Fabrice G. Petit

COUP-TFII Mediates Progesterone Regulation of Uterine Implantation by Controlling ER Activity

Progesterone and estrogen are critical regulators of uterine receptivity. To facilitate uterine remodeling for embryo attachment, estrogen activity in the uterine epithelia is attenuated by progesterone; however, the molecular mechanism by which this occurs is poorly defined. COUP-TFII (chicken ovalbumin upstream promoter transcription factor II; also known as NR2F2), a member of the nuclear receptor superfamily, is highly expressed in the uterine stroma and its expression is regulated by the progesterone–Indian hedgehog–Patched signaling axis that emanates from the epithelium. To further assess COUP-TFII uterine function, a conditional COUP-TFII knockout mouse was generated. This mutant mouse is infertile due to implantation failure, in which both embryo attachment and uterine decidualization are impaired. Using this animal model, we have identified a novel genetic pathway in which BMP2 lies downstream of COUP-TFII. Epithelial progesterone-induced Indian hedgehog regulates stromal COUP-TFII, which in turn controls BMP2 to allow decidualization to manifest in vivo. Interestingly, enhanced epithelial estrogen activity, which impedes maturation of the receptive uterus, was clearly observed in the absence of stromal-derived COUP-TFII. This finding is consistent with the notion that progesterone exerts its control of implantation through uterine epithelial-stromal cross-talk and reveals that stromal-derived COUP-TFII is an essential mediator of this complex cross-communication pathway. This finding also provides a new signaling paradigm for steroid hormone regulation in female reproductive biology, with attendant implications for furthering our understanding of the molecular mechanisms that underlie dysregulation of hormonal signaling in such human reproductive disorders as endometriosis and endometrial cancer.

Deletion of the orphan nuclear receptor COUP-TFII in uterus leads to placental deficiency

COUP-TFII (NR2F2), chicken ovalbumin upstream promoter–transcription factor II, is an orphan nuclear receptor of the steroid/thyroid hormone receptor superfamily. The Coup-tfII-null mutant mice die during the early embryonic development because of angiogenesis and heart defects. To analyze the physiological function of COUP-TFII during organogenesis, we used the cre/loxP system to conditionally inactivate COUP-TFII in the ovary and uterus. Homozygous adult female mutants with specific inactivation of the Coup-tfII gene in uterine stromal and smooth muscle cells have severely impaired placental formation, leading to miscarriage at days 10–12 of pregnancy. Deletion of the Coup-tfII gene resulted in an increase in trophoblast giant cell differentiation, a reduction of the spongiotrophoblast layer, and an absence of labyrinth formation causing an improper vascularization of the placenta. This study describes an important maternal role of COUP-TFII in regulating the placentation. The endometrial COUP-TFII might modulate the signaling between the uterus and the extraembryonic tissue for the proper formation of the placenta.

The regulation of COUP-TFII gene expression by Ets-1 is enhanced by the steroid receptor co-activators.

Recent phenotypic analysis of orphan nuclear receptor chicken ovalbumin upstream promoter-transcription factor II (COUP-TFII) [NR2F2] knockout mice shows that COUP-TFII is involved in the angiogenic process in the developing embryos. Since Ets-1 expression is also correlated with angiogenesis, and both Ets-1 and COUP-TFII mRNA are present in mesenchymal cells, we have sought to determine whether Ets-1 is a potential regulator of COUP-TFII gene expression. For this purpose, we performed transient transfection experiments using a luciferase reporter construct containing the mouse COUP-TFII promoter. We found that the COUP-TFII promoter activity is indeed regulated by Ets-1. We have identified two identical inverted potential ETS-binding sites located 47 nucleotides downstream of the start site. Mutation of both sites reduced the ability of Ets-1 to enhance the COUP-TFII promoter activity. Furthermore, other members of the ETS family such as Ets-2 or ETV1 are also potent regulators of the COUP-TFII promoter. Finally, the induction of the COUP-TFII gene is strongly enhanced by the expression of steroid receptor co-activator factors through a direct interaction with Ets-1. These results indicate that COUP-TFII is a potential downstream target of Ets-1 and it may partially mediate the Ets-1 function in angiogenesis.

Partial Müllerian Duct Retention in Smad4 Conditional Mutant Male Mice.

Müllerian duct regression is a complex process which involves the AMH signalling pathway. We have previously demonstrated that besides AMH and its specific type II receptor (AMHRII), BMPR-IA and Smad5 are two essential factors implicated in this mechanism. Mothers against decapentaplegic homolog 4 (Smad4) is a transcription factor and the common Smad (co-Smad) involved in transforming growth factor beta (TGF-β) signalling pathway superfamily. Since Smad4 null mutants die early during gastrulation, we have inactivated Smad4 in the Müllerian duct mesenchyme. Specific inactivation of Smad4 in the urogenital ridge leads to the partial persistence of the Müllerian duct in adult male mice. Careful examination of the urogenital tract reveals that the Müllerian duct retention is randomly distributed either on one side or both sides. Histological analysis shows a uterus-like structure, which is confirmed by the expression of estrogen receptor α. As previously described in a β-catenin conditional mutant mouse model, β-catenin contributes to Müllerian duct regression. In our mutant male embryos, it appears that β-catenin expression is locally reduced along the urogenital ridge as compared to control mice. Moreover, the expression pattern is similar to those observed in control female mice. This study shows that reduced Smad4 expression disrupts the Wnt/β-catenin signalling leading to the partial persistence of Müllerian duct.

CHAPTER 279 – Orphan Receptor COUP-TFII and Vascular Development

Nuclear receptors belong to a large family of transcription factors, which can modulate gene expression. Since the discovery of the steroid/thyroid hormone receptor superfamily, an increasing number of transcription factors with similar structural motif have been added. A majority of these newly added members have unknown ligands, thus they are classified as orphan nuclear receptors. Like the other members of the steroid/thyroid hormone receptor superfamily, the orphan receptors are involved in development, differentiation, and homeostasis processes. The absence of ligand makes it more difficult to dissect the functional role of orphan receptors. However, the analysis of orphan receptor null mice will provide new insights into the physiological function of these receptors during development, organogenesis, and homeostasis. This chapter attempts to describe the functional role of orphan receptors during vascular development.