Diethylstilbestrol regulates trophoblast stem cell differentiation as a ligand of orphan nuclear receptor ERRβ

Abstract

The orphan nuclear receptor ERR␤ is expressed in undifferentiated trophoblast stem cell lines and extraembryonic ectoderm, and genetic ablation of ERR␤ results in abnormal trophoblast proliferation and precocious differentiation toward the giant cell lineage. Here, we show that the synthetic estrogen diethylstilbestrol (DES) promotes coactivator release from ERR␤ and inhibits its transcriptional activity. Strikingly, treatment of trophoblast stem cells with DES led to their differentiation toward the polyploid giant cell lineage. In addition, DEStreated pregnant mice exhibited abnormal early placenta development associated with an overabundance of trophoblast giant cells and an absence of diploid trophoblast. These results define a novel pathway for DES action and provide evidence for steroidlike control of trophoblast development. Received December 12, 2000; revised version accepted February 7, 2001. Nuclear receptors constitute a large family of transcription factors that mediate responses to small lipophilic hormones and play essential roles in embryonic development and maintenance of homeostasis in adult animals (Mangelsdorf et al. 1995). Orphan nuclear receptors are members of the nuclear receptor family that lack identified ligands (Giguère 1999). Recent studies have identified several natural and synthetic ligands for a number of orphan nuclear receptors, which led to the discovery of new hormone-response systems implicated in the control of cell fate, organogenesis, and basic metabolic functions Kliewer et al. 1999). While the estrogen-receptor-related receptor (ERR) ␣ (NR3B1) and ERR␤ (NR3B2) were the first orphan nuclear receptors identified more than a decade ago (Giguère et al. 1988), the identification of natural and synthetic ERR ligands has remained elusive. Despite being closely related to the estrogen receptors, ERRs are not activated by natural estrogens (Giguère et al. 1988). In contrast, members of the ERR family, now known to contain a third member referred to as ERR␥ (NR3B3) (Eudy et al. 1998), display various levels of constitutive activity and can interact with coactivators in the absence of ligand Vanacker et al. 1999;Xie et al. 1999).Molecular and genetic studies have shown that ERR␤ plays an important role in early placentation. ERR␤ is expressed in undifferentiated trophoblast stem cell lines (Tanaka et al. 1998) and in a subset of cells in extraembryonic ectoderm destined to make up the chorion (Pettersson et al. 1996;Luo et al. 1997). Genetic ablation of ERR␤ in the mouse results in abnormal chorion formation followed by failure of diploid trophoblast self-renewal and an increase in trophoblast giant cells (Luo et al. 1997). Thus, ERR␤ is essential for normal placental formation, and pharmacological modulation of its activity could influence reproductive function.Diethylstilbestrol (DES) is a potent synthetic estrogen that was used clinically for the prevention of spontaneous abortions from the 1940s to 1971 (Smith 1948). The mechanism of action and factual effectiveness of DES ...

Ube2s regulates Sox2 stability and mouse ES cell maintenance

“…To further clarify whether Esrrb is a downstream effector to confer Ube2s-overexpressed mES cell resistance to differentiation, we treated these cells with diethylstilbestrol (DES) that functions as an estrogen-related receptor antagonist to inhibit Esrrb transcriptional activity. 33 Strikingly, upon adding DES to the RA-containing medium, both red AP staining signals and the expression of selfrenewal genes are reduced in Ube2s-overexpressed mES cells, suggesting that these cells could not resist RA-induced differentiation (Figures 5g and h). These results collectively demonstrate that Ube2s reinforces the undifferentiated state of mES cells through inducing Esrrb.…”

Increased expression of estrogen‐related receptor α (ERRα) is a negative prognostic predictor in human prostate cancer

“…Diethylstilbestrol (DES), an agent widely used for the treatment of advanced PC, represses the molecular activities of ERRa such as reporter gene trans-activation and interaction with co-activator fragments. 42 Thus, the SERMs might inhibit PC by modulating the estrogen-signaling pathway via ERRs besides the ERs. The prognostic value of enhanced ERRa expression as an independent predictor of PC suggests that an ERRa antagonist capable of specifically blocking ERRa activity may prove useful as a therapeutic agent against PC.…”

Elf5-centered transcription factor hub controls trophoblast stem cell self-renewal and differentiation through stoichiometry-sensitive shifts in target gene networks

“…Cells were selected with G418 or Blasticidin S and expanded to 10 15-cm dishes. Cells were washed in PBS, harvested, and used for immunoprecipitation exactly as described previously (van den Berg et al 2010). Precise details are given in the Supplemental Material.…”

“…These lines were used for protein immunoprecipitation (Supplemental Fig. S2C) followed by mass spectrometry (van den Berg et al 2010) to identify Elf5-binding partners in an unbiased manner. Among these, we found factors implicated in trophoblast function, such as the transcriptional regulators Bptf (Goller et al 2008) and Grhl2 (Walentin et al 2015); Sall4, a TF better known for its role in ESC pluripotency (Zhang et al 2006;Yang et al 2008); and components of epigenetic-repressive and nucleosome remodeling complexes (Suz12, Sin3a, and Smca5) ( Fig.…”

“…A number of TFs are known to be involved in either the determination of trophoblast cell fate and/or the ability to derive or maintain TSCs. Among these are the TEA domain family member Tead4 (Yagi et al 2007;Nishioka et al 2008), the caudal-type homeobox factor Cdx2, the Tbox gene Eomes, the SRY-box gene Sox2, and the estrogen-related receptor Esrrb (Russ et al 2000;Tremblay et al 2001;Avilion et al 2003;Strumpf et al 2005;Ralston and Rossant 2008;Adachi et al 2013;Latos et al 2015). Other TFs important for the establishment of a self-renewing TSC population include the AP-2 family member Tfap2c (Auman et al 2002;Werling and Schorle 2002), the Ets family members Ets2 and Elf5 (Yamamoto et al 1998;Donnison et al 2005), and the Gata motif-containing factor Gata3 (Home et al 2009;Ralston et al 2010).…”