Proteolysis in the Pathogenesis of ARDS

ARDS 发病机制中的蛋白水解作用

基本信息

批准号：
10543482
负责人：
Gregory Paul Downey
金额：
$ 69.61万
依托单位：
NATIONAL JEWISH HEALTH
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-12-22 至 2025-11-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10543482
关键词：
Acceleration Acids Acute Acute Lung Injury Acute Respiratory Distress Syndrome Address Adhesions Affect Affinity Alveolar Animal Model Area Basal Cell Basement membrane Binding Biological Markers Blood Blood capillaries Cell Culture Techniques Cell secretion Cells Characteristics Clinical Clinical Data Clinical Research Clinical/Radiologic Complement Data Directed Molecular Evolution Disease Disease Progression Distal Epithelial Cells Epithelium Event Extracellular Matrix Failure Fibroblasts Gases Health Care Costs Healthcare Human Immunohistochemistry Impairment In Vitro Inflammation Injury Intercellular Junctions Interdisciplinary Study Kinetics Knowledge Laboratories Life Link Liquid substance Lung Lung diseases MMP3 gene Macrophage Matrix Metalloproteinase Inhibitor Matrix Metalloproteinases Mediating Membrane Mus Organ Donor Pathogenesis Pathology Pathology processes Pathway interactions Patients Peptide Hydrolases Permeability Persons Physiological Play Population Pre-Clinical Model Process Production Proteolysis Publishing Pulmonary Edema Recovery Research Research Proposals Resistance Resolution Role Sampling Slice Stimulus Stromelysin 1 Structure of parenchyma of lung Supportive care Testing Therapeutic Therapeutic Agents Therapeutic Intervention Time Tissue Inhibitor of Metalloproteinase-1 Tissue Inhibitor of Metalloproteinases Tissues Transgenic Mice United States National Institutes of Health Variant Work alveolar epithelium aspirate biobank care burden clinically relevant conditional knockout design epithelial injury epithelial repair experimental study high risk human disease improved outcome in vivo Model indexing influenza infection influenzavirus inhibitor inhibitor therapy injury and repair lung injury lung repair mortality mouse model neutrophil novel novel therapeutic intervention overexpression patient subsets personalized strategies pleiotropism pre-clinical predictive marker preservation prevent repaired response restoration single cell analysis single-cell RNA sequencing specific biomarkers therapeutic evaluation transcriptomics translational applications ventilation

项目摘要

PROJECT SUMMARY/ABSTRACT The Acute respiratory distress syndrome (ARDS) is the severest form of acute lung injury (ALI), and is characterized by injury to the alveolar-capillary unit and compromised alveolar epithelial integrity, leading to high permeability pulmonary edema and neutrophilic alveolar inflammation. Failure to repair the damaged alveolar membrane leads to significant mortality. Studies from several laboratories, including our own, have found that induction of ALI in animal models is associated with increased expression of matrix metalloproteinase-3 (MMP-3), a protease that has been shown to directly target cell-cell junctions and to degrade basement membranes, although it was unknown at what stages of ALI pathology this process was most critical. Other laboratories and ours have also found that transgenic mice lacking MMP-3 are resistant to lung injury induced by multiple stimuli, indicating that inhibitors of MMP-3 have potential as therapeutic agents for ARDS, although such a therapeutic strategy would need to be highly selective, as other MMPs have been shown to play key roles in repair of lung injury. In this translational application, we propose experiments which will bridge these critical gaps. In Aim 1, we will use physiologically relevant cell culture and animal models of ALI/ARDS including novel transgenic mice to define how MMP-3 affects the resolution of epithelial injury in response to acid aspiration or influenza infection. In Aim 2, we will use our directed molecular evolution platform to create variants of Tissue Inhibitor of Metalloproteinase-1 (TIMP-1) with greatly increased affinity for MMP-3 and decreased binding to beneficial MMPs, and we will define the therapeutic utility of these TIMP variants in mouse models of ALI/ARDS. In Aim 3, we will interrogate clinical samples of ARDS to determine the precise stages of human disease progression at which therapeutic intervention would be most beneficial. Our research proposal will define a newly-discovered mechanism by which MMP-3 drives ARDS pathology through an integrated research plan that links single cell RNA sequencing of cell populations isolated from ARDS patient tissue, cell culture and animal models of ARDS, and analysis of relevant ARDS biomarkers and ex vivo culture of precision cut human lung slices, and will develop a novel selective inhibitor with immediate therapeutic potential. Our multidisciplinary research team is uniquely suited to address these questions. Our work will address critical mechanisms underlying ALI/ARDS, will use this knowledge to develop personalized strategies that will identify patients at highest risk of developing ALI/ARDS, and will create therapeutic approaches based on the pivotal role of MMP-3 in the pathogenesis of epithelial injury leading to ALI/ARDS.

项目摘要/摘要急性呼吸窘迫综合征（ARDS）是急性肺损伤（ALI）的最严重形式，是以肺泡毛细血管单元的损伤为特征，并损害了肺泡上皮完整性，导致高渗透性肺水肿和嗜中性肺泡炎症。无法修复受损的牙槽膜导致显着死亡。来自包括我们自己在内的几个实验室的研究发现动物模型中ALI的诱导与基质的表达增加有关金属蛋白酶-3（MMP-3），一种蛋白酶已被证明直接靶向细胞 - 细胞连接和至降解地下室膜，尽管在Ali病理的哪个阶段尚不清楚此过程是最关键的。其他实验室和我们的人也发现，缺乏MMP-3的转基因小鼠对多种刺激引起的肺损伤，表明MMP-3的抑制剂具有治疗剂的潜力对于ARDS，尽管这种治疗策略需要高度选择性，就像其他MMP一样显示在修复肺损伤中起关键作用。在此翻译应用中，我们提出了实验将弥合这些关键的差距。在AIM 1中，我们将使用与生理相关的细胞培养和动物模型 Ali/Ards包括新型转基因小鼠，以定义MMP-3如何影响上皮损伤的分辨率对酸抽吸或流感感染的反应。在AIM 2中，我们将使用定向的分子进化建立金属蛋白酶-1（TIMP-1）的组织抑制剂的平台，对 MMP-3并减少与有益MMP的结合，我们将定义这些TIMP的治疗效用 ALI/ARD的小鼠模型中的变体。在AIM 3中，我们将询问ARDS的临床样本以确定治疗干预最有益的人类疾病进展的精确阶段。我们的研究建议将定义一种新发现的机制一个综合研究计划，该计划将从ARDS分离的细胞群的单细胞RNA测序联系起来 ARDS的患者组织，细胞培养和动物模型，并分析相关的ARDS生物标志物和离体精确的培养人类肺切片，并将形成一种新颖的选择性抑制剂治疗潜力。我们的多学科研究团队非常适合解决这些问题。我们的工作将解决ALI/ARDS的关键机制，将使用此知识来发展个性化策略将确定患者患ALI/ARD的风险最高的患者，并将创建治疗性基于MMP-3在导致ALI/ARD的上皮损伤发病机理中的关键作用。