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: 10661068
• 负责人: BRUCE J NICHOLSON
• 金额: $ 43.64万
• 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/10661068
• 关键词: Adenoviruses; Adhesions; Adhesives; Affect; Animals; Antibodies; Behavior; CD44 gene; Cell Compartmentation; Cell Separation; Cells; Characteristics; Connexin 43; Coupling; Development; Diagnostic; Disease; Disease Progression; Disseminated Malignant Neoplasm; Down-Regulation; Endometrial; Endometrium; Environment; Epithelial Cells; Epithelium; Event; Extravasation; Fertility; Fingerprint; Functional disorder; Gap Junctions; Gene Expression; Genes; Genomics; Germ Cells; 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; Series; Signal Pathway; Signal Transduction; Soil; Stromal Cells; System; Testing; Tissues; Up-Regulation; Woman; cell behavior; collaborative environment; endometriosis; epithelial to mesenchymal transition; in vitro Model; in vivo; intercellular communication; mutant; natural Blastocyst Implantation; novel; permissiveness; 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(胚泡着床、渗出)和致病的一个组成部分 过程(转移、子宫内膜异位症)，并由细胞之间的异型接触启动。有很强的 有证据表明，尤其是在转移性癌症中，缝隙连接(GJ)介导的细胞间偶联(GJIC) 侵袭细胞和靶组织起着至关重要的作用。然而，对缝隙连接的研究还很少。 子宫内膜异位症的常见侵袭性发病机制。子宫内膜异位症被认为是通过逆行发生的 月经的子宫内膜组织到腹膜(发生在大多数妇女)，但有活跃的争论，如 这种疾病是由于子宫内膜(种子)的改变，还是由于腹膜环境的允许 (土壤)。项目1展示了基质细胞(ESCs)中GJ基因的显著下调，以及相反的 随着疾病的发展，上皮细胞(EECS)表达上调，可作为诊断。 在这里，我们显示了来自子宫内膜异位症患者的胚胎干细胞是独一无二的，在子宫内膜异位症患者身上显示出大量的GJIC诱导。 与腹膜间皮细胞的相互作用。这种异型GJIC是ESC侵袭性的关键 通过诱导间皮细胞屏障功能的破坏。相反，私营军事公司也选择性地影响 子宫内膜异位症患者的ESCs通过触发Cx43组装到GJS，并通过诱导间充质细胞 上皮转化(MET)。这些结果有力地支持了种子假说。 目前的提案建立在P01的协作环境中的这些结果之上，该环境将允许 子宫内膜异位症中细胞间相互作用如何导致病变形成的综合分析。这将是 包括了解：(A)GJ和相关黏附基因在子宫内膜异位症中是如何调节的(项目1)；(B) ESCs和PMC最初如何识别并稳定地相互粘合(项目3)，导致GJ的形成， 和；(C)这些GJ如何导致PMC屏障功能的改变和促进ESC的侵袭行为 (项目2)。项目2的目标将通过三个目标实现： 目标1：确定Cx43的哪些功能对启动侵袭(即GJIC，半突， 蛋白质支架)(1.1)，并确定在PMCs中诱导的破坏屏障功能的分子变化(1.2)。