Yilmaz Bahar D, Sison Christia A M, Yildiz Sule, Miyazaki Kaoru, Coon V John, Yin Ping, Bulun Serdar E
Division of Reproductive Science and Medicine, Department of Obstetrics and Gynecology, Northwestern University Feinberg School of Medicine, Chicago, Illinois.
Division of Reproductive Science and Medicine, Department of Obstetrics and Gynecology, Northwestern University Feinberg School of Medicine, Chicago, Illinois.
F S Sci. 2020 Aug;1(1):59-66. doi: 10.1016/j.xfss.2020.06.002.
To investigate the gene targets of estradiol (E2)-estrogen receptor-α (ESR1) in human endometrial stromal cells.
Basic science.
University research center.
PATIENT(S): Premenopausal women with or without endometriosis.
INTERVENTION(S): Primary cultures of human endometrial stromal cells from healthy endometrium, with or without small-interfering RNA (siRNA) knockdown of ESR1 expression, were treated with E2 or vehicle control.
MAIN OUTCOME MEASURE(S): Genome-wide RNA expression by RNA sequencing was compared in endometrial stromal cells with or without siRNA knockdown of ESR1 in the presence or absence of E2. Genome-wide recruitment of ESR1 to chromatin was assessed by chromatin immunoprecipitation sequencing. Gene expression by real-time qualitative polymerase chain reaction of a potential E2-ESR1 target gene was determined in endometrial stromal cells and endometriotic stromal cells.
RESULT(S): We identified several important pathways that are dependent on E2-ESR1 signaling in endometrial stromal cells, including progesterone signaling, cell-matrix adhesion, and cytoskeleton rearrangement, as well as paracrine signaling by members of the fibroblast growth factor family. We detected a total of 709 ESR1 target sites on chromatin. By integrating data on genome-wide transcriptomic changes and E2-ESR1 binding sites, we identified inositol polyphosphate phosphatase type II (INPP4B) as a candidate E2-mediated suppressor of proliferation in healthy endometrial cells. INPP4B was downregulated in endometriosis-derived stromal cells.
CONCLUSION(S): E2-ESR1 activates genes involved in human endometrial stromal cell cycle regulation, progesterone response, and production of stromal growth factors. Understanding the direct role of estrogen on the endometrial stroma and identifying downstream targets of E2-ESR1 can inform the development of targeted therapies for endometriosis and diminished endometrial receptivity.
研究雌二醇（E2）-雌激素受体-α（ESR1）在人子宫内膜基质细胞中的基因靶点。
基础科学研究。
大学研究中心。
有或无子宫内膜异位症的绝经前女性。
从健康子宫内膜获取人子宫内膜基质细胞进行原代培养，在有或无小干扰RNA（siRNA）敲低ESR1表达的情况下，用E2或载体对照进行处理。
在有或无E2的情况下，比较有或无siRNA敲低ESR1的子宫内膜基质细胞中通过RNA测序得到的全基因组RNA表达。通过染色质免疫沉淀测序评估ESR1在全基因组范围内与染色质的结合情况。在子宫内膜基质细胞和子宫内膜异位症基质细胞中，通过实时定量聚合酶链反应测定潜在的E2-ESR1靶基因的表达。
我们确定了子宫内膜基质细胞中依赖E2-ESR1信号传导的几个重要途径，包括孕酮信号传导、细胞-基质黏附、细胞骨架重排，以及成纤维细胞生长因子家族成员的旁分泌信号传导。我们在染色质上共检测到709个ESR1靶位点。通过整合全基因组转录组变化数据和E2-ESR1结合位点数据，我们确定肌醇多磷酸磷酸酶II型（INPP4B）是健康子宫内膜细胞中E2介导的增殖抑制因子的候选基因。INPP4B在子宫内膜异位症来源的基质细胞中表达下调。
E2-ESR1激活参与人子宫内膜基质细胞周期调节、孕酮反应和基质生长因子产生的基因。了解雌激素对子宫内膜基质的直接作用并确定E2-ESR1的下游靶点可为子宫内膜异位症和子宫内膜容受性降低的靶向治疗开发提供依据。
