See discussions, stats, and author profiles for this publication at: https://www.researchgate.net/publication/24409642 Minireview: evolution of NURSA, the Nuclear Receptor Signaling Atlas Article in Molecular Endocrinology · June 2009 DOI: 10.1210/me.2009-0135 · Source: PubMed CITATIONS READS 72 69 17 authors, including: Austin Cooney Michael Downes 145 PUBLICATIONS 6,899 CITATIONS 159 PUBLICATIONS 12,352 CITATIONS University of Texas Austin Dell Medical School SEE PROFILE Salk Institute SEE PROFILE Mitchell A Lazar Ronald N Margolis 362 PUBLICATIONS 40,843 CITATIONS 46 PUBLICATIONS 792 CITATIONS University of Pennsylvania SEE PROFILE National Institutes of Health SEE PROFILE Some of the authors of this publication are also working on these related projects: Generation of functional cardiomyocytes View project All content following this page was uploaded by Austin Cooney on 20 December 2016. The user has requested enhancement of the downloaded file. MINIREVIEW Minireview: Evolution of NURSA, the Nuclear Receptor Signaling Atlas Neil J. McKenna, Austin J. Cooney, Francesco J. DeMayo, Michael Downes, Christopher K. Glass, Rainer B. Lanz, Mitchell A. Lazar, David J. Mangelsdorf, David D. Moore, Jun Qin, David L. Steffen, Ming-Jer Tsai, Sophia Y. Tsai, Ruth Yu, Ronald N. Margolis, Ronald M. Evans, and Bert W. O’Malley Departments of Molecular and Cellular Biology (N.J.M., A.J.C., F.J.D., R.B.L., D.D.M., M.-J.T., S.Y.T., B.W.O.) and Biochemistry (J.Q.) and The Bioinformatics Research Center (D.L.S.), Baylor College of Medicine, Houston, Texas 77030; Cellular and Molecular Medicine (C.K.G.), University of California San Diego, San Diego California 92093; Division of Endocrinology, Diabetes, and Metabolism (M.A.L.), University of Pennsylvania, Philadelphia, Pennsylvania 19104; Department of Pharmacology (D.J.M.), University of Texas Southwestern Medical Center, Dallas Texas 75390; Division of Diabetes, Endocrinology, and Metabolic Diseases (R.N.M.), National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20892; and Gene Expression Laboratory (M.D., R.Y., R.M.E.), The Salk Institute, La Jolla, California 92037 Nuclear receptors and coregulators are multifaceted players in normal metabolic and homeostatic processes in addition to a variety of disease states including cancer, inflammation, diabetes, obesity, and atherosclerosis. Over the past 7 yr, the Nuclear Receptor Signaling Atlas (NURSA) research consortium has worked toward establishing a discovery-driven platform designed to address key questions concerning the expression, organization, and function of these molecules in a variety of experimental model systems. By applying powerful technologies such as quantitative PCR, high-throughput mass spectrometry, and embryonic stem cell manipulation, we are pursuing these questions in a series of transcriptomics-, proteomics-, and metabolomics-based research projects and resources. The consortium’s web site (www.nursa.org) integrates NURSA datasets and existing public datasets with the ultimate goal of furnishing the bench scientist with a comprehensive framework for hypothesis generation, modeling, and testing. We place a strong emphasis on community input into the development of this resource and to this end have published datasets from academic and industrial laboratories, established strategic alliances with Endocrine Society journals, and are developing tools to allow web site users to act as data curators. With the ongoing support of the nuclear receptor and coregulator signaling communities, we believe that NURSA can make a lasting contribution to research in this dynamic field. (Molecular Endocrinology 23: 740 –746, 2009) T he success of the Human Genome Project and its offshoot genomic and proteomic projects such as ENCODE and HAPMAP represent the fruits of large-scale collaborative efforts and contributions from multiple disciplines (1– 4). Organizing and mining the information contained in genome sequences and their expressed transcriptomes and proteomes has profound implications for our understanding of physiology and pathology, allowing for the development and characterization of models for disease and, ultimately, applications in the clinic. Following on the success of these broad initiatives is a new generation of more tightly focused team science projects encompassing spe- cific groups of molecules, tissues, or disease states. One such project is the Nuclear Receptor Signaling Atlas (NURSA), a consortium of U.S. investigators conceived in 2002 as a vehicle for developing a global approach to investigating nuclear receptor (NR) and coregulator function in physiology and disease (5). ISSN Print 0888-8809 ISSN Online 1944-9917 Printed in U.S.A. Copyright © 2009 by The Endocrine Society doi: 10.1210/me.2009-0135 Received March 24, 2009. Accepted April 27, 2009. First Published Online May 7, 2009 Abbreviations: ERR, Estrogen-related receptor; ES, embryonic stem; GR, glucocorticoid receptor; NR, nuclear receptor; NURSA, Nuclear Receptor Signaling Atlas; PPAR, peroxisome proliferator-activated receptor; SRC, steroid receptor coactivator. 