Discrete interplay of gut microbiota L-tryptophan metabolites in host biology and disease.

The gut microbiota and the host maintain a conjoint relationship and together achieve optimal physiology via a multitude of interactive signalling cues. Dietary-derived L-tryptophan (L-trp) is enzymatically metabolized by the resident symbiotic gut microbiota to indole and various indole derivatives. Indole and indole metabolites secreted by the gut bacteria act locally in the intestinal cells as well as distally and modulate tissue-specific functions which are beneficial to the host. Functions attributed to these microbial indole metabolites in the host include regulation of intestinal permeability, immunity and mucosal roles, inflammation, and insulin sensitivity. On the other hand, dysregulation of gut microbiota L-trp metabolism compromises the optimal availability of indole and indole metabolites and can induce the onset of metabolic disorders, inflammation, liver steatosis, and decrease gut barrier integrity. Gut dysbiosis is regarded as one of the prime reasons for this deregulated microbial-derived indole metabolites. A number of indole metabolites from the gut bacteria have been identified recently displaying variable affinity towards xenobiotic nuclear receptors. Microbial metabolite mimicry concept can be used to design and develop novel indole-moiety-containing compounds with higher affinity towards the receptors and efficacy in preclinical studies. Such compounds may serve as therapeutic drugs in clinical trials in the future. In this article, I review L-trp metabolism in the host and gut microbiota and the various physiological functions, patho-physiologies associated with the microbial-released indole metabolites in the host, including the metabolite mimicry-based concept to develop tailored indole-containing novel experimental drugs.

Estrogen-related receptor alpha in select host functions and cancer: new frontiers.

Eukaryotic gene expression is under the tight control of transcription factors, which includes the estrogen-related receptor alpha (ERRα). The endogenous ligand(s) acting as ERRα agonist has not been identified and confirmed. ERRα is a prominent member of the nuclear receptors super-family with major roles in energy metabolism, including immunity, cell growth, proliferation and differentiation and a host of other functions in animals. The actions exerted by ERRα towards gene expression regulation are often in association with other transcriptional factors, receptors and signal mediators. Metabolic regulation by ERRα is known for some time that has tremendous impact on host biology like autophagy, angiogenesis, mitochondrial activity, including lipid metabolism. Cellular metabolism and cancer has intricate relationship. On account of the participation of ERRα in metabolism, it has been implicated in various types of cancer onset and progression. In a number of findings, ERRα has been demonstrated to influence several types of cancers, exhibiting as a negative prognostic marker for many. Such diverse role associated with ERRα is due to its interaction with numerous transcriptional factors and other signalling pathways that culminate in providing optimal gene regulation. These observations points to the crucial regulatory roles of ERRα in health and disease. In this article, some of the new findings on the influence of ERRα in host metabolism and biology including cancer, shall be reviewed that will provide a concise understanding of this receptor.

Human microbial metabolite mimicry as a strategy to expand the chemical space of potential drugs.

The concept of small-molecule mimicry even of weak microbial metabolites present in rodents and humans, as a means to expand drug repertoires, is new. Hitherto, there are few proof-of-concept papers demonstrating utility of this concept. More recently, papers demonstrating mimicry of intestinal microbial metabolites could expand the drug repertoire for diseases such as inflammatory bowel disease (IBD). We opine that, as more functional metabolite-receptor pairings are discovered, small-molecule metabolite mimicry could be a significant effort in drug discovery.

Targeting the pregnane X receptor using microbial metabolite mimicry.

The human PXR (pregnane X receptor), a master regulator of drug metabolism, has essential roles in intestinal homeostasis and abrogating inflammation. Existing PXR ligands have substantial off-target toxicity. Based on prior work that established microbial (indole) metabolites as PXR ligands, we proposed microbial metabolite mimicry as a novel strategy for drug discovery that allows exploiting previously unexplored parts of chemical space. Here, we report functionalized indole derivatives as first-in-class non-cytotoxic PXR agonists as a proof of concept for microbial metabolite mimicry. The lead compound, FKK6 (Felix Kopp Kortagere 6), binds directly to PXR protein in solution, induces PXR-specific target gene expression in cells, human organoids, and mice. FKK6 significantly represses pro-inflammatory cytokine production cells and abrogates inflammation in mice expressing the human PXR gene. The development of FKK6 demonstrates for the first time that microbial metabolite mimicry is a viable strategy for drug discovery and opens the door to underexploited regions of chemical space.

The estrogen-related receptors in metabolism and cancer: newer insights.

