REGULATION OF THE MACROPHAGE INFLAMMATORY PHENOTYPE IN ARDS

ARDS 中巨噬细胞炎症表型的调节

• 批准号: 10455872
• 负责人: John W Christman
• 金额: $ 56.88万
• 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 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10455872
• 关键词: Ablation; Acute Respiratory Distress Syndrome; Address; Affect; Alveolar; Alveolar Macrophages; Amino Acids; Arachidonic Acids; Bacteremia; Berlin; Blood capillaries; COVID-19; Calcineurin; Calcineurin inhibitor; Capillary Permeability; Cell Communication; Cell model; Cells; Complication; Critical Illness; Data; Development; Disease; Endocytosis; Endothelium; Enzymes; Epithelial Cells; Escherichia coli; Functional disorder; Genetic; Goals; Health; Human; Inflammation; Inflammation Mediators; Inflammatory; Injury; Intensive Care Units; Interleukin-6; Intravenous; Knowledge; Lead; Life; Lipids; Lipopolysaccharides; Liquid substance; Lung; Lung Lavage Fluid; Measures; Mediating; Mediator of activation protein; Metabolism; Modeling; Molecular; Molecular Target; Mus; Outcome; PTGS2 gene; Pathogenesis; Pathway interactions; Patients; Peptides; Permeability; Pharmacological Treatment; Pharmacology; Phenotype; Prevention approach; Property; Proteins; Proteolysis; Public Health; Publishing; Pulmonary Edema; Pulmonary Inflammation; Research; Resistance; Resolution; Role; Route; Safety; Severity of illness; Staphylococcus aureus; Structure; Supportive care; T-Cell Activation; TNF gene; Testing; Transcriptional Activation; Ventilator-induced lung injury; Work; alveolar epithelium; base; cecal ligation puncture; cell type; cellular targeting; clinically relevant; cytokine; cytokine release syndrome; efficacy testing; extracellular vesicles; improved; inhibitor; lipid mediator; lipid metabolism; lung injury; lung microvascular endothelial cells; macrophage; meetings; mortality; mouse model; neutrophil; new therapeutic target; novel; novel strategies; nuclear factors of activated T-cells; pandemic disease; pharmacokinetics and pharmacodynamics; pre-clinical; prevent; transcription factor NF-AT c3; treatment response; ventilation

Our published data show that genetic ablation and pharmacologic inhibition of NFATc3 in macrophages is beneficial in maintaining alveolar-capillary barrier function, prevents inflammatory cytokine release and neutrophilic inflammation, improved arterial oxygenation and survival in the LPS and cecal ligation puncture mouse models of ARDS. Here, we propose to determine the granular details of the downstream molecular targets of NFATc3 using a 2-hit mouse model, human lung macrophages, and BALF from patients with ARDS. Our team has developed a novel non-toxic cell permeable calcineurin inhibitory (CNI) peptide (CNI103) that blocks activation of NFATc3 in macrophages and mitigates ARDS in mice. We propose to determine the molecular targets of NFATc3 using a pre-clinical 2-hit mouse model, human lung macrophages, and BALF from patients meeting the Berlin criterion for ARDS. Our central hypothesis is that activation of calcineurin-dependent NFAT in macrophages regulates the development of ALI/ARDS, and inhibition of NFAT activation by a novel peptide calcineurin inhibitor (CNI) lessens disease severity. We propose two specific aims: Specific Aim 1: To delineate the downstream molecular targets of NFATc3-Calcineurin activation pathway in pulmonary macrophages during ALI/ARDS. Novel preliminary data show that the lipid content of extracellular vesicles (EVs) in BALF are NFATc3 dependent and mediate disruption of barrier function in lung microvascular endothelial cells (MVEC). In SA1, we will 1) determine the spectrum of NFAT regulated lipids mediators and enzymes involved in lipid metabolism, 2) determine whether these mediators are packaged in extracellular vesicles, 3) assess whether EVs mediate the the cell-to-cell communication that results in permeability pulmonary edema and 4) determine whether blocking NFAT activation prevents lung injury and inflammation in clinically relevant mouse and cellular models of ARDS. Specific Aim 2: To determine the efficacy and safety of an optimized cell permeable calcineurin inhibitor, which prevents NFAT activation, in clinically relevant infectious and non-infectious mouse models of ALI/ARDS. We will test whether cell permeable calcineurin peptide inhibitors prevent and reverse lung injury and inflammation in mouse models of ARDS. We will also assess the pharmacokinetic and pharmacodynamics (PK/PD) properties, cellular selectivity, safety, efficacy, and potency in preventing and reversing lung injury in preclinical mouse models of ARDS. These studies will advance knowledge about the essential role of NFATc3 activation in macrophages and other lung cell types in the pathogenesis of ALI/ARDS. We anticipate that knowledge gained from these studies will establish NFATc3 as a novel therapeutic target that regulates EV-mediated cell-cell interactions by governing the composition of biologically active lipid and protein mediators.

我们发表的数据表明，巨噬细胞中NFATC3的遗传消融和药理抑制是 有益于维持肺泡毛细血管屏障功能，可防止炎症细胞因子释放和 中性粒细胞炎症，改善的动脉氧合和LPS和Cecal结扎穿刺的生存 ARDS的鼠标模型。在这里，我们建议确定下游分子靶标的颗粒细节 使用2击小鼠模型，人类肺巨噬细胞和来自ARDS患者的BALF的NFATC3。我们的团队 已经开发了一种新型的非毒性细胞渗透性钙化蛋白抑制（CNI）肽（CNI103），该肽阻止了 NFATC3在巨噬细胞中的激活和减轻小鼠的ARDS的激活。我们建议确定分子 NFATC3的靶标使用临床前的2击小鼠模型，人肺巨噬细胞和患者的BALF的靶标 满足柏林ARDS标准。我们的中心假设是钙调蛋白依赖性NFAT的激活 在巨噬细胞中，调节ALI/ARD的发展，并通过新颖的抑制NFAT激活 肽钙调蛋白抑制剂（CNI）减轻了疾病的严重程度。我们提出了两个具体目标： 特定目的1：描绘NFATC3-钙调蛋白激活途径的下游分子靶标 ALI/ARDS期间的肺巨噬细胞。新型初步数据表明细胞外脂质含量 BALF中的囊泡（EV）是NFATC3依赖性的，并介导了肺微血管中屏障功能的破坏 内皮细胞（MVEC）。在SA1中，我们将1）确定NFAT调节的脂质介质和 参与脂质代谢的酶，2）确定这些介体是否包装在细胞外 囊泡，3）评估EVS是否介导导致渗透性的细胞间通信 肺水肿和4）确定阻断NFAT激活是否阻止了肺损伤和炎症 临床相关的小鼠和ARDS的细胞模型。 特定目的2：确定优化细胞可渗透性钙调神经抑制剂的功效和安全性， 它可以防止NFAT激活，在临床相关的传染和非感染小鼠模型中 Ali/ards。我们将测试可渗透性钙调神经蛋白肽抑制剂预防和反向肺损伤 和ARDS小鼠模型中的炎症。我们还将评估药代动力学和药效学 （PK/PD）特性，细胞选择性，安全性，有效性以及预防和逆转肺损伤的效力 ARDS的临床前小鼠模型。 这些研究将提高有关NFATC3激活在巨噬细胞中的基本作用的知识 ALI/ARD的发病机理中的其他肺细胞类型。我们预计从中获得的知识 这些研究将建立NFATC3作为调节EV介导的细胞细胞的新型治疗靶标 通过管理生物活性脂质和蛋白质介质的组成来进行相互作用。