A Murine Uterine Transcriptome, Responsive to Steroid Receptor Coactivator-2, Reveals Transcription Factor 23 as Essential for Decidualization of Human Endometrial Stromal Cells1

Abstract

Recent data from human and mouse studies strongly support an indispensable role for steroid receptor coactivator-2 (SRC-2)-a member of the p160/SRC family of coregulators-in progesterone-dependent endometrial stromal cell decidualization, an essential cellular transformation process that regulates invasion of the developing embryo into the maternal compartment. To identify the key progesterone-induced transcriptional changes that are dependent on SRC-2 and required for endometrial decidualization, we performed comparative genome-wide transcriptional profiling of endometrial tissue RNA from ovariectomized SRC-2(flox/flox) (SRC-2(f/f) [control]) and PR(cre/+)/SRC-2(flox/flox) (SRC-2(d/d) [SRC-2-depleted]) mice, acutely treated with vehicle or progesterone. Although data mining revealed that only a small subset of the total progesterone-dependent transcriptional changes is dependent on SRC-2 (∼13%), key genes previously reported to mediate progesterone-driven endometrial stromal cell decidualization are present within this subset. Along with providing a more detailed molecular portrait of the decidual transcriptional program governed by SRC-2, the degree of functional diversity of these progesterone mediators underscores the pleiotropic regulatory role of SRC-2 in this tissue. To showcase the utility of this powerful informational resource to uncover novel signaling paradigms, we stratified the total SRC-2-dependent subset of progesterone-induced transcriptional changes in terms of novel gene expression and identified transcription factor 23 (Tcf23), a basic-helix-loop-helix transcription factor, as a new progesterone-induced target gene that requires SRC-2 for full induction. Importantly, using primary human endometrial stromal cells in culture, we demonstrate that TCF23 function is essential for progesterone-dependent decidualization, providing crucial translational support for this transcription factor as a new decidual mediator of progesterone action.

The Promyelocytic Leukemia Zinc Finger Transcription Factor Is Critical for Human Endometrial Stromal Cell Decidualization

“…As further support for this conclusion, immunohistochemical analysis of human endometrial biopsy samples show that PLZF protein is highly expressed in the nucleus of the endometrial stromal cell during the P4-dominant secretory phase of the menstrual cycle but is expressed at low levels in endometrial tissue biopsied during the E2-dominant proliferative phase of the cycle ( Fig 1D–1F ). In the uterus of the ovariectomized mouse, Plzf transcript and protein levels are also significantly and rapidly induced by P4 ( S2 Fig ), validating our previous microarray analysis of this tissue which showed Plzf transcription is induced by short-term P4 exposure [ 9 ]. Transcript and protein levels for Plzf were detected as early as six hours following P4 administration ( S2A and S2B Fig ).…”

“…Comparing undecidualized hESCs with decidualized hESCs in culture, we recently revealed by RNA-seq analysis that PLZF (also known as ZBTB 16 , a Kruppel C2H2-type zinc-finger transcription factor) is listed in the top 20 genes which are significantly induced in hESCs during decidualization [ 8 ] ( S1A Fig ). Additionally, we previously showed by microarray analysis that Plzf is present in the top 100 genes induced in total uterine tissue of the mouse in response to acute P4 administration [ 9 ], suggesting PLZF is a P4-responsive gene and important for decidualization. To address this proposal, an established cell culture-based model for decidualization of primary hESCs was used to determine the transcriptional induction profile of PLZF ( Fig 1A ).…”

“…A portion of the endometrial biopsy was used for immunohistochemistry (see below). The remainder of the biopsy was used to prepare human endometrial stromal cells (hESCs) as previously described [ 9 , 62 ]. Isolated hESCs were cultured in DMEM/F-12 media containing 10% fetal bovine serum (FBS), 100 units/ml penicillin, and 0.1 mg/ml streptomycin (termed hESC media); studies were conducted with early passage primary hESCs (≤ four passages).…”

Deletion of Arid1a in Reproductive Tract Mesenchymal Cells Reduces Fertility in Female Mice1

“…Primary cultures of human endometrial stromal fibroblasts (hESCs) have the ability to undergo in vitro decidualization, and this culture system has been used to demonstrate the importance of several factors in uterine decidualization, such as LIF [ 84 ], transient receptor potential cation channel, subfamily C, member 1 ( TRPC1 ) [ 85 ], transcription factor 23 ( TCF23 ) [ 86 ], uterine activing like kinase 2 (ALK2) [ 78 ], bone morphogenetic protein 2 ( BMP2 ) [ 87 ], and WNT4 [ 88 ]. Previous work has shown that ARID1A is deficient in hESCs from women with endometriosis [ 21 ] and that in vitro decidualization of hESCs from women with endometriosis have a blunted in vitro decidualization response [ 89 ].…”

The p160/Steroid Receptor Coactivator Family: Potent Arbiters of Uterine Physiology and Dysfunction1

“…4) Apart from PFKFB3 induction, does SRC-2 coregulate other progesterone signaling pathways that are important for ESC decidualization? In the case of the mouse at least, recent microarray array studies reveal that SRC-2 is critically important for the full induction of the majority of previously known molecular mediators of progesterone-driven ESC decidualization [110]. Whether these findings can be translated to the human ESC remains an open question.…”