Early growth response 1 transcription factor is essential for the pathogenic properties of human endometriotic epithelial cells

Abstract

Although a benign gynecological disorder, endometriosis displays some pathogenic features of malignancy, such as cell proliferation, migration, invasion, and adaptation to hypoxia. Current treatments of endometriosis include pharmacotherapy and/or surgery, which are of limited efficacy and often associated with adverse side-effects. Therefore, to develop more effective therapies to treat this disease, a broader understanding of the underlying molecular mechanisms that underpin endometriosis needs to be attained. Using immortalized human endometriotic epithelial and stromal cell lines, we demonstrate that the early growth response 1 (EGR1) transcription factor is essential for cell proliferation, migration and invasion, which represent some of the pathogenic properties of endometriotic cells. Genome-wide transcriptomics identified an EGR1-dependent transcriptome in human endometriotic epithelial cells that potentially encodes a diverse spectrum of proteins that are known to be involved in tissue pathologies. To underscore the utility of this transcriptomic dataset, we demonstrate that carbonic anhydrase IX (CAIX), a homeostatic regulator of intracellular pH, is not only a molecular target of EGR1 but is important for maintaining many of the cellular properties of human endometriotic epithelial cells that are also ascribed to EGR1. Considering therapeutic intervention strategies are actively being developed for EGR1 and CAIX in the treatment of other pathologies, we believe EGR1 and its transcriptome (which includes CAIX) will offer not only a new conceptual framework to advance our understanding of endometriosis but will furnish new molecular vulnerabilities to be leveraged as potential therapeutic options in the future treatment of endometriosis.

A CRISPR/Cas9‐engineered mouse carrying a conditional knockout allele for the early growth response‐1 transcription factor

“…Collectively, our genetic, protein, and phenotypic analysis in this short technical report demonstrates that the Egr1 f/f allele operates as designed and therefore will be of significant utility for researchers interested in resolving EGR1 responses at a tissue and/or cell‐type specific level. For example, we have recently demonstrated that EGR1 is critical for both normal and abnormal uterine function (Maurya et al, 2022; Park et al, 2021; Szwarc et al, 2019). As EGR1 is expressed in endometrial epithelial and stromal cells (Maurya et al, 2022; Park et al, 2021; Szwarc et al, 2019), selective ablation of the EGR1 in the epithelial or stromal‐specific cell type will provide an unprecedented opportunity to investigate EGR1's role in endometrial biology and pathobiology at a cell‐type specific level.…”

“…For example, we have recently demonstrated that EGR1 is critical for both normal and abnormal uterine function (Maurya et al, 2022; Park et al, 2021; Szwarc et al, 2019). As EGR1 is expressed in endometrial epithelial and stromal cells (Maurya et al, 2022; Park et al, 2021; Szwarc et al, 2019), selective ablation of the EGR1 in the epithelial or stromal‐specific cell type will provide an unprecedented opportunity to investigate EGR1's role in endometrial biology and pathobiology at a cell‐type specific level. In closing, it should be noted that the use of an Egr1 f/f mouse was recently reported by a research group in China (Lei et al, 2022; Wu et al, 2018, 2019); however, details on how this mouse was generated or whether the mouse will be made available to the scientific community has not been published.…”

“…Also known as NGFI‐A, ZIF 268, KROX 24, or TIS8 (Thiel & Cibelli, 2002), EGR1 is a modular domain nuclear protein and a founding member of the evolutionary conserved EGR family of Cys 2 ‐His 2 zinc finger (ZF) transcription factors, which includes EGR2, EGR3, and EGR4 (Gashler & Sukhatme, 1995). Through binding cognate GC‐rich (5′‐GCG(T/G)GGGCG‐3′) DNA motifs via its three ZF DNA binding domains (Swirnoff & Milbrandt, 1995), EGR1 directly activates or represses transcription of a diverse range of target genes, which, in turn, control a myriad of cellular processes, including (but not limited to) cell division, migration, adhesion, matrix deposition, and thrombosis (Adamson & Mercola, 2002; Bhattacharyya et al, 2011; Blaschke, Bruemmer, & Law, 2004; DeLigio & Zorio, 2009; Du, Wu, Ji, Liang, & Li, 2020; Maurya et al, 2022).…”

Steroid receptor coactivator‐2 drives epithelial reprogramming that enables murine embryo implantation

“…Genome scale RNA‐sequencing (RNA‐seq) and analysis were performed as previously described 31,32 . Briefly, total RNA purity and physical integrity were determined using the NanoDrop spectrophotometer (ThermoFisher Scientific Inc.) and the 2100 Bioanalyzer with RNA chips (Agilent Technologies Inc., Santa Clara, CA), respectively.…”

“…Genome scale RNA-sequencing (RNA-seq) and analysis were performed as previously described. 31,32 Briefly, total RNA purity and physical integrity were determined using the NanoDrop spectrophotometer (ThermoFisher Scientific Inc.) and the 2100 Bioanalyzer with RNA chips (Agilent Technologies Inc., Santa Clara, CA), respectively. Total RNA preparations that scored a RNA integrity number (RIN) of 7 or greater were used in these experiments; RNA samples from three mice were used for each genotype.…”