CFTR and the Immune Response

CFTR 和免疫反应

• 批准号: 8603272
• 负责人: Emanuela Marina Bruscia
• 金额: $ 40.14万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-01-01 至 2015-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8603272
• 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

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

描述（由申请人提供）：肺部感染和强大的炎症反应是囊性纤维化（CF）的主要临床特征，构成了CF患者发病率和死亡率的主要原因。有人提出，异常气道上皮细胞和异常免疫反应会协作并导致严重的慢性肺部疾病。广泛研究了气道上皮对CF炎症反应的贡献，但是对于原代免疫细胞在介导CF肺病中观察到的超反应性中的作用知之甚少。既不知道免疫细胞对肺表型有直接贡献，或者夸张的炎症反应仅是原发性上皮缺陷的继发性。最近的报告表明，CFTR在巨噬细胞和中性粒细胞的正常功能中可能具有重要作用。此外，我们的初步数据表明，巨噬细胞可能在CF气道的高反应性中起重要作用。 CFTR - / - 巨噬细胞的高反应性的基础机制尚不清楚。众所周知，响应LPS的信号转导是由Toll样受体4（TLR4）介导的，该受体样受体4（TLR4）特别结合了LPS。我们发现，与WT巨噬细胞相比，在LPS刺激时，CF巨噬细胞和膜上表达更高量的TLR4。与辅助LPS结合蛋白（包括MD-2和CD14）以及TLR4信号传导的一致性在激活对PA的先天免疫反应中起着重要作用。这些发现表明，免疫细胞直接有助于CF中的夸张的免疫反应。为了研究这一假设，我们提出：i。）研究CFTR+细胞的造血植入是否会改善CFTR - / - 小鼠中的高炎性免疫反应。我们将同时使用体外测定，以及体内研究来确定CFTR无效的免疫细胞在CFTR - / - 小鼠的异常免疫反应中是否起主要作用，以及WT免疫细胞是否可以纠正此反应； ii。）我们将使用基因互补策略来阐述CFTR+上皮细胞和CFTR+巨噬细胞在慢性发炎中对LPS的体内反应中的作用。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小鼠和III相似的炎症反应。 CFTR中的CFTR - / - 细胞

项目成果

Pharmacological modulation of the AKT/microRNA-199a-5p/CAV1 pathway ameliorates cystic fibrosis lung hyper-inflammation.

• DOI: 10.1038/ncomms7221
• 发表时间: 2015-02-10
• 期刊: NATURE COMMUNICATIONS
• 影响因子: 16.6
• 作者: Zhang, Ping-xia;Cheng, Jijun;Zou, Siying;D'Souza, Anthony D.;Koff, Jonathan L.;Lu, Jun;Lee, Patty J.;Krause, Diane S.;Egan, Marie E.;Bruscia, Emanuela M.
• 通讯作者: Bruscia, Emanuela M.

Targeting the Intracellular Environment in Cystic Fibrosis: Restoring Autophagy as a Novel Strategy to Circumvent the CFTR Defect.

• DOI: 10.3389/fphar.2013.00001
• 发表时间: 2013
• 期刊: Frontiers in pharmacology
• 影响因子: 5.6
• 作者: Villella VR;Esposito S;Bruscia EM;Maiuri MC;Raia V;Kroemer G;Maiuri L
• 通讯作者: Maiuri L

Disease-relevant proteostasis regulation of cystic fibrosis transmembrane conductance regulator.

囊性纤维化跨膜电导调节器的疾病相关蛋白质稳态调节。

• DOI: 10.1038/cdd.2013.46
• 发表时间: 2013
• 期刊: Cell death and differentiation
• 影响因子: 12.4
• 作者: Villella,VR;Esposito,S;Bruscia,EM;Vicinanza,M;Cenci,S;Guido,S;Pettoello-Mantovani,M;Carnuccio,R;DeMatteis,MA;Luini,A;Maiuri,MC;Raia,V;Kroemer,G;Maiuri,L
• 通讯作者: Maiuri,L