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，NFATc3在巨噬细胞中的遗传消融和药物抑制是 有利于维持肺泡-毛细血管屏障功能，防止炎性细胞因子释放和 内毒素和盲肠结扎穿刺术中中性粒细胞炎症、动脉氧合改善和存活率 急性呼吸窘迫综合征小鼠模型。在这里，我们建议确定下游分子靶标的颗粒细节 使用两次打击的小鼠模型、人肺巨噬细胞和ARDS患者的BALF对NFATc3进行研究。我们队 已经开发出一种新型的无毒细胞通透性钙调神经磷酸酶抑制肽(CNI)(CNI103)，它可以阻断 激活巨噬细胞中的NFATc3并减轻小鼠的ARDS。我们建议确定分子 临床前二次击打小鼠模型、人肺巨噬细胞和患者BALF的NFATc3靶点 达到柏林的ARDS标准。我们的中心假设是钙调神经磷酸酶依赖的NFAT的激活 在巨噬细胞中调节ALI/ARDS的发生，并抑制NFAT的激活 多肽钙调神经磷酸酶抑制剂(CNI)可以减轻疾病的严重程度。我们提出两个具体目标： 特异性目标1：确定NFATc3-钙调神经磷酸酶激活途径的下游分子靶点 ALI/ARDS时肺巨噬细胞的变化新的初步数据显示，细胞外脂含量 BALF中的小泡(EV)依赖NFATc3并介导肺微血管屏障功能的破坏 内皮细胞(MVEC)。在SA1中，我们将1)确定NFAT调节的血脂介体和 参与脂类代谢的酶，2)决定这些介质是否包装在细胞外 小泡，3)评估EVS是否介导了导致通透性的细胞间通讯 肺水肿和4)确定阻断NFAT激活是否可以防止肺损伤和炎症 临床相关的ARDS小鼠和细胞模型。 具体目标2：确定一种优化的细胞通透性钙调神经磷酸酶抑制剂的有效性和安全性， 在临床相关的感染性和非感染性小鼠模型中，该药物可防止NFAT激活 ALI/ARDS。我们将测试细胞通透性钙调神经磷酸酶多肽抑制剂是否能预防和逆转肺损伤。 以及ARDS小鼠模型中的炎症。我们还将评估药物动力学和药效学。 (PK/PD)特性、细胞选择性、安全性、有效性和在预防和逆转肺损伤中的作用 临床前ARDS小鼠模型。 这些研究将促进对NFATc3激活在巨噬细胞中的重要作用的了解 等肺细胞类型在ALI/ARDS发病机制中的作用。我们期待着从这些知识中获得 这些研究将建立NFATc3作为调节EV介导的细胞-细胞的新的治疗靶点 通过控制具有生物活性的脂类和蛋白质介体的组成来实现相互作用。