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

Abstract

Progesterone, via the progesterone receptor (PGR), is essential for endometrial stromal cell decidualization, a cellular transformation event in which stromal fibroblasts differentiate into decidual cells. Uterine decidualization supports embryo implantation and placentation as well as subsequent events, which together ensure a successful pregnancy. Accordingly, impaired decidualization results not only in implantation failure or early fetal miscarriage, but also may lead to potential adverse outcomes in all three pregnancy trimesters. Transcriptional reprogramming on a genome-wide scale underlies progesterone dependent decidualization of the human endometrial stromal cell (hESC). However, identification of the functionally essential signals encoded by these global transcriptional changes remains incomplete. Importantly, this knowledge-gap undercuts future efforts to improve diagnosis and treatment of implantation failure based on a dysfunctional endometrium. By integrating genome-wide datasets derived from decidualization of hESCs in culture, we reveal that the promyelocytic leukemia zinc finger (PLZF) transcription factor is rapidly induced by progesterone and that this induction is indispensable for progesterone-dependent decidualization. Chromatin immunoprecipitation followed by next generation sequencing (ChIP-Seq) identified at least ten progesterone response elements within the PLZF gene, indicating that PLZF may act as a direct target of PGR signaling. The spatiotemporal expression profile for PLZF in both the human and mouse endometrium offers further support for stromal PLZF as a mediator of the progesterone decidual signal. To identify functional targets of PLZF, integration of PLZF ChIP-Seq and RNA Pol II RNA-Seq datasets revealed that the early growth response 1 (EGR1) transcription factor is a PLZF target for which its level of expression must be reduced to enable progesterone dependent hESC decidualization. Apart from furnishing essential insights into the molecular mechanisms by which progesterone drives hESC decidualization, our findings provide a new conceptual framework that could lead to new avenues for diagnosis and/or treatment of adverse reproductive outcomes associated with a dysfunctional uterus.

Using CRISPR/Cas9 engineering to generate a mouse with a conditional knockout allele for the promyelocytic leukemia zinc finger transcription factor

“…Genotyping with PCR primers: P1, P2, and P3 (Table and Supporting Information Figure S2), demonstrated absence of exon 2 in the Plzf d/d mouse (Figure a,b). Because we have previously demonstrated that Plzf protein is significantly induced in uterine tissue of ovariectomized wild type mice following injection of progesterone hormone (Kommagani et al, ), we injected ovariectomized Plzf d/d females and corresponding monogenic controls with either hormone vehicle or progesterone hormone. Western analysis clearly showed that while uterine Plzf protein is not expressed in the vehicle‐treated ovariectomized control and Plzf d/d group, Plzf protein expression is only induced in the control uterus 6 hr following progesterone administration (Figure c).…”

“…Lanes 1 and 2 represent protein isolated from the uterus of untreated and progesterone‐treated ovariectomized wild type mice, respectively. Note the significant induction of Plzf protein (75 kDa) expression (after 6‐hr) in response to progesterone administration (Kommagani et al, ). Lanes 3 and 4 represent uterine protein isolates from untreated and progesterone‐treated Plzf d/d mice, respectively.…”

“…We predict that the Plzf f/f mouse will serve as an invaluable tool with which to further elucidate Plzf function in a tissue and cell‐type specific manner. For example, we recently demonstrated that PLZF is essential for human endometrial stromal cell decidualization (Kommagani et al, ; Szwarc et al, ), a cellular process required for embryo implantation. Crossing the Plzf f/f mouse with our previously reported progesterone receptor promoter‐driven cre ( Pgr cre ) knockin mouse (Soyal et al, ) will now allow us to functionally validate Plzf action specifically in the mouse endometrium, without interference of phenotypes contributed by other target tissues due to this transcription factor's loss‐of‐function.…”

“…Positioned within the C‐terminal region of PLZF, the nine C2H2 zinc finger motifs facilitate direct sequence‐specific DNA binding to target genes. Cell and signaling context dependent, PLZF can serve as a transcriptional activator or repressor (Fahnenstich et al, ; Gaber, Butler, & Novitch, ; Ikeda et al, ; Kommagani et al, ; Labbaye et al, ; Liu et al, ; Mao et al, ; Singh et al, ; Suliman et al, ; Szwarc et al, ; Wang et al, ). Given its pleiotropic effects, PLZF drives a broad spectrum of developmental processes and physiological responses, including limb skeletal patterning, innate immune cell development, hematopoiesis, and spermatogenesis; reviewed in Suliman et al ().…”

New models of lipopolysaccharide‐induced implantation loss reveal insights into the inflammatory response

“…Transcript levels of insulin-like growth factor binding protein-1 (IGFBP1) and prolactin (PRL) were used as markers for the decidual response in hESCs 20. hESCs were grown in DMEM media supplemented with heat-inactivated 10% FBS.…”