REGULATION OF THE MACROPHAGE INFLAMMATORY PHENOTYPE IN ARDS

ARDS 中巨噬细胞炎症表型的调节

• 批准号: 10094230
• 负责人: John W Christman
• 金额: $ 56.37万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-01-01 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10094230
• 关键词: ADP ribosylation; Ablation; Acute Lung Injury; Address; Adoptive Transfer; Adult Respiratory Distress Syndrome; Affect; Affinity; Alveolar; Alveolar Macrophages; Antibiotics; Apoptosis; Attenuated; Binding; Bleomycin; Blood capillaries; Bone Marrow; Calcineurin; Caring; Cells; Chimera organism; Clinical; Critical Illness; Data; Development; Endothelium; Family; Functional disorder; Gene Expression; Generations; Genetic; Health; Human; Hypoxemia; Inflammation; Inflammatory; Intensive Care Units; Knockout Mice; Laboratories; Life; Ligands; Liquid substance; Lung; Measures; Mediating; Mediator of activation protein; Methods; Molecular; Molecular Target; Morbidity - disease rate; Mus; Outcome; Oxygen; Pathogenesis; Patient-Focused Outcomes; Peptides; Permeability; Pharmacological Treatment; Pharmacology; Phenotype; Pneumonia; Prevention approach; Production; Proteolysis; Public Health; Pulmonary Edema; Pulmonary Inflammation; Recovery; Regulation; Research; Role; TNF gene; TNF-related apoptosis-inducing ligand; Therapeutic Agents; Ventilator; base; cecal ligation puncture; clinically relevant; cytokine; design; experimental study; high risk; improved; improved outcome; inflammatory lung disease; inhibitor/antagonist; macrophage; member; mortality; mouse model; neutrophil; novel; novel strategies; nuclear factors of activated T-cells; patient population; pre-clinical; preclinical efficacy; prevent; promoter; pulmonary function; reconstitution; recruit; response; therapeutically effective; transcription factor; transcription factor NF-AT c3; ventilation

PROJECT SUMMARY Acute Lung Injury (ALI) and Acute Respiratory Distress Syndrome (ARDS) are neutrophilic inflammatory lung diseases with unacceptably high rates of morbidity and mortality. Our supportive data indicate that genetic ablation and pharmacologic inhibition of NFATc3 is beneficial in terms of tighter pulmonary barrier function, decreased cytokine release and neutrophilic inflammation, improved arterial oxygenation, and better survival in mouse models of ARDS/ALI. Based on these and other clinically relevant supportive data, we posit that NFATc3 activation in macrophages induces inflammatory gene expression that mediates the initiation, intensity and duration of ALI/ARDS. We propose three specific aims to address this hypothesis: Specific Aim 1: To identify the mechanism(s) by which NFATc3 deletion protects against ALI/ARDS. We will employ genetically defined mice that globally lack NFATc3 subjected to a well-defined murine model of ALI/ARDS. We will also utilize established methods of NFATc3 KO/WT bone marrow chimera mice, macrophage passive adoptive transfer of lung macrophages, and we will construct novel conditional macrophage selective NFATc3-/- mice to identify the impact of macrophage NFATc3 deficiency on the generation of pulmonary edema, arterial oxygen saturation, and mortality. Specific Aim 2: To determine the mechanisms by which macrophage NFATc3 activation contributes to the development of permeability pulmonary edema. In order to determine how macrophages contribute to the pulmonary edema and severe hypoxemia in ALI/ARDS, we propose to examine the role of NFATc3 regulation of macrophage expression of TNF-related apoptosis-inducing ligand (TRAIL), which inhibits alveolar fluid clearance (AFC), in addition to lung macrophage production of mediators that disrupt barrier function of pulmonary microvascular endothelium. Specific Aim 3: To determine the efficacy of a novel cell permeable peptide NFAT inhibitor in protecting against ALI/ARDS-like pathophysiology in mouse models. Our group has developed a novel NFAT inhibitor, CP9-ZIZIT, that has an impressively low Kd value of 2.6 nM, better cell permeability, higher binding affinity, and improved stability against proteolysis than other available agents. We expect to show that treatment with CP9-ZIZIT alone or in combination with a PARP1 inhibitor will attenuate pulmonary inflammation and alveolar capillary barrier disruption. We expect that this research will provide valuable preclinical data that blocking NFATc3 activation in macrophages is an effective therapeutic agent for preventing ARDS in a high-risk patient population and improving the clinical outcome of patients suffering from moderate and severe ARDS/ALI.

项目摘要 急性肺损伤（Acute lung injury，ALI）和急性呼吸窘迫综合征（acute respiratory Distress Syndrome，ARDS）是一种嗜酸性的炎症性肺损伤， 发病率和死亡率高得令人无法接受的疾病。我们的支持性数据表明， NFATc 3的消融和药物抑制有利于增强肺屏障功能， 减少细胞因子释放和嗜酸性炎症，改善动脉氧合， ARDS/ALI小鼠模型。基于这些和其他临床相关的支持性数据，我们认为， 巨噬细胞中的NFATc 3活化诱导介导启动的炎性基因表达， ALI/ARDS的强度和持续时间。我们提出了三个具体目标来解决这个假设： 具体目的1：确定NFATc 3缺失保护免受ALI/ARDS的机制。 我们将采用遗传上确定的小鼠，其总体上缺乏NFATc 3，并进行明确的小鼠模型， ALI/ARDS。我们还将利用已建立的NFATc 3 KO/WT骨髓嵌合体小鼠的方法， 巨噬细胞被动过继转移肺巨噬细胞，我们将构建新的条件 巨噬细胞选择性NFATc 3-/-小鼠，以确定巨噬细胞NFATc 3缺乏对 肺水肿的产生、动脉血氧饱和度和死亡率。 具体目标2：确定巨噬细胞NFATc 3激活有助于 发生渗透性肺水肿。为了确定巨噬细胞如何参与 在ALI/ARDS的肺水肿和严重低氧血症中，我们建议检查NFATc 3的作用 调节巨噬细胞表达肿瘤坏死因子相关凋亡诱导配体（TRAIL），其抑制 肺泡液清除率（AFC），以及肺巨噬细胞产生的介质破坏屏障 肺微血管内皮功能。 具体目的3：确定新型细胞渗透性肽NFAT抑制剂在保护细胞凋亡中的功效。 针对小鼠模型中的ALI/ARDS样病理生理学。我们的团队开发了一种新型的NFAT 抑制剂CP 9-ZIZIT，其具有令人印象深刻的低Kd值2.6 nM，更好的细胞渗透性，更高的结合 亲和性，以及比其他可用试剂改善的抗蛋白水解稳定性。我们希望能证明 单独使用CP 9-ZIZIT或与PARP 1抑制剂组合使用CP 9-ZIZIT的治疗将减弱肺动脉高压。 炎症和肺泡毛细血管屏障破坏。 我们希望这项研究将提供有价值的临床前数据，阻断NFATc 3激活， 巨噬细胞是预防高危患者群体中的ARDS的有效治疗剂 改善中重度ARDS/ALI患者的临床预后。