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.

项目概要/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，其特点是肺泡毛细血管单位损伤和肺泡上皮完整性受损，导致高渗透性肺水肿和中性粒细胞性肺泡炎症。未能修复损坏的肺泡膜导致显着的死亡率。来自多个实验室（包括我们自己的实验室）的研究表明发现动物模型中 ALI 的诱导与基质表达增加有关金属蛋白酶-3 (MMP-3)，一种蛋白酶，已被证明可以直接靶向细胞与细胞的连接并降解基底膜，尽管尚不清楚该过程发生在 ALI 病理学的哪个阶段最关键的。其他实验室和我们的实验室也发现缺乏 MMP-3 的转基因小鼠对多重刺激引起的肺损伤，表明 MMP-3 抑制剂具有作为治疗药物的潜力对于 ARDS，尽管这种治疗策略需要具有高度选择性，正如其他 MMP 一样研究显示其在肺损伤修复中发挥关键作用。在这个转化应用中，我们提出了实验将弥合这些关键差距。在目标 1 中，我们将使用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消退对酸吸入或流感感染的反应。在目标 2 中，我们将使用定向分子进化平台创建金属蛋白酶组织抑制剂 1 (TIMP-1) 的变体，其亲和力大大增加 MMP-3 和与有益 MMP 的结合减少，我们将定义这些 TIMP 的治疗效用 ALI/ARDS 小鼠模型的变体。在目标 3 中，我们将询问 ARDS 的临床样本以确定人类疾病进展的精确阶段，在该阶段治疗干预最为有益。我们的研究提案将定义一种新发现的机制，MMP-3 通过该机制驱动 ARDS 病理学一项综合研究计划，将对从 ARDS 中分离出的细胞群进行单细胞 RNA 测序 ARDS 患者组织、细胞培养物和动物模型，以及相关 ARDS 生物标志物和离体分析精密切割人肺切片的培养，并将开发一种新型选择性抑制剂，具有立即治疗潜力。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，将利用这些知识来开发个性化的确定发生 ALI/ARDS 风险最高的患者的策略，并创造治疗方法该方法基于 MMP-3 在导致 ALI/ARDS 的上皮损伤发病机制中的关键作用。