Regulation of Endometriotic Lesion Development by NOTCH1

项目摘要 子宫内膜异位症是一种慢性雌激素依赖型妇科疾病，影响10%-15%的女性 生育年龄导致盆腔疼痛和不孕。它的特点是子宫内膜组织的存在。 子宫外，主要在盆腔腹膜内。子宫内膜异位病变或植入物的活动性，以及 继发性粘连的发展，可能是本病症状的原因。病变发展 很难在患有子宫内膜异位症的妇女中进行研究，因为诊断的严重延误。在过去20年里 多年来，我们一直将狒狒作为适当的模型来更好地检查机构和 子宫内膜异位症病变的进展。理解调节发展和发展的机制 发病时异位病变的进展将为靶向治疗带来新的机会 预防和/或治疗子宫内膜异位症。Notch家族的跨膜受体(NOTCH1-4)转导 负责细胞存活、细胞间通讯和分化的细胞外信号。我们的中环 假说是炎性腹膜环境中存在的IL-6诱导了 E蛋白转录因子E2a和Heb通过直接上调NOTCH1的表达 与NOTCH1启动子结合。NOTCH1的诱导通过以下途径促进子宫内膜异位病变的发展 控制细胞增殖、侵袭和EMT。在这项计划中，我们将利用我们创新的工程鼠标 模型，已建立的狒狒子宫内膜异位症模型以及具有良好特征的人体组织 了解NOTCH1是如何诱导的，以及NOTCH1的诱导和上调如何有助于 子宫内膜异位病变的发生。具体目标1将集中在IL-6在体内的分子机制 子宫内膜异位症背景下，调节E蛋白转录因子(E2a和Heb)的表达诱导 NOTCH1的体外和体内实验，并确定NOTCH1激活信号的分子机制 促进子宫内膜异位症病变发展的途径。具体目标2将侧重于功能研究 使用3D和类器官细胞培养模型和我们独特的荧光标记NOTCH1功能和 功能丧失小鼠模型的分子机制和病理后果的解剖 NOTCH1在子宫内膜异位症组织中的表达上调调节细胞增殖并诱导EMT增强 移民和入侵。《特定目标3》将同时使用转基因小鼠模型和狒狒子宫内膜异位症模型 靶向递送显性负向神经支配样多肽模型(SAHM1) 荧光标记纳米颗粒抑制NOTCH1信号转导作为一种新的治疗方法 与疾病相关的病理过程。这些研究的结果将加强我们的 了解子宫内膜异位症病变的病因学和病理生理学发展并阐明新的观点 将转化为寻找新的非激素治疗方法的途径 子宫内膜异位症。

Project Summary Endometriosis is a chronic, estrogen-dependent gynecological disorder affecting 10-15% of women of reproductive age resulting in pelvic pain and infertility. It is characterized by the presence of endometrial tissue outside the uterus, predominantly in the pelvic peritoneum. The activity of endometriotic lesions or implants, and the development of subsequent adhesions, is the likely causes of the disease symptoms. Lesion development is difficult to study in women with endometriosis because of the significant delays in diagnosis. In the past 20 years we have developed the baboon as an appropriate model to better examine the establishment and progression of endometriotic lesions. Understanding the mechanisms that regulate the development and progression of ectopic lesions at the onset of the disease will result in new opportunities for targeted therapies to prevent and/or treat endometriosis. The Notch family of transmembrane receptors (NOTCH1-4) transduces extracellular signals responsible for cell survival, cell-to-cell communication, and differentiation. Our Central Hypothesis is that IL-6, which is present in the inflammatory peritoneal environment induces the expression of the E-protein transcription factors E2A and HEB, which in turn upregulates NOTCH1 expression by directly binding to the NOTCH1 promoter. The induction of NOTCH1 promotes endometriotic lesion development by controlling cell proliferation, invasion and EMT. In this proposal we will utilize our innovative engineered mouse models, the established baboon endometriosis model together with well characterized human tissues to understand how NOTCH1 is induced and how the induction and upregulation of NOTCH1 contributes to endometriotic lesion development. Specific Aim 1 will focus on the molecular mechanisms by which IL-6, in the context of endometriosis, regulates the expression of E-protein transcription factors (E2A and HEB) to induce NOTCH1 in vitro and in vivo and determine the molecular mechanisms by which NOTCH1 activates signaling pathways that contribute to endometriotic lesion development. Specific Aim 2 will focus on functional studies using 3-D and organoid cell culture models and our unique fluorescence-tagged NOTCH1 gain of function and loss of function mouse models to dissect the molecular mechanisms and pathological consequences by which increased NOTCH1 expression in endometriotic tissues regulates cell proliferation and induces EMT to enhance migration and invasion. Specific Aim 3 will use both engineered mouse models and the baboon endometriosis model for targeted delivery of a dominant negative Mastermind-like (DN-MAML) peptide (SAHM1) using fluorescently labeled nanoparticles to inhibit NOTCH1 signaling as a novel therapeutic approach to inhibit the pathologic processes associated with the disease. The outcomes of these studies will enhance our understanding of the etiology and pathophysiology of endometriotic lesion development and elucidate novel pathways which will translate into identifying novel non-hormonal therapeutic approaches for treating endometriosis.