Actin regulatory proteins regulate alveolar macrophage pro-inflammatory signaling

肌动蛋白调节蛋白调节肺泡巨噬细胞促炎症信号传导

• 批准号: 10065309
• 负责人: Sharon Celeste Morley
• 金额: $ 40.13万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-04-01 至 2021-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10065309
• 关键词: Actins; Adhesions; Adopted; Adult; Adult Respiratory Distress Syndrome; Alveolar; Alveolar Macrophages; Anti-Inflammatory Agents; Award; Bacterial Infections; Biology; Blood; Bundling; CD11 Antigens; Cell Adhesion; Cell surface; Cells; Cellular biology; Chemicals; Communicable Diseases; Complement; Data; Development; Disease; Ensure; Environment; Exhibits; F-Actin; Family; Fluorescence Microscopy; Gases; Goals; Granulocyte-Macrophage Colony-Stimulating Factor; Growth Factor; Hematopoietic; Homeostasis; Host Defense; ITGAX gene; Immunity; Immunology; Impairment; In Vitro; Infection; Inflammasome; Inflammation; Inflammation Mediators; Inflammatory; Inflammatory Response; Inhalation; Integrins; Interleukin-1 beta; Interleukin-4; L-Plastin; Leukocytes; Life; Link; Lung; Lung Inflammation; Lung diseases; Lung infections; Macrophage Activation; Maintenance; Mechanical ventilation; Mechanics; Mediating; Microscopy; Modeling; Molecular; Mus; Neonatal; Organelles; Pathway interactions; Patient-Focused Outcomes; Phagocytes; Pneumonia; Predisposition; Production; Protein Tyrosine Kinase; Proteins; Publishing; Pulmonary Inflammation; Reagent; Reporter; Resolution; Respiratory Tract Infections; Role; Signal Pathway; Signal Transduction; Site; Specific qualifier value; Streptococcus pneumoniae; Stretching; Structure; System; Testing; Therapeutic; Tissues; Translating; Translations; Tyrosine; Universities; Ventilator-induced lung injury; Washington; Work; base; cytokine; experience; genetic regulatory protein; healing; improved; inflammatory milieu; innovation; insight; macrophage; mature animal; mechanical force; mechanotransduction; migration; monocyte; mouse model; neonatal period; novel; novel therapeutic intervention; pathogen; precursor cell; pup; recruit

PROJECT SUMMARY Alveolar macrophages (AMs) are lung-resident phagocytes essential to pulmonary host defense. Perturbation of AM biology contributes to a variety of lung diseases (including acute respiratory distress syndrome and ventilator-induced lung injury) and to susceptibility to respiratory infections. Detailing the molecular mechanisms that control AM activation and development will therefore inform new therapeutic strategies to modulate pulmonary inflammation in multiple diseases. During the course of our initial award, we discovered that mice lacking the actin-bundling protein L-plastin (LPL; LPL-/- mice) were profoundly susceptible to pneumococcal lung infection. We discovered that susceptibility to lung infection correlated with a deficit of AMs, and we confirmed an AM-specific requirement for LPL in bacterial clearance using CD11c.Cre-specific deletion of LPL (newly generated CD11.Cre+-LPLfl/fl mice). We then leveraged our expertise and reagents to probe a novel and significant finding that AM development specifically requires LPL (Blood, 2016). This renewal is built upon published and preliminary data generated while investigating the role of LPL in AM development and function. First, we found that LPL is specifically required for the migration of AM precursor cells (monocytes and/or pre-AMs) into the alveolar space, where the essential growth factor GM-CSF is produced. Second, we found that LPL enables the migration of monocytes and macrophages by supporting podosomes. Podosomes are integrin-mediated, F-actin-based organelles that promote macrophage adhesion and migration. Podosomes also mediate mechanotransduction, translating mechanical force exerted upon the cell into intracellular signaling that alters macrophage biology. Third, we found that AMs from LPL-/- mice exhibited defective IL-1β production after NLRP3 inflammasome activation, and that NLRP3 inflammasome activation was mechanosensitive. Finally, we found that airway administration of exogenous GM-CSF to neonatal LPL-/- mice, during the normal temporal window of AM development, rescued AM numbers and protected adult animals from subsequent bacterial infection. From these findings, we formulated our central hypothesis: LPL, via its function in podosomes, mediates mechanotransduction that regulates macrophage pro-inflammatory signaling. We will test this hypothesis by 1) defining, in unprecedented detail, the formation of podosomes in the presence and absence of LPL; 2) determining which macrophage pro- and anti-inflammatory states require LPL and/or are mechanosensitive; and 3) defining mechanisms by which GM-CSF regulates AM mechanosensitivity and IL-1β production in WT and LPL-deficient AMs to show that AM biology can be therapeutically modified. The combination of the PI's experience in cell biology, immunology and infectious disease, the assembled team of collaborators, and the environment at Washington University ensure that this work will be accomplished. Completion of these studies will illuminate a novel pathway by which alteration in tissue stiffness (compliance) during disease states may alter the inflammatory environment through macrophage mechanotransduction.