The eukaryotic nuclear receptors (NRs) super-family of transcriptional factors include the estrogen-related receptors (ERRs) that have diverse roles in control of cellular energy balance, general metabolism, growth and development, immunity etc. Mouse knock-out models of specific ERR isoforms (ERRα, ERRß and ERRγ) exhibit defects in several phenotypic traits. Newer findings indicate important roles of ERRs in the regulation of brown adipocyte tissue mitochondrial oxidative functions as well as metabolic control in association with hypoxia-inducible factors during cellular hypoxic state. Genes involved in cardiac metabolism is also influenced by ERRα and ERRγ in association with the co-activators PGC-1α and PGC-1ß. On the other hand, ERRs have crucial involvement at the interface of metabolism and diseases such as cancer. Recent findings have implicated ERRα in the progression of tumor and malignancy of the breast, prostate, colon, endometrium etc. In this article, new insights into the regulatory role of ERRs in metabolism and cancer shall be reviewed.

Xenobiotic Receptor-Mediated Regulation of Intestinal Barrier Function and Innate Immunity.

The molecular basis for the regulation of the intestinal barrier is a very fertile research area. A growing body of knowledge supports the targeting of various components of intestinal barrier function as means to treat a variety of diseases, including the inflammatory bowel diseases. Herein, we will summarize the current state of knowledge of key xenobiotic receptor regulators of barrier function, highlighting recent advances, such that the field and its future are succinctly reviewed. We posit that these receptors confer an additional dimension of host-microbe interaction in the gut, by sensing and responding to metabolites released from the symbiotic microbiota, in innate immunity and also in host drug metabolism. The scientific evidence for involvement of the receptors and its molecular basis for the control of barrier function and innate immunity regulation would serve as a rationale towards development of non-toxic probes and ligands as drugs.

Estrogen-related receptor alpha and cancer: axis of evil.

Cancer is perhaps the fastest growing non-communicable disease in the human population worldwide. Although the molecular mechanism of cancer initiation and progression is known to some extent, however, the majority of pathways responsible for its onset, development and progression are largely unknown. Many members of the nuclear receptors (NRs) superfamily of transcriptional factors have key roles in cancer. Estrogen-related receptor alpha (ERRα) is one of the members of the NR superfamily and studies have linked it with a wide variety of cancers. In endocrine-related cancers such as breast cancer, ERRα regulates a number of target genes directing cell proliferation and growth independent of estrogen receptor alpha (ERα). Knockdown of ERRα in a number of cancer tissues and cell lines significantly reduced tumor growth and malignancy indicating dependence on ERRα activity. The pro-angiogenesis factor vascular endothelial growth factor expression has been shown to be regulated by ERRα and has implications in several types of cancer. The effect of ERRα on cancers seems to be multipronged via regulation of cell cycle regulators, osteopontin, hypoxia inducible factor-1 as well as several energy metabolism genes that are part of glycolysis, TCA cycle, lipogenesis, etc., providing a metabolic twist to cancer. In this article, the action of ERRα on various types of cancers including new developments in this field shall be reviewed.

The orphan estrogen-related receptor alpha and metabolic regulation: new frontiers.

Metabolic homeostasis during long-term adaptation in animals is primarily achieved by controlling the expression of metabolic genes by a plethora of cellular transcription factors. The nuclear receptor (NR) superfamily in eukaryotes is an assembly of diverse receptors working as transcriptional regulators of multiple genes. The orphan estrogen-related receptor alpha (ERRα) is one such receptor of the NR superfamily with significant influence on numerous metabolic and other genes. Although it is presently unknown as to which endogenous hormones or ligands activate ERRα, nevertheless it regulates a host of genes whose products participate in various metabolic pathways. Studies over the years show new and interesting data that add to the growing knowledge on ERRα and metabolic regulation. For instance, novel findings indicate existence of mTOR/ERRα regulatory axis and also that ERRα control PGC-1α expression which potentially have significant impact on cellular metabolism. Data show that ERRα exerts its metabolic control by regulating the expression of SIRT5 that influences oxygen consumption and ATP generation. Moreover, ERRα has a role in creatine and lactate uptake in skeletal muscle which is important towards energy generation and contraction. This review is focused on the new insights gained into ERRα regulation of metabolism, networks and pathways that have important consequences in maintaining metabolic homeostasis including development of cancer.

The NR4A orphan nuclear receptors: mediators in metabolism and diseases.

