Micro RNA Regulation of Human Airway Epithelial Phenotype

描述（由申请人提供）： 该申请解决了广泛的挑战领域“（15）转化科学”和特定的挑战主题“15-HL-102：基于microRNA技术开发心脏，肺和血液疾病的新治疗策略”。项目名称为“microRNA Regulation of Human Airway Epithelial Cell Phenotype”。microRNA（miRNAs）在调节基因组和基因中起着关键作用，并且已知参与与肺部疾病相关的多种生物学过程，例如肺发育、肺部炎症、对吸烟的反应和肺癌。然而，大多数miRNA在关键肺细胞类型（如人支气管上皮（hBE）细胞）中的表达和功能仍然知之甚少。当hBE细胞在体外气液界面（ALI）生长时，它们概括了体内天然上皮的结构和功能的关键方面。本项目的目标是全面了解ALI hBE细胞中的miRNA库及其在相关损伤/修复条件下的表达。然后，我们将测试选择，高度调控的miRNA在这一重要细胞类型中的功能。该建议的中心假设是，miRNAs是hBE细胞分化和对损伤的反应的关键调节因子，miRNAs在病理状态下发生改变，并且调节miRNAs可以改变hBE细胞表型以获得治疗益处。为了验证这一假设，我们提出了以下具体目标：目标1）全面描述hBE细胞在正常分化期间和一系列相关损伤/修复条件下的miRNA表达;目标2）确定操纵候选miRNA表达的功能后果hBE细胞。我们将在正常生长和分化过程中以及在一系列高度相关的损伤/修复条件下，在ALI hBE细胞中进行最先进的miRNA发现研究和相关的miRNA和mRNA表达阵列研究。我们将操纵miRNA表达作为概念验证，即调节miRNA可以改变hBE细胞表型以获得治疗益处。这些实验是由资源、专业知识和规模经济的独特组合实现的，并且是雄心勃勃的，但在RFA-OD-09-003的2年范围内可以实现。它们通过减轻细胞类型异质性和采样问题来扩展和补充人类患者样本中的研究。该模型系统将创建一个有价值的数据库，并将能够评估健康和疾病中hBE细胞中的miRNA功能，作为基于miRNA的肺部疾病改进治疗的关键里程碑测试平台。 公共卫生相关性：在这项研究中，我们将全面确定一个关键的肺细胞类型，人支气管上皮细胞的miRNA库，并将测试新的和/或高度调节的miRNA的功能，以改变它们的结构和功能。我们将创建一个有价值的数据库和一个平台，用于对影响超过3000万美国人的肺部疾病的改进治疗进行关键的里程碑式测试。

DESCRIPTION (provided by applicant): This application addresses the broad challenge area "(15) Translational Science" and specific challenge topic, "15-HL-102: Develop new therapeutic strategies for heart, lung, and blood diseases based on microRNA technology". The project title is "microRNA Regulation of Human Airway Epithelial Cell Phenotype". microRNAs (miRNAs) play critical roles regulating genomes and genes and are known to be involved in multiple biological processes relevant to lung diseases, such as lung development, lung inflammation, the response to tobacco smoking and lung cancer. However, the expression and function of the majority of miRNAs in critical lung cell types, such as human bronchial epithelial (hBE) cells, remains poorly understood. When hBE cells are grown in vitro at an air-liquid interface (ALI), they recapitulate key aspects of the structure and function of the native epithelium in vivo. The goals of this project are to gain a comprehensive understanding of the miRNA repertoire in ALI hBE cells and its expression under relevant injury/repair conditions. We will then test the function of select, highly regulated miRNAs in this important cell type. The central hypothesis of this proposal is that miRNAs are critical regulators of hBE cell differentiation and the response to injury, that miRNAs become altered in pathologic states, and that modulating miRNAs can change hBE cell phenotype for therapeutic benefit. To test this hypothesis we pose the following specific aims: Aim 1) Develop a comprehensive portrait of miRNA expression in hBE cells during normal differentiation and in a spectrum of relevant injury/repair conditions; and Aim 2) Determine functional consequences of manipulating expression of candidate miRNAs in hBE cells. We will conduct state-of-the art miRNA discovery studies and correlative miRNA and mRNA expression array studies in ALI hBE cells during normal growth and differentiation and under a series of highly relevant injury/repair conditions. We will manipulate miRNA expression as proof-of-concept that modulating miRNAs can alter hBE cell phenotype for therapeutic benefit. These experiments are enabled by a unique combination of resources, expertise and economies of scale and are ambitious, but achievable within the 2-year scope of RFA-OD-09-003. They extend and complement studies in human patient samples by mitigating cell type heterogeneity and sampling concerns. The model system will create a valuable database and will enable evaluation of miRNA function in hBE cells in health and disease, serving as a platform for critical milestone testing of improved treatments for lung diseases based on miRNA. PUBLIC HEALTH RELEVANCE: In this study we will comprehensively determine the miRNA repertoire of a key lung cell type, human bronchial epithelial cells, and will test the function of novel and/or highly regulated miRNAs to change their structure and function. We will create a valuable database and a platform for critical milestone testing of improved treatments for lung diseases that afflict greater than 30 million Americans.