740 mend.endojournals.org NRs and Coregulators The NR superfamily represents a highly conserved, widely expressed group of ligand-regulated transcription factors that function in response to dietary and endocrine signals, with roles Mol Endocrinol, June 2009, 23(6):740 –746 Mol Endocrinol, June 2009, 23(6):740 –746 in development, cancer, toxicology, reproduction, and metabolism (6). The NR superfamily in mammals is sufficiently focused (49 known receptors in human, 48 in mouse) to make it possible to explore in greater depth the roles played by these transcription factors, particularly in the regulation of metabolism. When cDNAs encoding members of this family were first cloned and sequenced over 20 yr ago, several common structural motifs that linked them together helped to define their mechanism. The subsequent discovery and characterization over the last decade or so of a large group of molecules, the coregulators, which are required for efficient function of NRs, has sketched an additional level of functional flexibility, and complexity, in NR signaling pathways (7). Functioning coordinately in modular complexes in the nucleus, NRs and their ligands and coregulators serve to repress or activate the expression of target gene networks. Roles in disease for NRs are well documented in the literature, with the roles of estrogen receptor-␣ in breast cancer and androgen receptor in prostate cancer being among the most well known. Moreover, the therapeutic impact of NR ligands is evident in the treatment of a spectrum of disease states, including the estrogen receptor-␣ partial agonist tamoxifen in breast cancer, glucocorticoid receptor (GR)agonists in inflammation, vitamin D receptor agonists in rickets, and peroxisome proliferator-activated receptor (PPAR) agonists in the amelioration of conditions associated with the metabolic syndrome. Although coregulators are at an earlier stage of their characterization as a group compared with NRs, both animal models and clinical studies have implicated them in numerous pathological conditions (8). For example, many coregulators are overexpressed or underexpressed in tumors of the prostate, brain, blood, and breast. In addition, animal models suggest roles for PGC1␣/PPARGC1A in susceptibility to heart failure, for RIP140/NRIP1 in defects in adipocyte differentiation, and for SRC-1/NCOA1 in defects in hepatic function (see Fig. 2 and also the Diseases and Phenotypes module of the specific NURSA Coregulator Pages for references; www.nursa.org). Given this weight of evidence then, coregulators, like NRs, appear to hold tremendous potential as therapeutic leverage points in human disease. NURSA The translational significance with which NRs and coregulators are invested is evinced by the fact that over both phases of its existence (Phase I, 2002–2007, and Phase II, 2007–2012; see Table 1), the consortium has drawn support from five different National Institutes of Health institutes (National Institute of Diabetes and Digestive and Kidney Diseases, National Cancer Institute, National Institute on Aging, National Heart, Lung, and Blood Institute, and National Institute of Environmental Health Sciences). It is well established that NRs and coregulators participate as components in a signaling continuum in the regulation of gene expression and cellular function. As a practical matter, however, given the diverse expertise and focused interests of the investigators involved, it was decided to pursue these parallel research goals in a separate but intertwined fashion, borrowing expertise and exploiting areas of synergy as the mend.endojournals.org 741 TABLE 1. Organization of projects and shared resources in Phase II of NURSA Laboratory Research projects Strand A: genomics 1) NR functional profiling in health and disease 2) Expression profiling in physiological and pathophysiological processes 3) NR coregulator functional pathology in metabolic disease Strand B: proteomics 1) Generation of ES cells to tissuespecifically overexpress NRs and coregulators in vivo 2) Biotin-tagged NRs and coregulators for proteomic and genomic analyses Research resources 1) Proteomic profiling of coregulator complexes 2) Gene profiling 3) Bioinformatics and web site Evans Mangelsdorf/Moore Lazar Tsai/Tsai/DeMayo Glass O’Malley/Qin Evans McKenna projects progressed. Accordingly, research projects and resources were organized into two broad areas, namely 1) functional and expression profiling of NRs in health and disease and 2) proteomic characterization of coregulator complexes (Table 1). In parallel with these, a bioinformatics and web group was established to develop a database for distributing consortium datasets and integrating these with existing resources in the field to create a comprehensive reference resource for the community. Expression Profiling and Functional Genomics of NRs Although much has been learned about the physiology of certain receptors in a limited number of well-studied target tissues, surprisingly little is known about NR function in the majority of tissues throughout the body. This raised the question of whether an understanding of this network could be reached by studying the expression of the NR superfamily as a whole using a systems biology approach. Given that physiology is a process occurring across both spatial and temporal coordinates, we anticipated that this expression blueprint