REGULATION OF THE MACROPHAGE INFLAMMATORY PHENOTYPE IN ARDS

ARDS 中巨噬细胞炎症表型的调节

基本信息

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
项目类别：
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

项目摘要

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 的基因消除和药理学抑制是有益于维持肺泡毛细血管屏障功能，防止炎症细胞因子释放和中性粒细胞炎症，改善脂多糖和盲肠结扎穿刺中的动脉氧合和存活率 ARDS 小鼠模型。在这里，我们建议确定下游分子靶标的粒度细节使用 2 次命中小鼠模型、人肺巨噬细胞和来自 ARDS 患者的 BALF 来检测 NFATc3。我们的团队开发了一种新型无毒细胞渗透性钙调神经磷酸酶抑制 (CNI) 肽 (CNI103)，可阻断激活巨噬细胞中的 NFATc3 并减轻小鼠的 ARDS。我们建议确定分子使用临床前 2 次命中小鼠模型、人肺巨噬细胞和来自患者的 BALF 来确定 NFATc3 的靶标符合 ARDS 柏林标准。我们的中心假设是钙调神经磷酸酶依赖性 NFAT 的激活巨噬细胞中的一种新型药物可调节 ALI/ARDS 的发展，并抑制 NFAT 激活肽钙调神经磷酸酶抑制剂（CNI）可减轻疾病的严重程度。我们提出两个具体目标：具体目标 1：描绘 NFATc3-钙调神经磷酸酶激活途径的下游分子靶标 ALI/ARDS 期间的肺巨噬细胞。新的初步数据表明，细胞外的脂质含量 BALF 中的囊泡 (EV) 依赖于 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 调节的脂质介质的谱和参与脂质代谢的酶，2）确定这些介质是否包装在细胞外囊泡，3) 评估 EV 是否介导导致渗透性的细胞间通讯肺水肿和 4) 确定阻断 NFAT 激活是否可以预防肺损伤和炎症 ARDS 临床相关小鼠和细胞模型。具体目标 2：确定优化的细胞渗透性钙调神经磷酸酶抑制剂的功效和安全性，在临床相关的感染性和非感染性小鼠模型中，可防止 NFAT 激活急性肺损伤/急性呼吸窘迫综合征。我们将测试细胞渗透性钙调磷酸酶肽抑制剂是否可以预防和逆转肺损伤和 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、有效性和预防和逆转肺损伤的效力 ARDS 临床前小鼠模型。这些研究将增进人们对 NFATc3 激活在巨噬细胞中的重要作用的认识和其他肺细胞类型在 ALI/ARDS 发病机制中的作用。我们预计从以下方面获得的知识这些研究将确立 NFATc3 作为调节 EV 介导的细胞的新型治疗靶点通过控制生物活性脂质和蛋白质介质的组成来相互作用。