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.

项目概要 胃肠道出现严重临床表现的纤维化和炎症性疾病，例如 硬皮病 (SSc) 和克罗恩病 (CD) 在女性中更为常见， 然而，其机制解释了较高的发病率和严重程度。 女性这些疾病的一个共同特征仍然不是很清楚。 我们发现了一种将上皮组织完整性与 RNA 联系起来的机制。 我们已经证明，RNAi 机制、miRNA 调节和结肠上皮细胞行为。 粘附连接是细胞的重要组成部分，它招募微处理器和 RNAi诱导沉默复合物（RISC），RNAi机制的核心组件，以及一组特定的 结肠上皮细胞中 miRNA 和 mRNA 的相互作用通过 E- 成员 PLEKHA7 发生。 钙粘蛋白细胞-细胞粘附复合物缺失导致上皮完整性受损、水平降低。 一组 miRNA 的沉默活性，生长促进剂和上皮细胞的 mRNA 表达增加 我们还发现 PLEKHA7 和连接 RNAi 机制的广泛失调。 我们的初步数据表明，连接 RNAi 的破坏会导致过度表达。 细胞外基质（ECM）和促炎调节剂并促进 ECM 重塑。 我们已经报道过，雌激素可能会通过 Src 激活破坏粘附连接的完整性。 激活的 Src 确实会阻止 RNAi 募集到连接处。 雌激素 – Src 介导的上皮粘附连接相关 RNAi 机制的破坏 miRNA 失调会促进女性 ECM 重塑、炎症和纤维化。 假设有两个具体目标：1）雌激素通过破坏粘附分子促进 ECM 重塑 连接 - 相关 RNAi 机制；2) 粘附连接 - 相关 RNAi 机制的破坏 女性肠道中更常见，并且与较高的 SSc 和 CD 发病率相关，这项研究意义重大，因为。 它可能会确定一种分子机制来解释女性纤维化疾病的严重流行，即 目前缺少的这项工作是创新性的，通过引入局部 RNAi 调控的概念。 这项研究的影响在于它将增进我们对粘附连接处的雌激素的理解。 对妇女健康造成重大负担的疾病的根本机制原因，提供了机会 由于这涉及 miRNA 调节作为该机制的焦点，因此该研究 可以促进基于RNA的疗法的未来发展。 此拟议补充不会导致家长补助金发生变化。