids as well as energy and glucose metabolic regulation. Importantly, complex mechanisms maintain cholesterol within physiological ranges and the disregulation of these mechanisms results in embryonic or adult diseases, caused by either excessive or reduced tissue cholesterol levels. The causative role of cholesterol in these diseases has been demonstrated by diverse genetic and pharmacologic animal models that are commented in this review.

A blood test for cerebrotendinous xanthomatosis with potential for disease detection in newborns

Cerebrotendinous xanthomatosis (CTX) is a rare, difficult-to-diagnose genetic disorder of bile acid (BA) synthesis that can cause progressive neurological damage and premature death. Detection of CTX in the newborn period would be beneficial because an effective oral therapy for CTX is available to prevent disease progression. There is no suitable test to screen newborn dried bloodspots (DBS) for CTX. Blood screening for CTX is currently performed by GC-MS measurement of elevated 5α-cholestanol. We present here LC-ESI/MS/MS methodology utilizing keto derivatization with (O-(3-trimethylammonium-propyl) hydroxylamine) reagent to enable sensitive detection of ketosterol BA precursors that accumulate in CTX. The availability of isotopically enriched derivatization reagent allowed ready tagging of ketosterols to generate internal standards for isotope dilution quantification. Ketosterols were quantified and their utility as markers for CTX was compared with 5α-cholestanol. 7α,12α-Dihydroxy-4-cholesten-3-one provided the best discrimination between CTX and unaffected samples. In two CTX, newborn DBS concentrations of this ketosterol (120-214 ng/ml) were ∼10-fold higher than in unaffected newborn DBS (16.4 ± 6.0 ng/ml), such that quantification of this ketosterol provides a test with potential to screen newborn DBS for CTX. Early detection and intervention through newborn screening would greatly benefit those affected with CTX by preventing morbidity and mortality.

Cerebrotendinous xanthomatosis

Cerebrotendinous xanthomatosis (CTX) is a rare neurological disease characterized by accumulation of cholesterol and cholestanol in brain and tendons caused by a mutation in the sterol 27-hydroxylase gene (CYP27A1). The mechanism behind the accumulation of cholestanol in the brain was recently clarified and a role of 27-hydroxycholesterol as a regulator of brain cholesterol homeostasis has been established. There is a significant flux of the bile acid precursor 7α-hydroxy-4-cholesten-3-one across the blood-brain barrier in cy27-/- mice with its subsequent conversion into cholestanol. CTX patients with white matter lesions and vacuolation are described. CYP27A1 was identified as a candidate gene for sporadic amyotrophic lateral sclerosis (ALS). The mechanism behind accumulation of cholestanol in brain and tendons of patients with CTX has been clarified but it is not known why this accumulation is associated with parallel accumulation of cholesterol and formation of xanthomas. Further studies are needed to understand why some patients with CTX develop white matter lesions in the brain. In view of the fact that CTX can present with upper motor neuronal signs it is interesting that CYP27 has been shown to be a candidate gene for sporadic ALS.

Sterol metabolism disorders and neurodevelopment—an update

Cholesterol has numerous quintessential functions in normal cell physiology, as well as in embryonic and postnatal development. It is a major component of cell membranes and myelin, and is a precursor of steroid hormones and bile acids. The development of the blood brain barrier likely around 12-18 weeks of human gestation makes the developing embryonic/fetal brain dependent on endogenous cholesterol synthesis. Known enzyme defects along the cholesterol biosynthetic pathway result in a host of neurodevelopmental and behavioral findings along with CNS structural anomalies. In this article, we review sterol synthesis disorders in the pre- and post-squalene pathway highlighting neurodevelopmental aspects that underlie the clinical presentations and course of Smith-Lemli-Opitz Syndrome (SLOS), mevalonic aciduria (MVA) or the milder version hyper-immunoglobulinemia D and periodic fever syndrome (HIDS), Antley-Bixler syndrome with genital anomalies and disordered steroidogenesis (ABS1), congenital hemidysplasia with icthyosiform nevus and limb defects (CHILD) syndrome, CK syndrome, sterol C4 methyl oxidase (SC4MOL) deficiency, X-linked dominant chondrodysplasia punctata 2(CDPX2)/ Conradi Hunermann syndrome, lathosterolosis and desmosterolosis, We also discuss current controversies and share thoughts on future directions in the field.

