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

项目总结 胃肠道有严重临床表现的纤维化和炎症情况，如 硬皮病(SSC)和克罗恩病(CD)在女性中明显更常见，构成 对他们的健康和福祉的挑战。然而，解释发病率和严重性较高的机制 这些疾病在妇女中的作用仍然不是很清楚。上皮完整性受损是一种常见特征 这些条件。我们已经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)机制，miRNA调节，和结肠上皮细胞行为。我们已经证明了 附着连接，这是细胞的基本架构组件，招募微处理器和 RNAi诱导沉默复合体(RISC)，RNAi机制的核心组件，以及特定的一组 在结肠上皮细胞中的miRNAs和mRNAs。这种相互作用是通过PLEKHA7发生的，它是E- 钙粘附素细胞-细胞黏附复合体。PLEKHA7缺失会导致上皮完整性受损，水平下降 以及一组miRNAs的沉默活性，增加生长促进剂和上皮细胞的mRNA表达 转型。我们还发现了PLEKHA7和连接RNAi机制的广泛失调 结肠癌。我们的初步数据显示，连接RNAi的破坏导致了 细胞外基质(ECM)和促炎调节剂，并促进ECM重塑。一直以来 表明雌激素可能通过激活Src来破坏粘连连接的完整性。我们已经报道了 激活的Src确实反对RNAi在交界处的招募。基于这些发现，我们假设 雌激素-Src介导的上皮细胞破坏与连接相关的RNAi机制和 MiRNA失调促进女性ECM重塑、炎症和纤维化。我们将检查我们的 两个特定目标下的假说：1)雌激素通过破坏粘附物促进ECM重塑 连接相关RNAi机制；2)贴壁连接相关RNAi机制的中断是 更多发生在女性肠道，并与较高的SSC和CD发生率相关。这项研究意义重大，因为 它可能确定一种分子机制，解释女性纤维性疾病的强烈流行，这是 目前失踪。拟议的工作是创新的，通过引入本地化RNAi调控的概念 粘连结合处的雌激素。这项研究的影响是，它将促进我们对 对妇女健康构成重大负担的疾病的根本机制原因，提供了机会 进行治疗干预。由于这涉及到miRNA调控作为这一机制的焦点，因此该研究 可能会导致基于RNA的疗法的未来发展。 这项拟议的补助金不会对父母补助金产生任何影响。

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.