Translational Regulation in Bronchial Epithelial Cells

支气管上皮细胞的翻译调控

• 批准号: 8683074
• 负责人: Ann-Bin Shyu
• 金额: $ 38万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-25 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8683074
• 关键词: 3' Untranslated Regions; Affect; Air; Airborne Particulate Matter; Allergens; Area; Asthma; Biochemical; Cell Culture Techniques; Cells; Chemicals; Chronic; Code; Cytoplasmic Tail; Development; Disease; Environment; Epithelial; Epithelial Cells; Gene Expression; Homeostasis; Human; Infectious Agent; Inflammation; Inflammation Mediators; Inflammatory; Inflammatory Response; Interleukin-6; Lead; Link; Liquid substance; Maintenance; Mediating; Mediator of activation protein; Messenger RNA; Methodology; MicroRNAs; Molecular; Mutagenesis; Nature; Pathogenesis; Pathway interactions; Play; Porifera; Post-Transcriptional Regulation; Process; Production; Protein Biosynthesis; Proteins; RNA Processing; Regulation; Research; Role; Site; Source; Speed; Stress; Translational Regulation; Translations; airway epithelium; airway inflammation; allergic airway inflammation; asthmatic patient; base; bronchial epithelium; design; in vivo; insight; mouse model; novel; novel therapeutic intervention; repaired; research study; translation factor

DESCRIPTION (provided by applicant): The objective of this proposal is to elucidate the post-transcriptional mechanisms controlling translation and subcellular localization of mRNAs involved in inflammatory responses in bronchial epithelial cells. Epithelial cells of the bronchial airways are directly exposed to the environment and are the first line of defense against airborne particulate matter, allergens and infectious agents. Mounting evidence suggests that bronchial epithelial cells play a pivotal, multifaceted role not only in the maintenance of physico-chemical homoeostasis of the airways but also in the pathogenesis of airway diseases. During allergic airway inflammation, the epithelium is both a source of mediator production as well as a target of remodeling processes. Yet, little is known about post-transcriptional regulation of mediator, effector and remodeling gene expression in these cells during the inflammatory response, and about how this regulation may be altered when airway inflammation becomes persistent, such as in asthma. We have carried out extensive preliminary studies using primary normal or asthmatic human bronchial epithelial (HBE) cells cultured at the liquid-air interface, human bronchial epithelial BEAS-2B cells, and an in vivo mouse model of allergic airway inflammation. The results suggest that bronchial epithelial cells from asthmatics undergo a coordinated alteration of miRNA profile/function and of P- bodies, cytoplasmic domains for storage and/or degradation of translationally repressed mRNAs. These changes lead to a hyperactive state of the protein synthesis that contributes to sustaining excess production of inflammatory mediators and thus perpetuating the chronic nature of asthmatic inflammation. In the proposed study we will: 1) Determine whether a reduction in miR-26 and miR-16 abundance contributes to the persistent, elevated level of IL-6 observed in asthmatic primary HBE cells; 2) Define the role of a group of miRNAs that are significantly down-regulated in asthmatic primary HBE cells in controlling the activity of translation machinery in bronchial epithelial cells; and 3) Determine whether a reduction in P-bodies is a hallmark of activated bronchial epithelial cells, and how alteration of P-body assembly and disassembly influences the inflammatory response in bronchial epithelial cells. The airway epithelium is fundamentally abnormal in asthma, but the molecular and biochemical bases for this abnormality remains largely undefined. We will employ state-of-the-art approaches to gain critical insights into translational regulation of inflammatory mediator production and the pathological mechanism by which this regulation may be altered in bronchial epithelial cells in asthma. The proposed studies involve an unexplored but exciting new area of research in airway inflammation and are likely to introduce novel concepts and methodologies to the field of airway inflammation. The results have a potential for identifying new targets to speed development of new therapeutic approaches to the chronic airway inflammation associated with asthma.

描述（由申请人提供）：本提案的目的是阐明控制支气管上皮细胞炎症反应中涉及的 mRNA 的翻译和亚细胞定位的转录后机制。支气管气道的上皮细胞直接暴露于环境，是抵御空气中颗粒物、过敏原和传染源的第一道防线。越来越多的证据表明，支气管上皮细胞不仅在维持气道物理化学稳态方面发挥着关键的、多方面的作用，而且在气道疾病的发病机制中也发挥着关键的、多方面的作用。在过敏性气道炎症期间，上皮既是介质产生的来源，也是重塑过程的目标。然而，人们对炎症反应过程中这些细胞中介质、效应子和重塑基因表达的转录后调节以及当气道炎症持续存在时（例如哮喘）如何改变这种调节知之甚少。我们使用在液-气界面培养的原代正常或哮喘人支气管上皮 (HBE) 细胞、人支气管上皮 BEAS-2B 细胞以及过敏性气道炎症的体内小鼠模型进行了广泛的初步研究。结果表明，哮喘患者的支气管上皮细胞经历了 miRNA 谱/功能和 P-体、用于储存和/或降解翻译抑制的 mRNA 的细胞质结构域的协调改变。这些变化导致蛋白质合成的过度活跃状态，从而有助于维持炎症介质的过量产生，从而使哮喘炎症长期存在。在拟议的研究中，我们将： 1) 确定 miR-26 和 miR-16 丰度的减少是否会导致哮喘原发性 HBE 细胞中观察到的 IL-6 水平持续升高； 2）明确一组在哮喘原代HBE细胞中显着下调的miRNA在控制支气管上皮细胞翻译机器活性中的作用； 3)确定P体的减少是否是支气管上皮细胞活化的标志，以及P体组装和分解的改变如何影响支气管上皮细胞的炎症反应。哮喘患者的气道上皮本质上是异常的，但这种异常的分子和生化基础在很大程度上仍不清楚。我们将采用最先进的方法来深入了解炎症介质产生的翻译调节以及哮喘支气管上皮细胞中这种调节可能被改变的病理机制。拟议的研究涉及气道炎症的一个未经探索但令人兴奋的新研究领域，并可能为气道炎症领域引入新的概念和方法。这些结果有可能确定新的靶标，以加速开发治疗与哮喘相关的慢性气道炎症的新治疗方法。

项目成果

Analysis of interferon-beta mRNA stability control after poly(I:C) stimulation using RNA metabolic labeling by ethynyluridine.

• DOI: 10.1016/j.bbrc.2012.09.144
• 发表时间: 2012-11-09
• 期刊: BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
• 影响因子: 3.1
• 作者: Abe, Kaito;Ishigami, Tomoaki;Shyu, Ann-Bin;Ohno, Shigeo;Umemura, Satoshi;Yamashita, Akio
• 通讯作者: Yamashita, Akio