项目概要 肺泡巨噬细胞（AM）是肺部驻留的吞噬细胞，对于肺部宿主防御至关重要。 AM 生物学的扰动会导致多种肺部疾病（包括急性呼吸窘迫 综合症和呼吸机引起的肺损伤）以及对呼吸道感染的易感性。 因此，控制 AM 激活和发育的分子机制将为新的治疗提供信息 在我们最初获奖的过程中，我们提出了调节多种疾病的肺部炎症的策略。 发现缺乏肌动蛋白捆绑蛋白 L-plastin 的小鼠（LPL；LPL-/- 小鼠）极易受影响 我们发现对肺炎球菌肺部感染的易感性与缺乏相关。 AMs，我们使用 CD11c.Cre 特异性确认了 LPL 在细菌清除中的 AM 特异性要求 然后，我们利用我们的专业知识和试剂来删除 LPL（新生成的 CD11.Cre+-LPLfl/fl 小鼠）。 探索一项新颖且重要的发现，即 AM 开发特别需要 LPL（Blood，2016）。 此次更新是基于调查 LPL 的作用时生成的出版物和初步数据。 首先，我们发现LPL对于AM前体的迁移是特别需要的。 细胞（单核细胞和/或前 AM）进入肺泡腔，其中必需生长因子 GM-CSF 其次，我们发现 LPL 通过支持单核细胞和巨噬细胞的迁移。 足小体是整合素介导的、基于 F-肌动蛋白的细胞器，可促进巨噬细胞粘附。 足小体还介导机械传导，翻译施加在身体上的机械力。 第三，我们发现来自 LPL-/- 小鼠的 AMs。 NLRP3 炎症小体激活后表现出 IL-1β 产生缺陷，并且 NLRP3 炎症小体 最后，我们发现对新生儿气道施用外源性 GM-CSF 具有机械敏感性。 LPL-/- 小鼠在 AM 发育的正常时间窗口期间，挽救了 AM 数量并保护了成年小鼠 根据这些发现，我们制定了我们的中心假设：LPL， 通过其在足小体中的功能，介导调节巨噬细胞促炎症的机械转导 我们将通过以下方式检验这一假设：1）以前所未有的细节定义足体的形成。 LPL 的存在和缺失；2) 确定哪些巨噬细胞促炎和抗炎状态需要 LPL 和/或具有机械敏感性；以及 3) 确定 GM-CSF 调节 AM 机械敏感性的机制 WT 和 LPL 缺陷 AM 中 IL-1β 的产生表明 AM 生物学可以进行治疗性修改。 结合了 PI 在细胞生物学、免疫学和传染病方面的经验，组建的团队 合作者和华盛顿大学的环境确保了这项工作的完成。 这些研究的完成将阐明一条新的途径，通过该途径改变组织硬度（顺应性） 在疾病状态下，可能会通过巨噬细胞机械转导改变炎症环境。

项目成果

Efficient T Cell Migration and Activation Require L-Plastin.

有效的 T 细胞迁移和激活需要 L-Plastin。

• 发表时间: 2022
• 作者: Joshi, Hemant;Morley, Sharon Celeste
• 通讯作者: Morley, Sharon Celeste

Novel Mouse Model Reveals That Serine Phosphorylation of L-Plastin Is Essential for Effective Splenic Clearance of Pneumococcus.

新型小鼠模型揭示 L-plastin 的丝氨酸磷酸化对于有效的脾脏清除肺炎球菌至关重要。

• 发表时间: 2021
• 作者: Anaya, Edgar P;Lin, Xue;Todd, Elizabeth M;Szasz, Taylor P;Morley, S Celeste
• 通讯作者: Morley, S Celeste

T cells go with the flow: aquaporin 4 is required for full T-cell activation.

T 细胞随波逐流：T 细胞完全激活需要水通道蛋白 4。

• 发表时间: 2023-06-01
• 影响因子: 5.5
• 作者: Morley; Sharon Celeste
• 通讯作者: Sharon Celeste

L-plastin enhances NLRP3 inflammasome assembly and bleomycin-induced lung fibrosis.

L-plastin 增强 NLRP3 炎性体组装和博莱霉素诱导的肺纤维化。

• 发表时间: 2022-03-15
• 影响因子: 8.8
• 作者: Joshi, Hemant;Almgren;Anaya, Edgar P;Todd, Elizabeth M;Van Dyken, Steven J;Seth, Anushree;McIntire, Katherine M;Singamaneni, Srikanth;Sutterwala, Fayyaz;Morley, Sharon C
• 通讯作者: Morley, Sharon C

High-resolution imaging of protein secretion at the single-cell level using plasmon-enhanced FluoroDOT assay.

使用等离子体增强 FluoroDOT 测定在单细胞水平上对蛋白质分泌进行高分辨率成像。

• 发表时间: 2022-08-22
• 作者: Seth, Anushree;Mittal, Ekansh;Luan, Jingyi;Kolla, Samhitha;Mazer, Monty B;Joshi, Hemant;Gupta, Rohit;Rathi, Priya;Wang, Zheyu;Morrissey, Jeremiah J;Ernst, Joel D;Portal;Morley, Sharon Celeste;Philips, Jennifer A;Singamaneni
• 通讯作者: Singamaneni