Estrogen-mediated disruption of an E-cadherin - associated RNAi machinery promotes fibrotic diseases in women

雌激素介导的 E-钙粘蛋白相关 RNAi 机制的破坏促进女性纤维化疾病

• 批准号: 10727795
• 负责人: STEPHEN A DUNCAN
• 金额: $ 29.68万
• 依托单位: MEDICAL UNIVERSITY OF SOUTH CAROLINA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-03 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10727795
• 关键词: Adherens Junction; Affect; Agreement; Anti-Inflammatory Agents; Architecture; Biological Assay; Cadherins; Cell-Cell Adhesion; Cellular Structures; Clinical; Collagen; Colon; Colon Carcinoma; Complex; Crohn' s disease; Data; Deposition; Development; Digestive System Disorders; Disease; Disease Progression; E-Cadherin; Epidermal Growth Factor Receptor; Epithelial Cells; Epithelium; Estrogen Receptors; Estrogens; Exhibits; Extracellular Matrix; Female; Fibrosis; Future; Gastrointestinal Diseases; Gastrointestinal tract structure; Growth; Health; High-Throughput RNA Sequencing; Homeostasis; Image; Incidence; Inflammation; Inflammatory; Investigation; Knockout Mice; Link; Liver diseases; Mediating; Messenger RNA; MicroRNAs; Microprocessor; Molecular; Mus; Muscle; Patients; Personal Satisfaction; Population; Prevalence; Proteins; RNA Interference; Regulation; Reporting; Scleroderma; Severities; Severity of illness; South Carolina; Systemic Scleroderma; Testing; Therapeutic Intervention; Tissues; Treatment Cost; Up-Regulation; Woman; Women' s Health; Work; burden of illness; cell behavior; cell motility; cell transformation; cost estimate; estrogen disruption; improved; inflammatory marker; innovation; mRNA Expression; member; mouse model; novel; overexpression; parent grant; promoter; recruit; src-Family Kinases; therapeutic RNA

PROJECT SUMMARY Fibrotic and inflammatory conditions with severe clinical manifestations in the gastrointestinal tract, such as Scleroderma (SSc) and Crohn’s Disease (CD), are significantly more prevalent in women, posing substantial challenges to their health and well-being. However, the mechanisms explaining the higher incidence and severity of these diseases in women are still not well-understood. Compromised epithelial integrity is a common feature of these conditions. We have discovered a mechanism that links epithelial tissue integrity with the RNA interference (RNAi) machinery, miRNA regulation, and colon epithelial cell behavior. We have shown that the adherens junctions, which is an essential architectural component of the cell, recruit the microprocessor and the RNAi-induced silencing complex (RISC), the core components of the RNAi machinery, as well as a specific set of miRNAs and mRNAs, in colon epithelial cells. This interaction occurs through PLEKHA7, a member of the E- cadherin cell-cell adhesion complex. PLEKHA7 loss results in compromised epithelial integrity, decreased levels and silencing activity of a set of miRNAs, increased mRNA expression of growth promoters and epithelial cell transformation. We have also found extensive dysregulation of PLEKHA7 and of the junctional RNAi machinery in colon cancer. Our preliminary data show that disruption of the junctional RNAi results in overexpression of extracellular matrix (ECM) and pro-inflammatory regulators and promotes ECM remodeling. It has been demonstrated that estrogens may disrupt adherens junction integrity through Src activation. We have reported that activated Src indeed opposes RNAi recruitment to the junctions. Based on these findings, we hypothesize that estrogen – Src mediated disruption of the epithelial adherens junction-associated RNAi machinery and miRNA dysregulation promotes ECM remodeling, inflammation, and fibrosis in women. We will examine our hypothesis under two specific Aims: 1) estrogens promote ECM remodeling through disruption of the adherens junction – associated RNAi machinery; 2) disruption of the adherens junction – associated RNAi machinery is more frequent in the female gut and correlates with higher SSc and CD incidence. This study is significant, since it may identify a molecular mechanism explaining the strong prevalence of fibrotic diseases in women, which is currently missing. The proposed work is innovative, by introducing the concept of localized RNAi regulation by estrogens at adherens junctions. The impact of the study is that it will advance our understanding of the underlying mechanistic causes of diseases that pose significant burden on women’s health, offering opportunities for therapeutic intervention. Since this involves miRNA regulation as the focal point of this mechanism, the study can lead to future development of RNA-based therapeutics. There will be no change in the parent grant resulting from this proposed supplement.

项目摘要 在胃肠道中具有严重临床表现的纤维化和炎症条件，例如 硬皮病（SSC）和克罗恩病（CD）在女性中的普遍性明显更高，构成了很大的 他们的健康和福祉面临挑战。但是，解释了较高事件和严重性的机制 在女性中，这些疾病仍然没有得到充分理解。折衷的上皮完整性是一个常见特征 这些条件。我们发现了一种将上皮组织完整性与RNA联系起来的机制 干扰（RNAi）机械，miRNA调节和结肠上皮细胞行为。我们已经证明了 粘附连接是细胞的重要架构组成部分，招募了微处理器和 RNAI诱导的沉默复合物（RISC），RNAi机械的核心组件以及特定组 在结肠上皮细胞中的miRNA和mRNA。这种相互作用通过e-的成员Plekha7发生。 钙粘蛋白细胞 - 细胞粘合剂复合物。 plekha7损失导致上皮完整性损害，提高了水平 以及一组miRNA的沉默活性，增长启动子和上皮细胞的mRNA表达增加 转型。我们还发现了Plekha7和连接RNAi机械的广泛失调 在结肠癌中。我们的初步数据表明，连接性RNAi的破坏导致过表达 细胞外基质（ECM）和促炎性调节剂并促进ECM重塑。它一直 证明进化可能会通过SRC激活破坏粘附的连接完整性。我们报告了 激活的SRC确实反对RNAi招募到交界处。根据这些发现，我们假设 雌激素 - SRC介导的上皮粘附剂连接相关的RNAi机械和 miRNA失调促进女性的ECM重塑，炎症和纤维化。我们将检查我们的 在两个具体目的下的假设：1）通过破坏粘附剂促进ECM重塑 交界处 - 关联的RNAi机械； 2）粘附交界处的破坏 - 相关的RNAi机械是 在雌性肠道中，更常见，与较高的SSC和CD入射相关。这项研究很重要，因为 它可以确定一种分子机制，解释了女性纤维化疾病的强烈流行，这就是 目前缺少。拟议的工作是通过引入局部RNAi调节的概念而创新的 座子连接处的雌激素。该研究的影响是，它将提高我们对 疾病的基本机械原因对妇女的健康造成了严重燃烧，提供了机会 用于治疗干预。由于这涉及miRNA调节作为该机制的焦点，因此该研究 可能导致基于RNA的疗法的未来发展。 该提议的补充剂导致的父母赠款不会改变。