The NR4A subfamily is orphan nuclear receptors that belong to the larger nuclear receptors (NRs) superfamily of eukaryotic transcription factors. The NR4A subfamily includes three members, namely Nur77 (NR4A1), Nurr1 (NR4A2) and Nor1 (NR4A3) which are gene regulators and participate in diverse biological functions. Though the ligands for these receptors are presently unidentified, they are thought to be constitutively active. NR4A acts as molecular switches in gene regulation and their action is increasingly seen to be modulated by complex network of cellular signaling pathways. Members of the NR4A are expressed in tissue-specific fashion which indicates their selective control of various biological processes. Data reveal a host of functions governed by the NR4A subfamily members including general metabolism, immunity, cellular stress, memory, insulin sensitivity and cardiac homeostasis by regulating specific target genes whose products participates in such processes. Moreover, these receptors have a role in the onset and progression of various diseases such as various types of cancer, inflammation, atherosclerosis and obesity. In this review, a concise overview of the current understanding of the important metabolic roles governed by NR4A members including their participation in a number of diseases shall be provided.

Estrogen-related receptor alpha and mitochondria: tale of the titans.

The estrogen-related receptor alpha (ERRα) governs multitude of biological functions by working as specific transcriptional regulator in animals. Over the past few years, one aspect of ERRα that has seen optimal progress is its control over the mitochondrial physiology. The ERRα not only regulates an array of nuclear genes devoted to mitochondrial functions but also numerous mitochondrial DNA genes that ultimately culminates into this organelle's homeostasis. In fact, ERRα expression is correlated with genes whose functional products are part of the mitochondrial physiology. Studies have indicated that nearly half of the proteins encoded by the mitochondrial genome are regulated by ERRα. Moreover, ERRα controls vital mitochondrial processes such as oxidative metabolism through a network of protein kinases and by regulating the expression of sirtuins like Sirt3. Furthermore, new findings also show that ERRα regulate mitochondrial biogenesis in association with PGC family co-activators such as PGC-1-related co-activator and PGC-1ß and also via cross-talk with mitogen-activated protein kinase kinases and PI3K/(AKT) signaling. The current understanding of the pathways and networks shows strong influence of ERRα in coordinating mitochondrial physiology. This review focuses on the new advances made in understanding the complex and important interface between ERRα and mitochondrial physiology.

The orphan nuclear receptors in cancer and diabetes.

The orphan nuclear receptors (ONRs) are a vital class of transcriptional regulators belonging to the larger nuclear receptors (NRs) superfamily in higher eukaryotes. As a result of non-identification of endogenous physiological ligands for this class of NRs, they are designated as "orphans". The ONRs on receiving appropriate signals translate into specific gene regulation. Elaborate studies on the ONRs in the past two decades have revealed crucial biological functions controlled by them relating to general metabolism, immunity, organogenesis, angiogenesis, growth and development, and numerous other tissue physiologies. Over the years, many of the ONRs have been studied for their participatory role in human health and disease. Results obtained are encouraging and interesting and shows a number of ONRs does modulate several patho-physiological conditions such as cancer and diabetes. This review discusses the current status about the interplay between select ONRs in cancer and diabetes.

The interplay between retinoic acid receptor-related orphan receptors and human diseases.

The retinoic acid receptor-related orphan receptors (RORs) are an important subfamily of transcriptional regulators of the nuclear receptors superfamily. Their discovery over a decade ago by gene cloning strategy have revealed three major isoforms of these orphan receptors in animals. Generation and analyses of isoform-specific ROR null mice have provided revealed-vital roles for the RORs in animals. The RORs undoubtedly participate in a host of biological functions such a metabolism, immunity, development and differentiation, angiogenesis, circadian clock, xenobiotic/drug metabolism and other tissue physiologies for optimal animal survival. Moreover, intense work in the last one decade also revealed a host of human diseases being modulated by the RORs. A number of diseases, such as cancer, autoimmune diseases, inflammation, osteoporosis, metabolic syndrome etc., strongly support the involvement of RORs in their onset and progression. By involving in such diseases, the RORs are indeed a critical factor for optimal cell function and are being intensely investigated as novel targets for drug interventions in the treatment of various diseases. This review focuses on the current knowledge and status about RORs in a number of human disease conditions.

The estrogen-related receptors: orphans orchestrating myriad functions.

Coordinated and tight regulation of gene expression in metazoans is essential for cellular homeostasis and functions. Tissue- and cell-specific regulatory factors are indispensable and a wide variety of them exist to regulate genes. A family of transcriptional factors was identified in the past two decades through gene cloning studies and was informally referred as "orphan receptors", as appropriate endogenous ligands for such receptors were unknown. One of the subclasses of such receptors is known as the estrogen-related receptors (ERRs), which include three isoforms, namely ERRα, ERRß and ERRγ. Over the past one decade, unprecedented knowledge about the ERRs biology has been generated, indicating their vital roles in various metabolic and physiological activities in animals. The ERRs cellular action is largely attributed to its interaction with a wide variety of other nuclear receptors, including some orphan nuclear receptors, and thereby can modulate diverse array of genes involved in metabolism and animal physiology. Studies using genome-wide location analyses, microarray and functional genomics, including ERR-specific null mice have revealed a number of pathways controlled by the ERRs. In this context, new and recent information on the biological functions of ERRs are being reviewed.

