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 受损的上皮完整性是胃肠道异常的标志，例如炎症性肠 疾病和结肠癌，结肠癌是第三种最普遍和第二种最致命的癌症。高 这些疾病的发病率表明，我们仍然没有完全了解其潜在的机制。 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 与miRNA调控的关系。我们已经表明，粘附连接，这是一个重要的 细胞的结构组成部分，募集微处理器和RNAi诱导的沉默复合物（RISC）， RNAi机制的核心成分，以及一组特定的miRNA，在结肠上皮细胞中。这 相互作用通过PLEKHA7发生，PLEKHA7是E-钙粘蛋白细胞-细胞粘附复合物的新伴侣。PLEKHA7 缺失导致上皮完整性受损，一组miRNA的水平和功能降低， 增加的锚定非依赖性生长，这是上皮转化的指标。我们的初步数据显示 PLEKHA7在结肠癌细胞系和肿瘤患者样品中的广泛错误定位或丢失。有趣的是， RNA测序后的RNA-CLIP实验揭示了PLEKHA7与长非编码区的关联， RNA（lncRNA）。LncRNA可以以多种方式与miRNA和RNAi机制相互作用， 与肠道疾病有关。然而，我们对lncRNA调控和功能的了解 仍然有限。我们的初步数据表明，PLEKHA7缺失导致许多这些基因的表达改变， lncRNA，包括MIR17HG的上调，MIR17HG是细胞转化的已知启动子。我们假设 粘附连接募集并调节RNAi机制和lncRNA，以维持结肠上皮细胞 体内平衡我们将在两个目标中测试我们的假设，以确定是否：1）PLEKHA7抑制 通过粘附连接处的RISC和miRNA的MIR17 HG水平; 2）PLEKHA7维持正常结肠 上皮细胞表型通过miRNA和MIR17HG。这项研究意义重大，因为它提供了一个缺失的 上皮结构和细胞行为之间的机械联系以及新的意想不到的局部调节 RNAi和lncRNA。这项提议的工作是创新的，因为它将两个以前不相关的领域联系在一起， 粘附和非编码RNA生物学。这项研究的影响是，它将促进我们对 肠道疾病的潜在机制的原因，并将奠定基础，系统的审讯 新发现的粘附连接、RNAi机制和非编码RNA之间的联系。