Reporter Cell Lines for the Characterization of the Interactions between Human Nuclear Receptors and Endocrine Disruptors Marina Grimaldi, Abdelhay Boulahtouf, Vanessa Delfosse, Erwan Thouennon, William Bourguet, Patrick Balaguer To cite this version: Marina Grimaldi, Abdelhay Boulahtouf, Vanessa Delfosse, Erwan Thouennon, William Bourguet, et al.. Reporter Cell Lines for the Characterization of the Interactions between Human Nu- clear Receptors and Endocrine Disruptors. Frontiers in Endocrinology, Frontiers, 2015, 6, pp.62. 10.3389/fendo.2015.00062. hal-02165218 HAL Id: hal-02165218 https://hal.umontpellier.fr/hal-02165218 Submitted on 25 Jun 2019 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. 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Distributed under a Creative Commons Attribution - NonCommercial| 4.0 International License REVIEW published: 11 May 2015 doi: 10.3389/fendo.2015.00062 Reporter cell lines for the characterization of the interactions between human nuclear receptors and endocrine disruptors Marina Grimaldi 1,2,3,4, Abdelhay Boulahtouf 1,2,3,4, Vanessa Delfosse 3,5,6, Erwan Thouennon 1,2,3,4, William Bourguet 3,5,6 and Patrick Balaguer 1,2,3,4* 1 IRCM, Institut de Recherche en Cancérologie de Montpellier, Montpellier, France, 2 U1194, INSERM, Montpellier, France, 3 Université Montpellier, Montpellier, France, 4 ICM, Institut Régional du Cancer de Montpellier, Montpellier, France, 5 U1054, INSERM, Montpellier, France, 6 CNRS UMR5048, Centre de Biochimie Structurale, Montpellier, France Endocrine-disrupting chemicals (EDCs) are exogenous substances interfering with hor- mone biosynthesis, metabolism, or action, and consequently causing disturbances in the endocrine system. Various pathways are activated by EDCs, including interactions with nuclear receptors (NRs), which are primary targets of numerous environmental Edited by: contaminants. The main NRs targeted by environmental contaminants are the estrogen David H. Volle, α β INSERM, France (ER , ) and the androgen (AR) receptors. ERs and AR have pleiotropic regulatory roles Reviewed by: in a diverse range of tissues, notably in the mammary gland, the uterus, and the prostate. Isaias Glezer, Thus, dysfunctional ERs and AR signaling due to inappropriate exposure to environmental Universidade Federal de São Paulo, Brazil pollutants may lead to hormonal cancers and infertility. The pregnane X receptor (PXR) is Jean-Marc A. Lobaccaro, also recognized by many environmental molecules. PXR has a protective role of the body Clermont Université, France through its ability to regulate proteins involved in the metabolism, the conjugation, and the *Correspondence: transport of many exogenous and endogenous compounds. However, the permanent Patrick Balaguer, U1194, IRCM, INSERM, ICM, Parc activation of this receptor by xenobiotics may lead to premature drug metabolism, the Euromédecine, 208 rue des formation, and accumulation of toxic metabolites and defects in hormones homeostasis. Apothicaires, Montpellier 34090, France The activity of other NRs can also be affected by environmental molecules. Compounds [email protected] capable of inhibiting or activating the estrogen related (ERRγ), the thyroid hormone (TRα, β), the retinoid X receptors (RXRα, β, γ), and peroxisome proliferator-activated (PPAR α, Specialty section: γ) receptors have been identified and are highly suspected to promote developmental, This article was submitted to Cellular Endocrinology, a section of the journal reproductive, neurological, or metabolic diseases in humans and wildlife. In this review, Frontiers in Endocrinology we provide an overview of reporter cell lines established to characterize the human NR Received: 02 February 2015 activities of a large panel of EDCs including natural as well as industrial compounds such Accepted: 09 April 2015 as pesticides, plasticizers, surfactants, flame retardants, and cosmetics. Published: 11 May 2015 Citation: Keywords: nuclear receptors, environmental-disrupting compounds, reporter cell lines Grimaldi M, Boulahtouf A, Delfosse V, Thouennon E, Bourguet W and Balaguer P (2015) Reporter cell lines for the characterization of the Introduction interactions between human nuclear receptors and endocrine disruptors. Human nuclear receptors (NRs) are a family of 48 transcription factors, many of which have Front. Endocrinol. 