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，外渗）和致病的组成部分 过程（转移，子宫内膜异位症），并启动细胞之间的异型接触。有强有力 有证据表明，特别是在转移性癌症中，间隙连接（GJ）介导了细胞间偶联（GJIC）， 侵入的细胞和靶组织起着关键作用。然而，在大多数情况下，缝隙连接还没有被研究。 常见的侵袭性发病机制子宫内膜异位症。子宫内膜异位症被认为是通过逆行 月经的子宫内膜组织的腹膜（发生在大多数妇女），但有积极的辩论， 疾病是由于子宫内膜（种子）的变化还是由于允许的腹膜环境 (the土壤）。项目1表明，基质细胞（ESC）中GJ基因显著下调，而基质细胞中GJ基因的表达则相反。 上皮细胞（EECs）中的上调，其跟随疾病的进展，并且可以用作诊断。 在这里，我们发现子宫内膜异位症患者的ESCs是独特的，在子宫内膜异位症发生后， 与腹膜间皮细胞（PMC）的相互作用。这种异型GJIC对于ESC的侵袭性至关重要 通过诱导间皮细胞屏障功能的破坏。相反，PMC也选择性地影响 通过触发Cx43组装成GJ，并通过诱导间充质干细胞， 上皮转化（MET）。这些结果有力地支持了种子假说。 目前的建议建立在P01协作环境中的这些结果的基础上， 综合分析子宫内膜异位症中细胞间的相互作用如何导致病变形成。这将 包括理解：（a）GJ和相关粘附基因在子宫内膜异位症中如何调节（项目1）;（B） ESC和PMC最初如何识别并稳定地相互粘附（项目3），导致GJ形成， 以及（c）这些GJ如何诱导PMC屏障功能的改变和促进ESC侵袭行为 （项目2）。项目2的目标将通过三个目标实现： 目的1：确定Cx43的哪些功能对于启动侵袭性是重要的（即GJIC，半通道， 蛋白质支架）（1.1），并鉴定破坏屏障功能的PMC中诱导的分子变化（1.2）。