Decidualization of human endometrial stromal cells requires steroid receptor coactivator-3

Abstract

Steroid receptor coactivator-3 (SRC-3; also known as NCOA3 or AIB1) is a member of the multifunctional p160/SRC family of coactivators, which also includes SRC-1 and SRC-2. Clinical and cell-based studies as well as investigations on mice have demonstrated pivotal roles for each SRC in numerous physiological and pathophysiological contexts, underscoring their functional pleiotropy. We previously demonstrated the critical involvement of SRC-2 in murine embryo implantation as well as in human endometrial stromal cell (HESC) decidualization, a cellular transformation process required for trophoblast invasion and ultimately placentation. We show here that, like SRC-2, SRC-3 is expressed in the epithelial and stromal cellular compartments of the human endometrium during the proliferative and secretory phase of the menstrual cycle as well as in cultured HESCs. We also found that SRC-3 depletion in cultured HESCs results in a significant attenuation in the induction of a wide-range of established biomarkers of decidualization, despite exposure of these cells to a deciduogenic stimulus and normal progesterone receptor expression. These molecular findings are supported at the cellular level by the inability of HESCs to morphologically transform from a stromal fibroblastoid cell to an epithelioid decidual cell when endogenous SRC-3 levels are markedly reduced. To identify genes, signaling pathways and networks that are controlled by SRC-3 and potentially important for hormone-dependent decidualization, we performed RNA-sequencing on HESCs in which SRC-3 levels were significantly reduced at the time of administering the deciduogenic stimulus. Comparing HESC controls with HESCs deficient in SRC-3, gene enrichment analysis of the differentially expressed gene set revealed an overrepresentation of genes involved in chromatin remodeling, cell proliferation/motility, and programmed cell death. These predictive bioanalytic results were confirmed by the demonstration that SRC-3 is required for the expansion, migratory and invasive activities of the HESC population, cellular properties that are required in vivo in the formation or functioning of the decidua. Collectively, our results support SRC-3 as an important coregulator in HESC decidualization. Since perturbation of normal homeostatic levels of SRC-3 is linked with common gynecological disorders diagnosed in reproductive age women, this endometrial coregulator—along with its new molecular targets described here—may open novel clinical avenues in the diagnosis and/or treatment of a non-receptive endometrium, particularly in patients presenting non-aneuploid early pregnancy loss.

The Long Non-Coding RNA Gene AC027288.3 Plays a Role in Human Endometrial Stromal Fibroblast Decidualization

“…Together, BASP1-AS1 RNA and BASP1 protein form a molecular complex that also includes the ubiquitous bHLH protein TCF12 [73]. The ability of BASP1-AS1 RNA to form a complex with TCF12 is interesting as several bHLH transcription factors play a key role in human endometrial decidualization [27,59,[74][75][76][77][78][79][80][81][82][83]. Targetting LINC01605 expression inhibits autophagy and induces apoptosis in Tenon's capsule fibroblasts [84].…”

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

“…Total RNA was prepared from tissues using the RNeasy Plus Mini kit (Qiagen Inc., Germantown, MD #74134). The NanoDrop 2000 UV/Visual spectrophotometer (ThermoFisher Scientific Inc.) was used for quantification of RNA prior to reverse transcription using the High‐Capacity cDNA Reverse Transcription Kit (ThermoFisher Scientific Inc. #4368814) 30 . Amplified cDNA was diluted to 10 ng/μL before quantitative reverse transcription‐PCR (qRT‐PCR) was performed with the Fast TaqMan 2X Mastermix (Applied Biosystems/Life Technologies, Grand Island, NY #4352042); the TaqMan assays used in this study are listed in Table S1.…”

“…The NanoDrop 2000 UV/Visual spectrophotometer (ThermoFisher Scientific Inc.) was used for quantification of RNA prior to reverse transcription using the High-Capacity cDNA Reverse Transcription Kit (ThermoFisher Scientific Inc. #4368814). 30 Amplified cDNA was diluted to 10 ng/μL before quantitative reverse transcription-PCR (qRT-PCR) was performed with the Fast TaqMan 2X Mastermix (Applied Biosystems/Life Technologies, Grand Island, NY #4352042); the TaqMan assays used in this study are listed in Table S1. All qRT-PCR experiments were performed using the 7500 Fast Real-Time PCR system (Applied Biosystems/ Life Technologies); delta-delta cycle threshold was used to normalize expression to the 18S reference.…”

The Role of SRC-3 in Prostate Cancer Progression and Implications for Therapeutic Targeting: A Systematic Review

“…Among the SRC family members, SRC-3 has garnered increasing attention for its diverse roles in various cancers, including breast, ovarian, and prostate cancer. While initially recognized for its involvement in hormone-dependent breast cancer, SRC-3's significance in prostate cancer is becoming increasingly evident [9]. Prostate cancer's progression is intricately linked to the androgen receptor (AR) signaling pathway.…”