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).

侵入性细胞行为是正常（胚泡植入，渗出）和致病性的组成部分 过程（转移，子宫内膜异位症），是通过细胞之间的异型接触引发的。有强大 尤其是在转移性癌症中的证据表明，间隙连接（GJ）介导的细胞间耦合（GJIC） 入侵细胞和靶向组织起着关键作用。然而，尚未最多研究差距 常见的侵入性发病机理子宫内膜异位症。人们认为子宫内膜异位症是通过逆行而产生的 子宫内膜组织到腹膜的月经（在大多数女性中发生），但有主动争论为 疾病是由于子宫内膜（种子）还是允许性腹膜环境引起的 （土壤）。项目1证明了基质细胞中GJ基因的下调（ESC）和一个逆下调 疾病进展的上皮细胞（EEC）的上调，可以用作诊断。 在这里，我们表明，子宫内膜异位症患者的ESC在显示GJIC的大量诱导时是独一无二的 与腹膜间皮细胞（PMC）的相互作用。这种异型GJIC对于ESC的侵入性至关重要 通过诱导间皮的屏障功能的破坏。相反，PMC也有选择地影响 通过将CX43组装触发到GJ中，并通过诱导间质诱导CX43组装来从子宫内膜异位症患者进行ESC 上皮过渡（MET）。这些结果借出了强大的支持种子假设。 当前的建议基于这些结果，在P01的协作环境中，这将允许 对子宫内膜异位中细胞间相互作用如何导致病变形成的全面分析。这会 包括理解：（a）在子宫内膜异位症中如何调节GJ和相关的粘合剂基因（项目1）； （b） ESC和PMC最初如何识别并稳定彼此（项目3），导致GJ组成， 和; （c）这些GJ如何诱导PMC屏障功能的修改并促进ESC侵入性行为 （项目2）。项目2的目标将通过三个目标实现： 目标1：确定CX43的功能对于发起侵入性很重要（即GJIC，Hemichannels， 蛋白质支架）（1.1），并确定破坏屏障功能的PMC中诱导的分子变化（1.2）。