Endometriosis and Retinoids

子宫内膜异位症是盆腔器官上的病理性子宫内膜样组织，其发展和持续存在的部分原因是 细胞凋亡缺陷，并与盆腔疼痛和不孕症有关。我们的总体假设是有缺陷的 类固醇激素和RA信号转导与生殖内分泌细胞的异常重编程有关。我们 提出三个目标。1)确定Rorb缺乏，黄体酮抵抗的结果 对子宫内膜异位症细胞存活和分化的改变负责。我们发现了一种新的核受体 (ROB)，在健康细胞中孕酮高度上调，但在子宫内膜异位症细胞中不表达。 Rorb调节眼和骨骼中祖细胞的分化，以响应RA的梯度， 但在子宫内膜中却没有被注意到。我们的初步数据显示这个孕激素敏感基因 诱导子宫内膜干细胞对黄体酮和维甲酸的反应而分化。我们预测 子宫内膜干细胞中Rorb的缺失会导致病变细胞的分化和存活发生改变。我们 将利用芯片序列技术确定rob的结合部位，并评价rob在体内和体外的作用。 以确定黄体酮和RA信号如何协调体内的生物学变化 子宫内膜异位症。2)明确ER？靶基因在子宫内膜异位症中的生物学作用。厄伯是世界上 受DNA甲基化影响的最重要的靶点，以及其启动子上的甲基化缺失会导致 雌激素受体的病理表达及对雌激素反应的改变。ChlP-seq和基因芯片的初步研究 确定Rerg和SGK1酶是ERç的唯一靶点，并认为这些基因介导了细胞外信号转导。 雌激素反应中的炎症和促生存信号。我们假设异常的表达 这些基因改变雌激素和细胞周期依赖的基因表达和功能。我们将确定 RERG和SGK1参与子宫内膜异位症发病的分子机制 针对这两种酶的治疗将为这种疾病提供替代治疗策略。3)至 确定类固醇激素和RA依赖的DNA甲基化的潜在机制是 子宫内膜异位症病因学基础。我们的初步数据确定了DNA的大规模差异 甲基化后疾病的确立。我们假设甲基化的关键缺陷已经 在注定要发展为子宫内膜异位症的女性的在位内膜中建立或触发。使用高 吞吐量策略来检查单个细胞，这一目标将检查关键基因的甲基化状态，以及 类固醇激素，RA，直接操纵甲基化环境的一组酶 和它们的核受体，并专注于DNMT，它甲基化DNA和Tets，去甲基化 DNA类固醇和类风湿关节炎对全基因组的影响尚未被很好地理解；然而，独特的 子宫内膜异位症的表观遗传学特性表明，对这些酶的全基因组研究将揭示最 触发疾病进展的关键靶点。

Endometriosis, the pathologic endometrium-IIke tissue on pelvic organs, develops and persists in part due to defective apoptosis and is associated with pelvic pain and infertility. Our overall hypothesis is that defective steroid hormone and RA signaling is responsible for the abnormal reprogramming of endomethotic cells. We propose 3 aims. 1) To determine whether RORB deficiency, a consequence of progesterone resistance, is responsible for altered cell survival and differentiation in endometriosis. We uncovered a novel nuclear receptor (RORB), highly upregulated in response to progesterone in healthy cells, but absent in endometriotic cells. RORB regulates the differentiation of progenitor cells in the eye and in the bone in response to RA gradients, but has gone unnoticed in the endometrium. Our preliminary data suggest this progesterone-sensitive gene elicits the differentiation of endometrial stem cells in response to both progesterone and RA. We predict the loss of RORB in endometrial stem cells gives rise to diseased cells with altered differentiation and survival. We will determine the binding sites of RORB using CHIP-seq, and evaluate the in vivo and in vitro roles of RORB in healthy and diseased tissues to ascertain how progesterone and RA signaling coordinate biological changes in endometriosis. 2) To define the biological roles of key ERß target genes in endometriosis. ERß is one of the most significant targets affected by DNA methylation, and the loss of methylation across its promoter results in pathologic expression of ERß and altered response to estrogen. Preliminary ChlP-seq and microarray studies identified the enzymes RERG and SGK1 as unique targets of ERß, and suggested these genes mediate pro- inflammatory and pro-survival signaling in response to estrogen. We hypothesize the abnormal expression of these genes alters estrogen- and cell cycle-dependent gene expression and function. We will determine the molecular mechanism by which RERG and SGK1 contribute to the pathogenesis of endometriosis, as emerging therapies targeting these two enzymes would provide alternative treatment strategies for the disease. 3) To determine the mechanisms underlying steroid hormone and RA-dependent DNA methylation that are fundamental to endometriosis etiology. Our preliminary data identified large-scale differences in DNA methylation after the establishment of the disease. We hypothesize that key defects in methylation are already established or triggered in the eutopic endometrium of women destined to develop endometriosis. Using high throughput strategies to examine individual cells, this aim will examine the methylation status of key genes, and the body of enzymes that directly manipulate the methylation landscape in response to steroid hormones, RA and their nuclear receptors and focus on the DNMTs, which methylate DNA and the TETs, which demethylate DNA. The genome-wide effects of steroids and RA has not been well understood; however the unique epigenetic nature of endometriosis suggests that a genome-wide study of these enzymes will unveil the most critical targets that trigger the progression of the disease.