The mammalian orphan nuclear receptors: orphans as cellular guardians.

In classical endocrinology, receptors are molecules that bind a hormone or a ligand to transduce signal within a target cell. Later, however, many intracellular receptors have been discovered in mammals, which have not been shown to bind endogenous ligands and are now are referred as "orphan receptors." The orphan receptors share high degree of structural and functional homology with the classical nuclear receptors (NRs) and are now part of the NR superfamily and therefore referred as orphan nuclear receptors (ONRs). Interestingly, however, ONR members are not evolutionarily or functionally linked and they form a highly diverse group within the NR superfamily. In mammals, ONRs exhibit great functional diversity and majority of them are expressed in a tissue-specific fashion. In the past one decade, functional studies have revealed that they are mediators of multitude of crucial metabolic, developmental, reproductive, and immunological functions in mammals. Emerging studies also indicate the role of ONRs in the onset of several complex human diseases and hence they may be potential candidates for therapeutic drug targeting in the future.

The estrogen-related receptor alpha: the oldest, yet an energetic orphan with robust biological functions.

The estrogen-related receptor alpha (ERRalpha) is an orphan nuclear receptor (ONR) that by binding to DNA sites controls gene expression in association with coactivators and corepressors. ERRalpha was the first ONR to be identified; however, its natural endogenous ligand(s) is still unknown. ERRalpha by acting as a transcription factor has been shown to regulate a large array of genes, thereby controlling numerous metabolic pathways and other biological functions in animals. Of late, the expression of ERRalpha has been detected in several tissues, including those with high metabolic activities and energy demand. Presently, the control of energy balance by ERRalpha seems to be its prime role. The nonavailability of endogenous ligand for ERRalpha has not impeded the study of its functions. In fact, most of the present knowledge of the biological roles of ERRalpha has evolved from in-depth biochemical, overexpression, genomic, including functional genomics studies, and also through the generation of intact ERRalpha knockout (null) mice. Interestingly, over the past few years, growing evidence suggests interplay between ERRalpha and various human metabolic diseases such as diabetes, obesity, and heart disease. Also, there are strong indications of the involvement of ERRalpha in cancer initiation and progression. Interestingly, this makes ERRalpha a suitable, direct target for pharmacological intervention in treating such diseases. This review focuses on the overall developments and recent advances in understanding the role of ERRalpha in metabolism and other biological functions, including its role in human diseases.

Long-term caloric restriction up-regulates PPAR gamma co-activator 1 alpha (PGC-1alpha) expression in mice.

The peroxisome proliferator-activated receptor (PPAR) gamma co-activator 1 alpha (PGC-1alpha), a signal-sensing transcriptional co-activator in association with many nuclear receptors regulates various genes that control energy balance in animals. In this study, the effect of long-term caloric restriction (CR) (alternate days of fasting for 3 months) on the expression of PGC-1alpha protein in various tissues was investigated in mice. Western blot analyses showed positive immunoreactive PGC-1alpha (approximately 92 kDa) signal from various tissues. Heart, kidney and skeletal muscles expressed significant levels of PGC-1alpha, while a comparatively lower level was detected in the liver, small intestine and brain. The expression of PGC-1alpha was the highest and lowest in the heart and liver respectively. CR mice exhibited a significant increase in PGC-1alpha level in the heart (5.13-fold), kidney (3.57-fold), skeletal muscle (3.02-fold), liver (2.60-fold), small intestine (2.45-fold) and brain (2.05-fold), compared to normal (ad libitum) fed. The elevation in PGC-1alpha level, especially in highly oxidative tissues such as heart, kidney and skeletal muscle of CR mice might synergistically up-regulate genes that require PGC-1alpha co-activation. Taken together, the up-regulation of PGC-1alpha expression might potentially support optimal energy metabolism and biochemical adaptation, necessary for maintaining energy homeostasis during long-term CR.

Moderately high altitude habitation modulates lipid profile and alkaline phosphatase activity in aged Khasis of Meghalaya.