6:62. been shown to be activated by endogenous ligands. NRs regulate cognate gene networks doi: 10.3389/fendo.2015.00062 involved in key physiological functions such as cell growth and differentiation, development, Frontiers in Endocrinology | www.frontiersin.org 1 May 2015 | Volume 6 | Article 62 Grimaldi et al. Nuclear receptor reporter cell lines A A/B C D E F Nt (AF-1) DBD Hinge LBD (AF-2) Ct B NR dimer SIGNAL HSP protein NR/hormone Ligand complex Target gene Type I NR RE HSP/NR + Transcriptional machinery mRNA complex RXR Target gene RXR-NR heterodimer Type 2 NR RE FIGURE 1 | General nuclear receptor structure and function. (A) Structural complexes with chaperone proteins in the cytoplasm (AR) or in the nucleus (ERs) organization of NRs. NRs comprise six domains, including a N-terminal whereas type II NRs (RXR heterodimers) are bound to their target genes with activation function domain, a central DNA binding domain, and a C-terminal corepressors. Ligand binding results in the dissociation of chaperone proteins ligand-binding domain carrying a ligand-dependant transcriptional function. (B) and binding and activation to target genes for type I NRs. Ligand binding results Schematic model of NR function. Before ligand binding, type I NRs form inactive in corepressors release and coactivator recruitment for type II NRs. homeostasis, or metabolism (1, 2). As a consequence, inappropri- TABLE 1 | Nuclear receptor characterization. ate exposure to environmental pollutants, which have the ability NR Cellular localization Active form to substitute for natural ligands, can cause proliferative, repro- in absence of ligand ductive, and metabolic disorders, including hormone-dependent cancers, infertility, diabetes, or obesity. ERα (NR3A1) Nuclear Homodimer NRs are transcriptional regulators comprising several domains, ERβ (NR3A2) Nuclear Homodimer including a N-terminal activation function domain (AF-1), a AR (NR3C4) Cytoplasmic Homodimer ERRγ (NR3B3) Nuclear Monomer central DNA-binding domain (DBD), and a C-terminal ligand- PPARα (NR1C1) Nuclear RXR heterodimer binding domain (LBD) carrying a ligand-dependent transcrip- PPARγ (NR1C3) Nuclear RXR heterodimer tional activation function (AF-2) (2)(Figure 1A). When unas- TRα (NR1A1) Nuclear RXR heterodimer β sociated with their ligand, type I NRs form inactive complexes TR (NR1A2) Nuclear RXR heterodimer PXR (NR1I2) Nuclear RXR heterodimer with chaperone proteins in the cytoplasm, whereas type II NRs are RXRα (NR2B1) Nuclear Heterodimer or homodimer located in the nucleus and bind to the DNA response elements of RXRβ (NR2B2) Nuclear Heterodimer or homodimer their target genes along with corepressors (Table 1; Figure 1B). RXRγ (NR2B3) Nuclear Heterodimer or homodimer Ligand binding triggers major conformational changes in the receptor LBD that lead to the dissociation of chaperones and core- pressors, nuclear translocation and DNA binding of type I NRs, association with coactivators and enables corepressors recruit- and recruitment of coactivators, thus initiating gene transcription. ment (3–5). The LBD also contributes to the modulation of the In presence of agonists in the ligand-binding pocket, corepressors N-terminal AF-1 through interdomain crosstalk, which enable dissociate and the recruitment of transcriptional coactivators is AF-1 and AF-2 domains to recruit coactivators individually or in favored (3–5). Reciprocally, interaction with antagonists avoids a synergistic manner (6–8). Frontiers in Endocrinology | www.frontiersin.org 2 May 2015 | Volume 6 | Article 62 Grimaldi et al. Nuclear receptor reporter cell lines Endocrine-disrupting chemicals (EDCs) are exogenous sub- liver, and heart, whereas ERβ is primarily expressed in the ovary, stances that interfere with the function of hormonal systems prostate, gastrointestinal tract, lung, bladder, and hematopoietic and produce a range of developmental, reproductive, neurolog- and central nervous systems (16). However, ERα and ERβ are ical, immune, or metabolic diseases in humans and wildlife (9, also coexpressed in numerous tissues such as the mammary gland, 10). Many EDCs are man-made chemicals produced by indus- adrenal, thyroid, bones, and some regions of the brain. try and released into the environment as, for example, bisphe- Interestingly, when ERs are coexpressed, ERβ exhibits an nols, phthalates, pesticides, organotins, flame retardants, poly- inhibitory action on ERα-mediated gene expression (17, 18) so chlorinated biphenyls, dioxins, or alkylphenols. EDCs can also that ERβ has been shown to antagonize several ERα-mediated be naturally produced by plants or fungus, like the genistein and effects including fat reduction and cellular proliferation in breast, daidzein phytoestrogens,