would be expected to have both an anatomical and circadian rhythm component. In two Phase I studies from the Evans and Mangelsdorf laboratories, a highthroughput QPCR platform was used to interrogate the anatomical (9) and circadian (10) expression profiles of the NR superfamily as a means of probing these questions in depth. The quantitative power, reproducibility, and integrity of the QPCR dataset provide distinct advantages of this technique over other high-throughput profiling methods. Direct comparison of this dataset with other databases (e.g. SymAtlas and Mouse Gene Prediction Database) revealed numerous quantitative and qualitative discrepancies, including a number of genes that were not represented at all. Accordingly, the present dataset provides an unparalleled molecular fingerprint of NRs that should provide an important resource tool to the scientific community. 742 McKenna et al. Minireview Mol Endocrinol, June 2009, 23(6):740 –746 come into play at critical junctures to orchestrate the expression of the specific gene networks that drive these systems. Building on the success of the QPCR platform in the first phase, Phase II of NURSA has seen its application to a variety of translational projects. In the Evans (Table 1, genomics strand, project 1) and Mangelsdorf/Moore projects (Table 1, genomics strand, project 2), samples are being profiled from human patients with metabolic disorders, cardiovascular disease, environmental toxin exposure, and cancer. In addition, an array of related mouse models are being screened to investigate potential changes in receptor expression in response to dietary changes associated with obesity, atherosclerosis, hepatic steatosis, and insulin resistance. Proteomic Organization of Coregulators With one or two exceptions, evidence collected by the NURSA Consortium indicates that the tissue expression patterns of coregulators are relatively unremarkable compared with the striking selectivity characteristic of the NRs (14). The key to their function rather may lie in their organization into higher-order steady-state cellular complexes (15), whose dynamic composition and contextual posttranslational modifications appear to be organically linked to the patterns of gene expression directed by NRs and their cognate ligands (16). Accordingly, the Phase I O’Malley and Lazar projects and the current O’Malley and Qin proteomics resource (Table 1, research resource 1) made their long-term goal the characterization of coregulator complexes using a combination of native immunoprecipitation and mass spectrometric methods; the native HeLa cell complexes formed by all NR coregulators are classified by NURSA’s Bioinformatics Resource and comprise a total of over 350 coregulators. Initial efforts were focused on establishing optimal conditions for extract preparation and immunoprecipitation and resulted in the publication of a set of pilot coregulator complexes, including those formed members of the steroid receptor coactivator (SRC)/p160 family, CBP/CREBBP, p300, and SMRT/ NCOR2 (17, 18). Further refinement of the platform was achieved by introducing a two-tier detergent step and reciprocal IP of interacting proteins identified in the initial immunoprecipitation. These additional steps combine to identify false positives and to increase confidence in the true interacting proteins (or co-coregulators) in each complex. We have termed this approach network analysis proteomics because it helps to define the protein interaction network of the original antigen. To date, this strategy has identified and organized 1500 coregulator interactants for 1417 core coregulator proteins (Lanz, R., per- FIG. 1. The NR ring of physiology. [Reprinted from Cell, vol. 126, A. L. Bookout et al., Anatomical profiling of nuclear receptor expression reveals a hierarchical transcriptional network, pages 789 –799. ©2006 with permission from Elsevier (9).] The predictive power of the QPCR data set can be seen in an unbiased clustering analysis of the NR expression profile, which was accomplished by a comprehensive evaluation of the entire NR superfamily in virtually every tissue system (9). Within this ring of NR physiology (Fig. 1), a number of interesting receptor associations becomes evident. Perhaps the most interesting discovery is the inextricable link between NRs and three signature characteristics of animal physiology: reproduction, development, and metabolism. In addition, another prevailing theme revealed by this analysis is the strong nexus that exists between the modulation of nutrient metabolism and immune function, as exemplified by the dual roles of at least 15 different receptors [farnesoid X receptor (FXR), GR, liver X receptors (LXRs), PPARs, retinoic acid receptor-related orphan receptors (RORs), retinoid X receptors (RXRs), and the NR4A subfamily] in these two physiological processes. As just one example of the predictive power of this analysis, we note that the identification of GR, liver X receptor-␣, and PPAR␥ as a distinct group (cluster IIC) has been recently validated by the observation that these receptors function in a combinatorial manner to regulate the evolution of host immune responses (11). Taken together, these data suggest that although individual