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.

项目概要 上皮完整性受损是胃肠道异常的标志，例如炎症性肠病 疾病和结肠癌，这是第三大流行和第二大致命的癌症形式。 这些疾病的发病率表明我们仍未完全了解其潜在机制。 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 我们已经证明了粘附连接，这是一个重要的机制。 细胞的结构组成部分，招募微处理器和 RNAi 诱导的沉默复合物 (RISC)， 结肠上皮细胞中 RNAi 机制的核心组件以及一组特定的 miRNA。 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型伙伴。 损失会导致上皮完整性受损、一组 miRNA 的水平和功能下降，并导致 不依赖贴壁的生长增加，这是上皮转化的一个指标。 结肠癌细胞系和肿瘤患者样本中 PLEKHA7 的广泛错误定位或丢失。 RNA-CLIP 实验和 RNA 测序揭示了 PLEKHA7 与长非编码的关联 RNA (lncRNA) 可以通过多种方式与 miRNA 和 RNAi 机制相互作用。 然而，我们对 lncRNA 调控和功能的了解还不够。 我们的初步数据表明 PLEKHA7 丢失会导致其中许多的表达发生改变。 lncRNA，包括 MIR17HG（一种已知的细胞转化启动子）的上调。 粘附连接招募并调节 RNAi 机制和 lncRNA 以维持结肠上皮 我们将在两个目标中检验我们的假设，以确定是否：1) PLEKHA7 抑制。 MIR17HG 通过粘附连接处的 RISC 和 miRNA 表达；2) PLEKHA7 维持正常结肠 这项研究很重要，因为它提供了一个缺失的信息。 上皮结构和细胞行为之间的机制联系以及新的意想不到的局部调节 RNAi 和 lncRNA 是一项创新工作，因为它将两个以前不相关的领域联系在一起，即细胞-细胞。 这项研究的影响在于它将增进我们对粘附和非编码RNA生物学的理解。 肠道疾病的根本机制原因，将为系统研究奠定基础 新发现的粘附连接、RNAi 机制和非编码 RNA 之间的联系。