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，其发展和持续部分是由于 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 类固醇激素和RA信号转导是子宫内膜细胞异常重编程的原因。我们 提出三个目标。1)为了确定是否RORB缺乏症，孕激素抵抗的结果， 负责改变子宫内膜异位症中的细胞存活和分化。我们发现了一种新的核受体 （RORB），在健康细胞中响应于孕酮而高度上调，但在增生细胞中不存在。 RORB响应RA梯度调节眼和骨中祖细胞的分化， 但在子宫内膜中没有被注意到。我们的初步数据表明这个对胆固醇敏感的基因 促进子宫内膜干细胞对孕酮和RA的反应。我们预计明年的 子宫内膜干细胞中RORB的缺失导致分化和存活改变的病变细胞。我们 将使用CHIP-seq确定RORB的结合位点，并评估RORB在体内和体外的作用。 健康和患病组织，以确定孕酮和RA信号传导如何协调生物学变化， 子宫内膜异位症2)目的：明确子宫内膜异位症中关键ER β靶基因的生物学作用。Erdom是其中一个 受DNA甲基化影响的最重要的靶点，其启动子的甲基化缺失导致 ER β的病理表达和对雌激素反应的改变。初步ChIP-seq和微阵列研究 确定了酶RERG和SGK 1作为ERK的独特靶点，并表明这些基因介导了 炎症和促存活信号传导。我们推测， 这些基因改变雌激素和细胞周期依赖性基因表达和功能。康贝特人将以 RERG和SGK 1参与子宫内膜异位症发病机制的分子机制， 以这两种酶为靶点的疗法将为该疾病提供替代的治疗策略。3)到 确定类固醇激素和RA依赖的DNA甲基化的潜在机制， 子宫内膜异位症病因学的基础。我们的初步数据表明， 在疾病发生后的甲基化。我们假设甲基化中的关键缺陷已经存在， 在注定发展子宫内膜异位症的妇女的在位子宫内膜中建立或触发。使用高 通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， 直接操纵甲基化景观的酶体，以响应类固醇激素，RA 以及它们的核受体，并将重点放在使DNA甲基化的DNMT和使DNA去甲基化的TMT上。 DNA.类固醇和RA的全基因组效应尚未得到很好的理解;然而， 子宫内膜异位症的表观遗传性质表明，对这些酶的全基因组研究将揭示子宫内膜异位症的最重要的 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.