Retracted: Advances in the physiological and pathological implications of cholesterol

Cholesterol has evolved to fulfill sophisticated biophysical, cell signalling, and endocrine functions in animal systems. At the cellular level, cholesterol is found in membranes where it increases both bilayer stiffness and impermeability to water and ions. Furthermore, cholesterol is integrated into specialized lipid-protein membrane microdomains with critical topographical and signalling functions. At the organismal level, cholesterol is the precursor of all steroid hormones, including gluco- and mineralo-corticoids, sex hormones, and vitamin D, which regulate carbohydrate, sodium, reproductive, and bone homeostasis, respectively. This sterol is also the immediate precursor of bile acids, which are important for intestinal absorption of dietary lipids as well as energy homeostasis and glucose regulation. Complex mechanisms maintain cholesterol within physiological ranges and the dysregulation of these mechanisms results in embryonic or adult diseases, caused by either excessive or reduced tissue cholesterol levels. The causative role of cholesterol in these conditions has been demonstrated by genetic and pharmacological manipulations in animal models of human disease that are discussed herein. Importantly, the understanding of basic aspects of cholesterol biology has led to the development of high-impact pharmaceutical therapies during the past century. The continuing effort to offer successful treatments for prevalent cholesterol-related diseases, such as atherosclerosis and neurodegenerative disorders, warrants further interdisciplinary research in the coming decades.

The role of cholesterol metabolism and cholesterol transport in carcinogenesis: a review of scientific findings, relevant to future cancer therapeutics

While the unique metabolic activities of malignant tissues as potential targets for cancer therapeutics has been the subject of several recent reviews, the role of cholesterol metabolism in this context is yet to be fully explored. Cholesterol is an essential component of mammalian cell membranes as well as a precursor of bile acids and steroid hormones. The hypothesis that cancer cells need excess cholesterol and intermediates of the cholesterol biosynthesis pathway to maintain a high level of proliferation is well accepted, however the mechanisms by which malignant cells and tissues reprogram cholesterol synthesis, uptake and efflux are yet to be fully elucidated as potential therapeutic targets. High and low density plasma lipoproteins are the likely major suppliers of cholesterol to cancer cells and tumors, potentially via receptor mediated mechanisms. This review is primarily focused on the role(s) of lipoproteins in carcinogenesis, and their future roles as drug delivery vehicles for targeted cancer chemotherapy.

The effects of bovine milk fat on human health

Recent years have witnessed a growing interest in the nutritional value and health benefits of food products of animal origin. Numerous research studies have been undertaken to evaluate the effects of bovine milk, a key dietary component, on human health. Fat is one of the most important components in bovine milk, and its content ranges from 2.8% to 8.1%, subject to the breed of cattle, nutritional aspects, individual characteristics, lactation period, milk production hygiene and season. The aim of this study was to review the latest literature concerning the health effects of components found in bovine milk fat. This paper is a literature review, and it analyzes the composition of bovine milk fat and its effects on human health. The available sources were grouped thematically, and an attempt was made to characterize various milk fat components and their effects on human health. The unique nutritional value of bovine milk can be attributed to the presence of short-chain fatty acids and medium-chain fatty acids which are important sources of energy for the muscles, heart, liver, kidneys, blood platelets and nervous system. They do not pose an obesity risk; they prevent ulcerative colitis, cancer, atherosclerosis and hypertension; they have anti-inflammatory and antibacterial effects, and they boost natural immunity. Milk contains cholesterol, a lipid derivative which stabilizes and stiffens cell membranes, builds the cell cytoskeleton, protects nerve fibers and acts as a precursor of steroid hormones, bile acids and vitamin D 3 . Bovine milk lipids do not exert hypercholesterolemic or atherogenic effects in the human body. A growing tendency to replace animal fats, mainly milk fat, with vegetable fats is a matter of concern.

