CFTR and the Immune Response

CFTR 和免疫反应

基本信息

批准号：
8010832
负责人：
Emanuela Marina Bruscia
金额：
$ 41.38万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-01-01 至 2014-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8010832
关键词：
Abnormal Epithelial Cell Accounting Acute Affect Alveolar Macrophages Binding Bronchoalveolar Lavage Fluid CD14 gene Cells Chronic lung disease Clinical Clinical Management Cystic Fibrosis Cystic Fibrosis Transmembrane Conductance Regulator Cytokine Signaling Data Defect Disease Engraftment Epithelial Epithelial Cells Goals Health Hematopoietic Human Immune Immune response In Vitro Inbred CFTR Mice Infection Infiltration Inflammatory Inflammatory Response Kinetics Lipopolysaccharides Lung Mediating Membrane Morbidity - disease rate Mus Myeloid Cells Phenotype Play Pseudomonas aeruginosa Pulmonary Cystic Fibrosis Reporting Resolution Role Secondary to Signal Transduction Stimulus Testing Time airway epithelium cystic fibrosis airway cystic fibrosis mouse cystic fibrosis patients cytokine gene complementation improved in vitro Assay in vivo insight lipopolysaccharide-binding protein macrophage mortality neutrophil receptor response toll-like receptor 4 trafficking

项目摘要

DESCRIPTION (provided by applicant): Pulmonary infection and a robust inflammatory response are dominant clinical features of cystic fibrosis (CF), and comprise the major cause of morbidity and mortality in CF patients. It has been suggested that abnormal airway epithelial cells and abnormal immune responses collaborate and result in severe chronic lung disease. The contribution of airway epithelia to the inflammatory response in CF is being extensively studied, however much less is known about the role of primary immune cells in mediating the hyper- responsiveness observed in CF lung disease. Neither is it known if immune cells have a direct contribution to the lung phenotype or if the exaggerated inflammatory response is merely secondary to the primary epithelial defect. Recent reports suggest that CFTR may have an important role in the normal function of both macrophages and neutrophils. Additionally, our preliminary data suggest that the macrophage may play an important role in the hyper-responsiveness of the CF airway. The mechanism(s) underlying the hyper-responsiveness of CFTR-/- macrophages is not known. It is known that signal transduction in response to LPS is mediated by Toll-like receptor 4 (TLR4), which binds to LPS specifically. We have found that upon LPS stimulation, CF macrophages and express higher amounts of TLR4 on their membrane when compared to WT macrophages. In concert with accessory LPS-binding proteins including MD-2 and CD14, and TLR4 signaling plays a major role in the activation of the innate immune response to PA. These findings suggest that immune cells directly contribute to the exaggerated immune response in CF. In order to investigate this hypothesis we propose: i.) to investigate if hematopoietic engraftment of CFTR+ cells will ameliorate the hyper-inflammatory immune response in CFTR-/- mice. We will use both in vitro assays, as well as, in vivo studies to determine if CFTR null immune cells play a primary role in the abnormal immune response in CFTR-/- mice and whether WT immune cells can rectify this response; ii.) we will dissect the roles of CFTR+ epithelial cells and CFTR+ macrophages in the in vivo response to LPS in the chronically inflamed lung using a gene complementation strategy. We will determine if CFTR-/- mice with macrophage specific CFTR expression versus epithelial-cell specific CFTR expression has an inflammatory response that is similar to CFTR-/-, CFTR-/+ or WT mice and iii.) lastly, we will examine if the lack of CFTR is directly responsible for the exaggerated immune response to LPS in macrophages by blocking CFTR in WT cells and by enhancing expression of CFTR in CFTR-/- cells PUBLIC HEALTH RELEVANCE: The ultimate goal of this project is to apply insights gained from these studies to improve the clinical management of people with cystic fibrosis and develop new therapies to treat this disease.

描述（由申请人提供）：肺部感染和强烈的炎症反应是囊性纤维化（CF）的主要临床特征，也是囊性纤维化患者发病和死亡的主要原因。有人认为，异常的气道上皮细胞和异常的免疫反应共同作用，导致严重的慢性肺部疾病。气道上皮细胞对 CF 炎症反应的贡献正在被广泛研究，然而，对于初级免疫细胞在介导 CF 肺病中观察到的高反应性中的作用知之甚少。也不知道免疫细胞是否对肺表型有直接贡献，或者过度的炎症反应是否只是继发于原发性上皮缺陷。最近的报告表明，CFTR 可能在巨噬细胞和中性粒细胞的正常功能中发挥重要作用。此外，我们的初步数据表明巨噬细胞可能在 CF 气道的高反应性中发挥重要作用。 CFTR-/-巨噬细胞高反应性的机制尚不清楚。众所周知，响应 LPS 的信号转导是由 Toll 样受体 4 (TLR4) 介导的，该受体与 LPS 特异性结合。我们发现，与 WT 巨噬细胞相比，在 LPS 刺激后，CF 巨噬细胞在其膜上表达更高含量的 TLR4。与辅助 LPS 结合蛋白（包括 MD-2 和 CD14）相配合，TLR4 信号传导在激活 PA 的先天免疫反应中发挥着重要作用。这些发现表明，免疫细胞直接导致 CF 中过度的免疫反应。为了研究这一假设，我们建议：i.) 研究 CFTR+ 细胞的造血植入是否会改善 CFTR-/- 小鼠的高炎症免疫反应。我们将使用体外测定和体内研究来确定 CFTR 无效免疫细胞是否在 CFTR-/- 小鼠的异常免疫反应中起主要作用，以及 WT 免疫细胞是否可以纠正这种反应； ii.) 我们将使用基因互补策略剖析 CFTR+ 上皮细胞和 CFTR+ 巨噬细胞在慢性炎症肺部 LPS 体内反应中的作用。我们将确定具有巨噬细胞特异性 CFTR 表达与上皮细胞特异性 CFTR 表达的 CFTR-/- 小鼠是否具有与 CFTR-/-、CFTR-/+ 或 WT 小鼠相似的炎症反应，并且 iii.) 最后，我们将检查如果 CFTR 的缺乏直接导致巨噬细胞中对 LPS 的过度免疫反应，则通过阻断 WT 细胞中的 CFTR 和增强 CFTR-/- 细胞中 CFTR 的表达公共健康相关性：该项目的最终目标是应用从这些研究中获得的见解来改善囊性纤维化患者的临床管理并开发治疗这种疾病的新疗法。