receptor function is important, they also cluster as part of higher-order regulatory networks. This initial organism-wide approach to systematically documenting receptor expression was followed by a series of more focused studies in specific physiological and pathophysiological contexts, such as adipogenesis (12) and macrophage activation (13). These studies once more afforded an insight into the specific clades of receptors that Mol Endocrinol, June 2009, 23(6):740 –746 sonal communication), which gives some flavor of the dynamic spatiotemporal organization of these complexes and the novel regulatory mechanisms in NR-mediated signaling pathways that they presage. Coregulator complex datasets are released on the public NURSA web site, collectively constituting a unique resource for hypothesis generation and validation by members of the community. The value of this resource can be illustrated with reference to two case studies. Meng et al. (19) used the NURSA coregulator complex datasets to identify PCBP3, as an interacting partner of CAPER/RBM39 (20), allowing them to formulate and test a hypothetical role for PCBP1 in splicing. In another study, Kittler et al. (21) carried out a genome-scale RNA interference screen for human genes important for cell division and identifying SMRT/NCOR2 as one such gene. Citing the presence in the NURSA SMRT complex of TBL1X and MLL5 (22), also present in the histone deacetylaserecruiting SET complex, they proposed a role for a human SET3/SMRT complex in the regulation of genes involved in cytokinesis. Pilot and Collaborative Bridging Projects The Phase I Pilot and Feasibility Projects and the Phase II Collaborative Bridging Projects, although differently named, are predicated upon a common goal: to encourage and foster scientific liaison between the consortium and community investigators with interests and expertise that are complementary to its overall research mission. Broadly speaking, both programs were intended to allow external investigators, through an open, competitive review process, to harness the collective intellectual and infrastructural capacity of the Atlas to catalyze the research output of the consortium as well as to ultimately position them to branch off as independently funded research projects. Examples of these outreach projects are varied. Using a model organism, Caenorhabditis elegans, to test the concept, a pilot project led by Adam Antebi helped to identify natural steroidal hormones in the worm where none before had been seen (23) and to demonstrate the evolutionary conservation of the roles played by steroidal ligands in the regulation of homeostasis. This type of approach may ultimately prove useful for the identification of natural ligands for many orphan NRs. Chemical biology approaches have helped to define ligands for NRs to catalyze functions for, for example, constitutive androstane receptor (24) and to demonstrate the differences in response of constitutive androstane receptor in mouse and human as well as to identify small-molecule agonists for the estrogenrelated receptors (ERRs), with selectivity for the ERR␤ and ERR␥ forms (25). In another example of the variety of NURSA outreach projects, the power of small-molecule agonists/antagonists for NRs has been demonstrated using the compound fexaramine, which was used to help define structural and functional properties of farnesoid X receptor (26). Finally, structural studies carried out in a pilot project led by mend.endojournals.org 743 Eric Xu have also helped to identify putative endogenous ligands for orphan NRs. Community Research Tools To complement the high-content datasets generated by the NURSA research projects, the second phase of NURSA has placed an increased emphasis on generating tools to facilitate research strategies in the wider community. In the genomics strand, the Evans project (Table 1, genomics strand, project 1) includes a component for designing and validating a lentiviralbased murine NR-specific short hairpin RNA library for knockdown of NRs in cells. Extending this approach to tissues, the Lazar genomics project (Table 1, genomics strand, project 3) is pursuing a strategy for tissue-specific knockdown of coregulators by delivery of lentiviral short hairpin RNAs, using the liver as a proof-of-principle tissue for in vivo depletion of the coregulators NCoR, SMRT/NCOR2, and RIP140/NRIP1. Determining the genuine target genes of NRs and coregulators in native chromatin contexts using immunoprecipitationbased techniques is currently an area of intense inquiry in the field. Despite substantial efforts by academic laboratories and antibody companies, the quality of antibodies for many receptors and coregulators often remains a limiting factor in achieving the levels of enrichment required for robust identification of genomic binding sites for these molecules. Although development of antibodies to endogenous proteins will remain an important goal, the Glass proteomics project (Table 1, proteomics strand, project 2) is providing an immediate and versatile alternative by developing and validating vectors that allow for regulated expression of biotin-tagged NRs and coregulators in mammalian cells. These vectors take advantage of the extremely high