Molecular mechanisms of gap junction promotion of lesion formation in Endometriosis

间隙连接促进子宫内膜异位症病变形成的分子机制

• 批准号: 10490427
• 负责人: BRUCE J NICHOLSON
• 金额: $ 43.58万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-17 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10490427
• 关键词: Adenoviruses; Adhesions; Adhesives; Affect; Animals; Antibodies; Behavior; CD44 gene; Cell Compartmentation; Cells; Characteristics; Connexin 43; Coupling; Development; Diagnostic; Disease; Disease Progression; Disseminated Malignant Neoplasm; Down-Regulation; Endometrial; Endometrium; Environment; Epithelial; Epithelial Cells; Event; Extravasation; Fertility; Fingerprint; Functional disorder; Gap Junctions; Gene Expression; Genes; Genomics; Goals; HMMR gene; Harvest; Hyaluronic Acid; Immunofluorescence Immunologic; Immunohistochemistry; In Vitro; Infection; Invaded; Lead; Lesion; Mediating; Mesothelium; Modification; Molecular; Mus; Neoplasm Metastasis; Pain; Pathogenesis; Pathogenicity; Pathology; Patients; Peritoneal; Peritoneal Mesothelial Cell; Peritoneum; Phenotype; Play; Process; Property; Proteomics; Retrograde Menstruation; Role; Scaffolding Protein; Seeds; Series; Signal Pathway; Signal Transduction; Soil; Stromal Cells; System; Testing; Tissues; Up-Regulation; Woman; base; cell behavior; collaborative environment; endometriosis; epithelial to mesenchymal transition; in vitro Model; in vivo; intercellular communication; mutant; natural Blastocyst Implantation; novel; receptor; response; single-cell RNA sequencing; small hairpin RNA; therapeutic target; transplant model

Invasive cell behavior is a component of normal (blastocyst implantation, extravasation) and pathogenic processes (metastasis, endometriosis), and is initiated by heterotypic contacts between cells. There is strong evidence, particularly in metastatic cancer, that gap junction (GJ) mediated intercellular coupling (GJIC) between the invading cells and target tissue plays a critical role. Yet gap junctions have not been studied in the most common invasive pathogenesis endometriosis. Endometriosis is thought to arise through retrograde menstruation of endometrial tissue to the peritoneum (occurring in most women), but there is active debate as to whether the disease is due to changes in the endometrium (the seed) or a permissive peritoneal environment (the soil). Project 1 demonstrated a striking downregulation of GJ genes in stromal cells (ESCs), and an inverse upregulation in epithelial cells (EECs) that follows the progression of the disease, and could serve as a diagnostic. Here we show that ESCs from endometriosis patients are unique in showing a large induction of GJIC upon interaction with peritoneal mesothelial cells (PMCs). This heterotypic GJIC was critical for the ESC invasiveness by inducing a disruption of the barrier function of the mesothelium. Conversely, PMCs also selectively affect ESCs from endometriosis patients by triggering Cx43 assembly into GJs, and by inducing a Mesenchymal to Epithelial Transition (MET). These results lend strong support the seed hypothesis. The current proposal builds on these results within the collaborative environment of a P01 that will allow a comprehensive analysis of how intercellular interactions in endometriosis lead to lesion formation. This will include understanding: (a) How GJ, and related adhesive genes, are regulated in endometriosis (Project 1); (b) how ESCs and PMCs initially recognize and stably adhere to one another (Project 3), leading to GJ formation, and; (c) how these GJs induce both modification of the PMC barrier function and promote ESC invasive behavior (Project 2). The goals of Project 2 will be achieved through three aims: Aim 1: Establish what functions of Cx43 are important for initiating invasiveness (i.e. GJIC, hemichannels, protein scaffolds) (1.1) and identify the molecular changes induced in PMCs that disrupt barrier function (1.2).

侵袭性细胞行为是正常（囊胚植入、外渗）和致病性的组成部分 过程（转移、子宫内膜异位症），并且是由细胞之间的异型接触启动的。有很强的 有证据表明，特别是在转移性癌症中，间隙连接 (GJ) 介导细胞间偶联 (GJIC) 入侵细胞和靶组织起着至关重要的作用。然而，间隙连接尚未得到充分研究。 常见的发病机制为侵袭性子宫内膜异位症。子宫内膜异位症被认为是通过逆行引起的 月经将子宫内膜组织转移到腹膜（发生在大多数女性身上），但目前存在着激烈的争论 该疾病是否是由于子宫内膜（种子）的变化或允许的腹膜环境引起的 （土壤）。项目 1 证明了基质细胞 (ESC) 中 GJ 基因的显着下调，以及相反的结果 上皮细胞 (EEC) 随疾病进展而上调，可作为诊断依据。 在这里，我们表明，来自子宫内膜异位症患者的 ESC 的独特之处在于，在 与腹膜间皮细胞（PMC）的相互作用。这种异型 GJIC 对于 ESC 侵袭性至关重要 通过诱导间皮屏障功能的破坏。相反，PMC 也有选择地影响 来自子宫内膜异位症患者的 ESC，通过触发 Cx43 组装成 GJ，并诱导间充质 上皮细胞转变（MET）。这些结果有力地支持了种子假说。 当前的提案建立在 P01 协作环境中的这些结果的基础上，该环境将允许 全面分析子宫内膜异位症中细胞间相互作用如何导致病变形成。这将 包括了解： (a) GJ 和相关粘附基因如何在子宫内膜异位症中受到调节（项目 1）； (二) ESC 和 PMC 如何最初认识并稳定地相互粘附（项目 3），从而导致 GJ 形成， 和; (c) 这些 GJ 如何诱导 PMC 屏障功能的改变并促进 ESC 的侵袭行为 （项目2）。项目 2 的目标将通过三个目标来实现： 目标 1：确定 Cx43 的哪些功能对于启动侵袭性很重要（即 GJIC、半通道、 蛋白质支架）（1.1）并确定 PMC 中诱导的破坏屏障功能的分子变化（1.2）。