Five decades with oxysterols

I have been involved in research on oxysterols since 1963 and this review is intended to cover some of the most important aspects of this work. The first project dealed with 7α-hydroxy-4-cholesten-3-one. My successful synthesis of this steroid with high specific radioactivity allowed a demonstration that it is a bile acid precursor. The mechanism of conversion of 7α-hydroxycholesterol into 7α-hydroxy-4-cholesten-3-one was investigated and I concluded that only one enzyme is required and that no isomerase is involved. Accumulation of 7α-hydroxy-4-cholesten-3-one in patients with lack of sterol 27-hydroxylase (Cerebrotendinous xanthomatosis was shown to be an important pathogenetic factor. This disease is characterized by cholestanol-containing xanthomas in tendons and brain and we could show that most of this cholestanol is formed from 7α-hydroxy-4-cholesten-3-one. We also showed that 7α-hydroxy-4-cholesten-3-one passes the blood-brain barrier. In contrast to cholesterol itself, side-chain oxidized oxysterols have a high capacity to pass lipophilic membranes. We demonstrated conversion of cholesterol into 27-hydroxycholesterol to be a significant mechanism for elimination of cholesterol from macrophages. We also showed that conversion of cholesterol into 24S-hydroxycholesterol is important for elimination of cholesterol from the brain. Side-chain oxidized oxysterols have a high capacity to affect critical genes in cholesterol turnover in vitro. Most of the published in vitro experiments with oxysteroids are highly unphysiological, however. Mouse models studied in my laboratory with high or low levels of 27-hydroxycholesterol have little or no disturbances in cholesterol homeostasis. 24S-hydroxycholesterol is an efficient ligand to LXR and suggested to be important for cholesterol homeostasis in the brain. We recently developed a mouse model with markedly increased levels of this oxysterol in circulation and brain. This overexpression had however only a very modest effect on cholesterol turnover. We concluded that oxysterols are not the master regulators of cholesterol homeostasis in vivo suggested previously.

Pleiotropic Functions of the Organic Solute Transporter Ostα-Ostβ

The heteromeric organic solute transporter alpha-beta (Ostα-Ostβ) is expressed at relatively high levels on the basolateral membrane of enterocytes, where it plays a critical role in the intestinal absorption of bile acids and the enterohepatic circulation. However, this transporter is also expressed in nearly all human tissues, including those that are not normally thought to be involved in bile acid homeostasis, indicating that Ostα-Ostβ may have additional roles beyond bile acid transport in these other tissues, or that bile acids and their derivatives are more pervasive than currently envisioned. Emerging data from different laboratories provide support for both of these hypotheses. In particular, recent studies indicate that tissues such as brain and ovary have the capacity to synthesize bile acids or bile acid precursors. In addition, studies examining Ostα-Ostβ substrate specificity have revealed that this transporter can also accept conjugated steroids, including some neurosteroids, and that the transporter is selectively expressed in steroidogenic cells of the brain and adrenal gland, suggesting a novel function for Ostα-Ostβ. The broad tissue expression of Ostα-Ostβ is also consistent with the emerging concept that bile acids and their derivatives act as signaling molecules in diverse tissues. Bile acids activate nuclear receptors such as the farnesoid X receptor (FXR/NR1H4), the pregnane X receptor and the vitamin D receptor, are ligands for a G-protein-coupled bile acid receptor (GPBAR1/TGR5), and can also activate protein kinases A and C as well as mitogen-activated protein kinase pathways. These signaling pathways are present in many tissues and regulate processes such as triglyceride, glucose and energy homeostasis. Note that although FXR and TGR5 are thought to function primarily as bile acid receptors, they are modulated by some other sterols and select lipid metabolites, and are also widely expressed in tissues, indicating a complex interplay among diverse regulatory networks that impact critical cell and organ functions. The present report summarizes the evidence for a pleiotropic role of Ostα-Ostβ in different tissues.

Whole body cholesterol metabolism is impaired in Huntington's disease

We previously reported impaired cholesterol biosynthesis in rodent Huntington Disease (HD) models and HD patients’ fibroblasts and post mortem brains. We also found that plasma levels of 24S-hydroxycholesterol (24OHC), the brain specific elimination product of cholesterol considered a marker of brain cholesterol turnover, were significantly reduced in HD patients at any disease stage. In the present study we analysed by mass spectrometry the fasting plasma levels of cholesterol, its biosynthetic precursors lanosterol and lathosterol, of the whole-body elimination products 27-hydroxycholesterol and of brain 24OHC in a cohort of premanifest and HD patients at different disease stages. We found that the cholesterol precursors lanosterol and lathosterol (both index of whole body cholesterol synthesis), the levels of the bile acid precursor 27-hydroxycholesterol, and of the brain specific 24OHC, were all significantly reduced in manifest HD patients, suggesting that whole-body and brain cholesterol homeostasis are both impaired in HD.