affinity of the biotin-streptavidin interaction to overcome the inherent limitations of immune-based approaches. The concept of a posttranscriptional code by which coregulator action is in turn regulated has been well characterized in vitro (27), but less well understood is the importance of these modifications in vivo. The O’Malley and Qin proteomics resource (Table 1, research resource 1) contains a component that leverages embryonic stem (ES) cell technology developed by Austin Cooney to systematically define the contribution of specific posttranslationally modified residues to coregulator function in living cells, using the SRC/p160 family as proof-of-concept. ES cell lines containing these mutated forms of the coactivators will be made available to the community. We anticipate that a systematic, nonbiased approach such as this will yield insights into the specific combinations of posttranslational modifications required to drive receptor-, promoter-, and tissuespecific regulation of gene networks in vivo. Although loss-of-function mutations in NRs and coregulators are widely characterized in disease states, there is ample evidence from clinical studies that gain-of-function (through overexpression or gene amplification) of these molecules is a recurring motif in the etiology of a spectrum of pathologies (see Diseases and Phenotypes module of the NURSA Molecule Pages and Fig. 2). With this in mind, the Tsai proteomics project (Ta- 744 McKenna et al. Minireview Mol Endocrinol, June 2009, 23(6):740 –746 which distills into a single menu-driven user interface: data from the NURSA bench projects; content from the leading biological databases, incorporating information relevant on orthologs, datasets, reagents, interactions, diseases and phenotypes; and content curated by members of the NURSA Bioinformatics Resource itself, including systematic organization and meta-analysis of public expression microarray datasets (29), and what we believe to be the first public NR ligand database, eventually comprising information on nearly 250 natural and synthetic ligands. By inviting industry experts in the curation of this database, including those from GSK, Wyeth and Eli Lilly, NURSA is demonstrating its unique role as an information hub for all sections of the research community. Recently revised molecule descriptions have facilitated free text searching for specific content. Through the Molecule Pages it is possible for a user to mine an enormous amount of data - from NURSA-generated datasets and reagents to community-contributed datasets to curated literature archives—to achieve a greater depth of understanding of the complexities of NR and coregulator signaling in health and disease. Outreach FIG. 2. Translational potential of selected NR coregulators in metabolism and cancer. Data are compiled from Ref. 8. The metabolism section identifies phenotypes arising from null deletion of genes in animal models, unless otherwise noted. A comprehensive resource for the role of nuclear receptors and nuclear receptor coregulators in disease can be found in the Molecule Pages Diseases and Phenotypes module on the NURSA web site, www.nursa.org. ALL, Acute lymphoblastic leukemia; AML, acute myelogenous leukemia; CHO, carbohydrate. ble 1, proteomics strand, project 1) has set out to design and validate a suite of ES cell lines that can be used for tissue-selective overexpression of a targeted subset of commonly studied NRs and coregulators, including members of the SRC/p160 family, PPAR␥ coactivator-1␣ (PGC1␣), and RIP140. In addition to these dedicated resource-generating components, all reagents, protocols, and cell and animal experimental models produced by NURSA-funded research projects are available to the community. Bioinformatics and Web The NURSA bench research projects and resources have generated new information in significant volumes, and the challenge of integrating this with existing information and communicating it effectively to the community has fallen to the McKenna Bioinformatics resource (Table 1, research resource 3). We first established a robust information technology framework within which the various applications which together comprise the NURSA web site (www.nursa.org) could operate. A relational schema was drawn up which encompassed the essential features of NRs, coregulators and ligands as they related to the mission of the consortium (28). We next designed the Molecule Page, A central tenet of the consortium has been to actively engage individuals and organizations from outside the consortium to create sustainable, mutually beneficial relationships that fully exploit the research and intellectual framework established by the Atlas for the benefit of all members of the research community. Community datasets Although NURSA has generated substantial datasets under its own steam, there exist many valuable research resources in the outside community that until now have lacked the backing of a robust, enterprise-grade information technology infrastructure such as NURSA possesses. Our second phase has seen a redoubling of our efforts to identify such datasets and collaborate with the owners to work them into forms suitable for incorporation into the NURSA database. Key examples of