Epithelial adherens junctions regulate colon cell behavior through RNAi and lncRNAs

上皮粘附连接通过 RNAi 和 lncRNA 调节结肠细胞行为

• 批准号: 10579220
• 负责人: Antonis Kourtidis
• 金额: $ 33.22万
• 依托单位: MEDICAL UNIVERSITY OF SOUTH CAROLINA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-03-26 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10579220
• 关键词: Adherens Junction; Anchorage-Independent Growth; Architecture; Behavior; Biology; Cell-Cell Adhesion; Cells; Cellular Structures; Code; Colon; Colon Carcinoma; Colonic Neoplasms; Colorectal Cancer; Complex; Data; Development; Disease; Disease Progression; E-Cadherin; Epithelial Attachment; Epithelial Cells; Epithelium; Foundations; High Prevalence; Homeostasis; In Vitro; Incidence; Inflammatory Bowel Diseases; Intestinal Cancer; Intestinal Diseases; Knowledge; Link; Maintenance; Malignant Neoplasms; Messenger RNA; MicroRNAs; Microprocessor; Molecular; Oncogenic; Pathway interactions; Patients; Phenotype; Precancerous Conditions; Publishing; RNA; RNA Interference; RNA-Induced Silencing Complex; Regulation; Research; Role; Sampling; Testing; Tissues; Untranslated RNA; Up-Regulation; Work; cell behavior; cell transformation; colon cancer cell line; colon cancer progression; experimental study; gastrointestinal; in vivo; innovation; insight; metaplastic cell transformation; novel; novel diagnostics; novel therapeutic intervention; overexpression; promoter; recruit; therapeutic RNA; tool; transcriptome sequencing; tumor; tumorigenesis; tumorigenic

PROJECT SUMMARY Compromised epithelial integrity is a hallmark of gastrointestinal abnormalities, such as inflammatory bowel disease and colon cancer, which is the third most prevalent and second most lethal form of cancer. The high incidence rates of these diseases suggest that we still don’t fully understand the underlying mechanisms. Recently, we discovered a mechanism that links epithelial tissue integrity with the RNA interference (RNAi) machinery and with miRNA regulation. 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, in colon epithelial cells. This interaction occurs through PLEKHA7, a novel partner of the E-cadherin cell-cell adhesion complex. PLEKHA7 loss results in compromised epithelial integrity, decreased levels and function of a set of miRNAs and in increased anchorage-independent growth, an indicator of epithelial transformation. Our preliminary data show extensive mis-localization or loss of PLEKHA7 in colon cancer cell lines and tumor patient samples. Interestingly, an RNA-CLIP experiment followed by RNA sequencing revealed association of PLEKHA7 with long non-coding RNAs (lncRNAs). LncRNAs can interact with miRNAs and the RNAi machinery in multiple ways and a number of them has been implicated in intestinal diseases. However, our knowledge on lncRNA regulation and function is still limited. Our preliminary data show that PLEKHA7 loss results in altered expression of a number of these lncRNAs, including upregulation of MIR17HG, a known promoter of cellular transformation. We hypothesize that the adherens junctions recruit and regulate the RNAi machinery and lncRNAs to maintain colon epithelial homeostasis. We will test our hypothesis in two Aims that will determine whether: 1) PLEKHA7 suppresses MIR17HG levels through RISC and miRNAs at adherens junctions; 2) PLEKHA7 maintains the normal colon epithelial phenotype through miRNAs and MIR17HG. This study is significant, since it provides a missing mechanistic link between epithelial architecture and cell behavior and a new unexpected localized regulation of RNAi and lncRNAs. The proposed work is innovative, because it tethers two previously unrelated fields, cell-cell adhesion and non-coding RNA biology. The impact of the study is that it will advance our understanding of the underlying mechanistic causes of intestinal diseases and will lay the foundation for the systematic interrogation of the newly discovered connection between the adherens junctions, the RNAi machinery and non-coding RNAs.

项目摘要 折衷的上皮完整性是胃肠道异常的标志，例如炎症肠 疾病和结肠癌，这是癌症的第三大流行和第二大致命形式。高 这些疾病的发病率表明我们仍然不完全了解潜在机制。 最近，我们发现了一种将上皮组织完整性与RNA干扰（RNAI）联系起来的机制 机械和miRNA调节。我们已经证明了粘附连接，这是必不可少的 细胞的建筑成分，募集微处理器和RNAI诱导的沉默复合物（RISC）， 在结肠上皮细胞中，RNAi机械的核心成分以及一组特定的miRNA。这 相互作用通过e-钙粘蛋白细胞 - 细胞粘合剂复合物的新型伴侣Plekha7发生。 Plekha7 损失导致上皮完整性受损，一组miRNA的水平和功能降低和功能 增加了锚固非依赖性的生长，这是上皮转化的指标。我们的初步数据显示 在结肠癌细胞系和肿瘤患者样品中，广泛的失误或损失PLEKHA7。有趣的是， RNA-CLIP实验，然后进行RNA测序揭示了Plekha7与长期非编码的关联 RNA（lncrnas）。 LNCRNA可以通过多种方式与miRNA和RNAi机械相互作用 其中已暗示在肠道疾病中。但是，我们对lncRNA调节和功能的了解 仍然有限。我们的初步数据表明，plekha7损失导致许多这些表达改变 LNCRNA，包括MiR17Hg的上调，MiR17Hg是一种已知的细胞转化启动子。我们假设这一点 粘附连接处招募和调节RNAi机械和LNCRNA，以维持结肠上皮 稳态。我们将以两个确定是否抑制Plekha7的目标来检验我们的假设 MiR17Hg通过RISC和miRNA在粘附连接处的水平； 2）plekha7保持正常结肠 通过miRNA和miR17Hg上皮表型。这项研究很重要，因为它提供了缺失的 上皮结构与细胞行为之间 RNAi和lncrnas。拟议的工作具有创新性，因为它构成了两个以前无关的领域，即细胞细胞 粘附和非编码RNA生物学。该研究的影响是，它将提高我们对 肠道疾病的根本机械原因，并将为系统审讯奠定基础 在粘附连接，RNAi机械和非编码RNA之间的新发现的联系中。