Role of Ezrin in Macrophages

Ezrin 在巨噬细胞中的作用

• 批准号: 10202271
• 负责人: Emanuela Marina Bruscia
• 金额: $ 41.23万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-24 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10202271
• 关键词: Actin-Binding Protein; Actins; Adhesions; Affect; Alveolar; Apoptosis; Asthma; Bacteria; Binding; Biology; Blood Cells; Cause of Death; Cell Shape; Cell membrane; Cell physiology; Cells; Chlorides; Chronic; Clinical; Complex; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; Data; Disease; Event; Extracellular Matrix; F-Actin; Filopodia; Flow Cytometry; Gatekeeping; Gene Mutation; Goals; Health; Human; Image; Immune response; Impairment; In Vitro; Individual; Infection; Inflammation; Inflammatory; Inflammatory Response; Inhalation; Investigation; Knock-out; Knockout Mice; Laboratories; Lipopolysaccharides; Lung; Lung Inflammation; Lung diseases; Lung infections; Macromolecular Complexes; Membrane Proteins; Microscopy; Molecular; Mus; Mutate; Mutation; Nebulizer; PI3K/AKT; Phagocytes; Phagocytosis; Phosphatidylinositol 4,5-Diphosphate; Phosphatidylinositols; Phosphorylation; Physical shape; Population; Production; Protein Family; Proteins; Pseudomonas aeruginosa; Publishing; Pulmonary Cystic Fibrosis; Regulation; Regulator Genes; Reporting; Research; Research Personnel; Resolution; Role; Shapes; Signal Transduction; Spatial Distribution; Staphylococcus aureus; Stimulus; Structure; Structure of parenchyma of lung; TLR4 gene; Techniques; Testing; Tissues; Work; bactericide; base; chemical property; cystic fibrosis patients; cytokine; ezrin; fighting; genetic approach; immunoregulation; in vitro testing; in vivo; in vivo evaluation; inflammatory lung disease; innovation; interstitial; macrophage; member; microorganism; migration; moesin; monocyte; mouse model; preservation; protein degradation; radixin protein; response; small molecule; targeted treatment; therapeutic target; three dimensional cell culture; trafficking

PROJECT SUMMARY Macrophages (MΦs) kill microorganisms, engulf dead cells and debris, and regulate the immune response. They are thus gatekeepers of tissue health, including the lungs. The lung-tissue-resident MΦs (TR-MΦs) are the interstitial and alveolar MΦs, which have complementary but distinct functions. In response to infections, lungs are rapidly populated by waves of Ly6C+ circulating monocytes. In concert with TR-MΦs, these monocytes fight the infection, then facilitate resolution of the inflammatory response. Many chronic lung inflammatory diseases, including cystic fibrosis (CF), are associated with dysregulated MΦ function. Our long- term goal is to understand how different lung MΦ populations contribute to lung hyper-inflammation and infection, and to elucidate the biology of these distinct cell populations. The objective of this proposal is to characterize ezrin’s role in monocyte/MΦ function. Our central hypothesis is that ezrin controls monocyte/MΦ cortical actin organization and signal transduction events in response to inflammatory/infectious stimuli. These cellular changes allow the MΦs to spread, move, phagocytize, and survive, thus shaping the magnitude and quality of the lung immune response to infections. The rationale for these studies is that low ezrin levels have been found in MΦs from patients with CF (our own work). Other investigators have also reported low ezrin levels in blood cells from individuals with asthma. Thus, by elucidating the molecular mechanism by which ezrin shapes lung MΦ functions, we could identify potential therapeutic targets for lung diseases. Our specific aims will test the following hypotheses: (Aim 1) ezrin is necessary for monocyte/MΦ adaptation to the inflamed lung microenvironment; (Aim 2) ezrin is needed for efficient phagocytosis of Staphylococcus aureus and Pseudomonas aeruginosa, two microorganisms that CF patients fail to efficiently eradicate from their lungs; (Aim 3) functional CFTR, the gene that causes CF when mutated, is needed to preserve normal ezrin levels during MΦ activation. The contribution is significant since very little is known about ezrin’s role in regulating lung MΦ activation. Our proposed research is innovative because we will use an unprecedented mouse model in which ezrin is knocked out specifically in monocytes and MΦs. Thus, the proposed studies will investigate in depth the consequences of ezrin loss in monocytes and MΦs during lung infection and inflammation.

项目概要 巨噬细胞 (MΦ) 杀死微生物、吞噬死亡细胞和碎片并调节免疫反应。 因此，它们是组织健康（包括肺部）的守门人。肺组织驻留 MΦ (TR-MΦ) 是 间质和肺泡 MΦ 具有互补但不同的功能。为了应对感染， 肺部迅速充满了 Ly6C+ 循环单核细胞。与 TR-MΦ 配合使用，这些 单核细胞抵抗感染，然后促进炎症反应的消退。许多慢性肺 炎症性疾病，包括囊性纤维化 (CF)，与 MΦ 功能失调有关。我们的长期 术语目标是了解不同的肺 MΦ 群体如何导致肺部过度炎症和 感染，并阐明这些不同细胞群的生物学。该提案的目的是 描述埃兹蛋白在单核细胞/MΦ功能中的作用。我们的中心假设是埃兹蛋白控制单核细胞/MΦ 皮质肌动蛋白组织和信号转导事件响应炎症/感染刺激。这些 细胞变化使 MΦ 能够扩散、移动、吞噬和存活，从而塑造其大小和 肺部对感染的免疫反应的质量。这些研究的基本原理是低埃兹蛋白水平 在 CF 患者的 MΦ 中发现（我们自己的工作）。其他研究人员也报告了低埃兹蛋白 哮喘患者的血细胞水平。因此，通过阐明埃兹蛋白的分子机制 塑造肺 MΦ 功能，我们可以确定肺部疾病的潜在治疗靶点。我们的具体目标 将测试以下假设：（目标 1）ezrin 对于单核细胞/MΦ 适应发炎的肺部是必需的 微环境； （目标 2）埃兹蛋白是金黄色葡萄球菌有效吞噬作用所必需的 铜绿假单胞菌，CF 患者无法有效地从肺部根除的两种微生物； （目标 3）需要功能性 CFTR（突变时导致 CF 的基因）来保持正常的埃兹蛋白水平 在MΦ激活期间。这项贡献意义重大，因为人们对埃兹林在调节中的作用知之甚少。 肺 MΦ 激活。我们提出的研究具有创新性，因为我们将使用前所未有的小鼠模型 其中，ezrin 在单核细胞和 MΦ 中被特异性敲除。因此，拟议的研究将调查 深入了解肺部感染和炎症期间单核细胞和 MΦ 中埃兹蛋白丢失的后果。