A Gata2-Dependent Transcription Network Regulates Uterine Progesterone Responsiveness and Endometrial Function

Abstract

SUMMARY Altered progesterone responsiveness leads to female infertility and cancer, but underlying mechanisms remain unclear. Mice with uterine-specific ablation of Gata2 are infertile showing failures in embryo implantation, endometrial decidualization and uninhibited estrogen signaling. Gata2 deficiency results in reduced progesterone receptor (PGR) expression and attenuated progesterone signaling, as evidenced by genome-wide expression profiling and chromatin immunoprecipitation. GATA2 not only occupies at and promotes expression of Pgr, but also regulates downstream progesterone responsive genes in conjunction with the PGR. Additionally, Gata2 knockout uteri exhibit abnormal luminal epithelia with ectopic TRP63 expressing squamous cells and a cancer related molecular profile in a progesterone independent manner. Lastly, we found a conserved GATA2-PGR regulatory network in both human and mouse, based on gene signature and path analyses using gene expression profiles of human endometrial tissues. In conclusion, uterine Gata2 regulates a key regulatory network of gene expression for progesterone signaling at the early pregnancy stage.

Reconstructing Lineage Hierarchies of Mouse Uterus Epithelial Development Using Single-Cell Analysis

“…Most of the other TFs identified herein are not studied in the development of uterine epithelia. We found that Gata2 (Rubel et al., 2012, Rubel et al., 2016) was uniquely expressed by P14 cells. Immunocytochemistry was performed to confirm the expression of GATA2 at the protein level, with the results showing the highest expression of GATA2 protein in the P14 cells, with particularly strong expression in the cell nuclei (Figure 3B).

Figure 3Molecular Cascades Regulating the Development and Maturation of Uterine Epithelia(A) Differentially expressed transcription factors (TFs) of the respective cell cluster were selected and visualized using a heatmap.(B) Expression of GATA2 protein in uterine epithelia (CD326 + ) during development.

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Hormone dependent uterine epithelial-stromal communication for pregnancy support

“…Recently, new gene editing technology, such as CRISPR/Cas9 system has opened a new era for the generation of transgenic animal models in a more efficient and economical way that can help further identify uterine-specific enhancer or promoter regions of these PGR-associated genes. For example, PGR and GATA2 both occupy at putative enhancers of the Sox17 and Ihh genes [45] that have pivotal uterine functions [54, 91, 92]. While results from in vitro assays suggest a cooperation of PGR and GATA2 in the regulation of enhancers for the Sox17 and Ihh genes, the in vivo function of such enhancers remains to be demonstrated, perhaps by CRISPR technology.…”

“…While results from in vitro assays suggest a cooperation of PGR and GATA2 in the regulation of enhancers for the Sox17 and Ihh genes, the in vivo function of such enhancers remains to be demonstrated, perhaps by CRISPR technology. Furthermore, advanced statistical and bioinformatics tools are available for integrated analysis of genome-wide data from both human subjects and mouse models, which can help to identify genetic networks and regulatory mechanisms that are conserved across species [45, 93–95]. Importantly, mapping out evolutionary conservation and difference on regulatory pathways would help better interpretation on data generated from preclinical investigations.…”

“…Mice with germline ablation of Gata2 display defects in hematopoietic cell development [41], vascular integrity [42], adipocyte differentiation [43], and pituitary function [44]. Mice with uterine ablation of Gata2 ( Pgr Cre/+ Gata2 flox/flox ) are infertile with defects in conceptus implantation and subsequent uterine stromal decidualization [45]. These mice display stratification in uterine epithelia with increased TRP63 expression, as well as, reduction in PGR target genes and Pgr itself.…”

“…Further cistromic analysis with transcriptomic analysis identified a distinct uterine-specific GATA2 genomic occupancy profile that encompasses most PGR target genes, suggesting a cooperative relationship between the two factors in controlling P4-mediated transcription in the mouse uterus. Importantly, this GATA2-PGR regulatory axis is also observed in endometrial specimens of human subjects consisting of normal and recurrent pregnancy loss patients, which provides a clinical indication and evidence of evolutional conservation of this pathway [45]. …”