The indigenous Khasis inhabit different geographical and climatic locations of Meghalaya. In this study, we intended to find out whether habitation in moderately high altitude place has any effect on the lipid and liver profile amongst the aged Khasis. The level of various serum parameters under lipid and liver profile were analyzed and compared from aged (65-70 years) male Khasi residents of moderately high (Shillong city) and low (Byrnihat) altitude places. Results obtained from the lipid profile data show decreased total serum cholesterol (29%), triglyceride (27%) and LDL-cholesterol (42%) level in the old Khasis of Shillong compared to Byrnihat. Furthermore, the alkaline phosphatase activity was significantly raised (47%) in the old Khasis from Shillong as against Byrnihat. The decreased level of total cholesterol, triglyceride and LDL-cholesterol in old Khasis from Shillong may be due to living and acclimatization in high altitude with low annual temperature. Moderately high elevation could have acted as a stressor, thereby reducing the level of serum cholesterol, triglyceride and LDL-cholesterol, which may put them at a lower risk of cardiovascular diseases. In comparison, old residents of Byrnihat with high cholesterol, triglyceride and LDL-cholesterol levels may elevate their risk of coronary complications. The raised alkaline phosphatase activity amongst the old Khasis of Shillong could be due to increased bone and/or intestinal turnover as a result of living in high altitude, which, however, may elevate the risk of osteoporosis. Taken together, we conclude that low cholesterol, triglyceride and LDL-cholesterol levels, accompanied with high alkaline phosphatase activity amongst the old Khasis of Shillong, could be due to the influence of high altitude and mild climatic conditions that prevails.

Up-regulation of orphan nuclear estrogen-related receptor alpha expression during long-term caloric restriction in mice.

The estrogen-related receptor alpha (ERRalpha) is an orphan receptor belonging to the nuclear receptor superfamily that regulates a number of target genes encoding enzymes that participate in various metabolic pathways involved in maintaining energy balance in animals. In this study, whether long-term caloric restriction (alternate days of fasting for 3 months) in mice modulates the expression of ERRalpha in various tissues was investigated. Western blot analyses showed positive immunoreactive ERRalpha protein (53 kDa) band in various mice tissue extracts, though at varying levels. Heart, kidney, and skeletal muscles expressed significant levels of ERRalpha, with a comparatively lower level detected in the intestine, brain, and liver. Cardiac ERRalpha expression was the highest, with the least detected in the liver. Caloric restricted mice exhibited a significant increase in ERRalpha level in the heart (5.45-fold), kidney (3.70-fold), skeletal muscle (3.0-fold), small intestine (2.72-fold), and liver (2.44-fold) extracts as compared to ad libitum fed. However, caloric restriction could not evoke any detectable receptor level change in the brain. Notably, the highest ERRalpha up-regulation was detected in the heart. This up-regulation in ERRalpha level especially in highly oxidative tissues such as heart, kidney, small intestine, and skeletal muscle of caloric restricted mice may be helpful in modulating ERRalpha responsive genes that participates in maintaining energy balance. This may potentially strengthen the metabolic and biochemical adaptation in such tissues, which is necessary for animal survival under long-term caloric restriction.

Assessing the estrogenicity of environmental chemicals with a stably transfected lactoferrin gene promoter reporter in HeLa cells.

Lactoferrin is an important marker protein of the estrogens. In mice, lactoferrin expression is stimulated in the uterus by ligand-bound estrogen receptors (ERs). With this study we aimed to evaluate the effect of different environmental estrogenic chemicals on the mouse lactoferrin gene expression in a cell-based assay. We constructed a reporter that contains the firefly luciferase gene under hormone-inducible control of a 1.1kbp fragment of the mouse lactoferrin gene promoter. In an attempt to study the promoter regulation in a chromatin context, we stably transfected the construct (pGL3-mLF-Luc) into HeLa cells, and a stable clone (HeLa-mLF-Luc) incorporating the construct was subsequently generated. Transient transfection of HeLa-mLF-Luc cells with ERα and ERß expression plasmids showed that both 17ß-estradiol (E2) and diethylstilbestrol (DES) at 10(-7)M significantly increased luciferase expression via ERα and ERß. Xenoestrogens such as bisphenol A, 4-octylphenol, 4-nonylphenol and the phytoestrogen genistein when used at increasing concentrations (10(-8) to 10(-5)M) revealed varying magnitudes of activation (1.96-8-fold). The environmental estrogens showed similar magnitudes of luciferase induction when acting through ERα and ERß-mediated pathways. Also, in the absence of ERs, the xenoestrogens could not induce luciferase expression thereby reflecting receptor dependency. Taken together, the results indicate a significant responsiveness of the stably transfected mouse lactoferrin promoter to endogenous estrogen and environmental estrogenic compounds through ERs. This cell-based transcription assay system may be useful in understanding the susceptibility of estrogen target gene expression by these chemicals at the chromatin level.