community outreach include ongoing collaborations with academic (Tom Scanlan, Oregon Health Sciences University) and industry (GlaxoSmithKline, Wyeth, and Eli Lilly) partners to disseminate information on ligands; working with investigators to develop existing databases for future inclusion in the NURSA database, such as Krish Chatterjee (University of Cambridge) for human PPAR␥ mutations; and forging cross-cutting alliances with complementary web-based resources such as the Phosphosite protein phosphorylation database (http://www.phosphosite.org), the fastDB splicing database (www.fast-db.com), and the MousePAT mouse brain gene expression resource (http://www-mci.u-strasbg.fr/mousepat/). Relationships with publishers A critical component in the future success of NURSA as a community resource is its ability to form alliances with existing scientific publishing organizations, principal among these being established peer-reviewed journals. For example, Molecular En- Mol Endocrinol, June 2009, 23(6):740 –746 docrinology has positioned itself as the preeminent molecular journal in the endocrine field. NURSA annotates Molecular Endocrinology articles accepted for publication, and a collaboration with Highwire Press results in these annotations being displayed on the article web page, such that users have one-click access to a host of contextual information on NRs, coregulators, and ligands relevant to that article. Reciprocally, the NURSA Molecule Pages itemize Molecular Endocrinology papers relevant to each individual curated molecule on the site. In this manner, a synergy is established such that the two formerly distinct web resources become an informational continuum connected by these curated links. Another example of NURSA complementing the traditional roles of established journals is afforded by our ability to take static curated resources from journals, add them to our database, and create functionality that will allow members of the community to update these resources. To illustrate this, resources for animal models of NR (30) and coregulator (8) disruption, originally published in Endocrine Reviews, have been molded into unique living modules of the NURSA Molecule Pages (see Diseases and Phenotypes section of NURSA Molecule Pages). mend.endojournals.org 745 avenues for exploring the roles of NRs and coregulators in health and disease, will prove of lasting value to the broader community of investigators. As a community-based resource, NURSA can significantly enhance the ability of an investigator to initiate, validate, and complete their own lines of inquiry; glean relevant information about NR and coregulator signaling networks; and better inform and catalyze progress in their own research. Acknowledgments We thank all previous members of NURSA including Donald McDonnell, Orla Conneely, Barry Marc Forman, Steve Kliewer, Eric Xu, Adam Antebi, Andrew Dillin, and Yi-Fen Lee and all members of the NR and coregulator signaling community who have contributed to the development of NURSA. Address all correspondence and requests for reprints to: Bert W. O’Malley, M.D., Department of Molecular and Cellular Biology, Baylor College of Medicine, One Baylor Plaza, Houston, Texas 77030. E-mail: berto@bcm.edu; or Ronald M. Evans, Ph.D., Gene Expression Laboratory, The Salk Institute, 10010 North Torrey Pines Road, La Jolla, California 92037. E-mail: evans@salk.edu. Disclosure Summary: The authors have nothing to disclose. Future Directions: Translation The primary mandate facing the NURSA Consortium at this point in its existence is one with which many investigators across the country are increasingly confronted: translation. The overriding goal of the National Institutes of Health is to translate advances in basic science into applications in the clinic. There are many steps between basic science and its application in the clinic, but NURSA has tried to use discovery-based and hypothesis-driven approaches wherever possible to begin moving in this direction. Thus, elucidation of roles for NRs in liver regeneration (31), development of the immune barrier in the small intestine (32), ES cell function (33), and biliary cholestasis (34) all contribute to our understanding of NR and coregulator action in vivo. Furthermore, the discovery of key roles played by members of the orphan receptor subfamily in the development of human cancers, as elucidated in one of our previous pilot projects (35) opens new windows for better understanding of the pathophysiology of disease and potential starting points for study of possible therapeutic intervention. Finally, a study driven by a metabolomics approach, profiling the serum levels of key metabolites in wild-type and knockout animals, has helped identify the SRC/p160 family member SRC-2/Ncoa2 as an eminence grise of carbohydrate metabolism, whose deletion results in a phenotype resembling Von Gierke’s disease of glycogen storage (36). Summary The legacy of NURSA will, we hope, be marked by the creation of this comprehensive Atlas of NRs, coregulators, and ligands. It is our hope that the evolution of the Atlas into an environment for the generation of new hypotheses, generating in turn new References 1. 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