Potential Protective Mechanisms of Tissue Factor in Acute Lung Injury

组织因子在急性肺损伤中的潜在保护机制

基本信息

批准号：
10045936
负责人：
Julie Anne Bastarache
金额：
--
依托单位：
VETERANS HEALTH ADMINISTRATION
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-10-01 至 2021-09-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10045936
关键词：
Abnormal Cell Actins Acute Lung Injury Acute respiratory failure Adhesions Adult Respiratory Distress Syndrome Affect Alveolar Alveolar Macrophages American Animals Anticoagulants Attenuated Binding Biological Models Blood Coagulation Factor VII Blood capillaries Cancer Biology Cell Adhesion Cell Line Cell Migration Pathway Cell surface Cells Clinical Clinical Research Coagulants Coagulation Process Complex Computer Models Cultured Cells Data Development Ensure Epithelial Epithelial Cells Experimental Designs Extracellular Domain Fibrin Foundations Generations Genetic Goals Human In Vitro Inflammatory Injury Integral Membrane Protein Integrin Binding Integrins Intravenous Klebsiella pneumoniae Knock-out Knowledge Laboratories Lead Libraries Life Ligand Binding Ligands Link Lipopolysaccharides Lung Malignant neoplasm of lung Measurement Mediating Methods Modeling Molecular Mus Pathogenesis Pathologic Pathway interactions Patients Permeability Pharmacologic Substance Pharmacology Prevention approach Proteins Publishing Recombinants Research Personnel Resources Role Structural Protein Testing Therapeutic Thrombin Thromboplastin Transgenic Mice Type II Epithelial Receptor Cell Up-Regulation Veterans Work alveolar epithelium cell injury cell motility clinical efficacy clinically relevant experimental study extracellular human model in vivo insight lung injury mortality mouse model mutant novel novel therapeutic intervention novel therapeutics overexpression protective effect response skills targeted treatment therapeutic evaluation

项目摘要

Acute Respiratory Distress Syndrome (ARDS) is a common cause of acute respiratory failure that carries a high mortality rate and has no beneficial targeted therapies. It is a pressing problem for Veterans and current estimates suggest that more Veterans will die from ARDS each year than will die from lung cancer. Despite this, there are no pharmacologic therapies for ARDS. New insights into the pathogenesis are needed to deepen our understanding of the underlying mechanisms that lead to ARDS as well as to develop novel therapeutics. A major pathologic feature of ARDS is activation of the Tissue Factor (TF) pathway of coagulation in the airspace. TF is an integral membrane protein that both initiates the extrinsic coagulation cascade and serves several non-coagulant functions including promoting cell adhesion and migration through interactions with integrin proteins. For decades, procoagulant pathways, including the TF pathway, have been implicated as a mechanism of injury in ARDS. However, a number of clinical trails of anti-coagulants have not shown a benefit with this approach. One potential explanation is that TF in the airspace is protective in ARDS. Our preliminary in vivo and in vitro data show that loss of lung epithelial, but not macrophage, TF causes loss of epithelial barrier integrity, decreased cell surface β1 integrin and abnormal cell adhesion. Conversely, overexpression of TF in alveolar epithelial type II cells restores barrier integrity. In addition, treatment with recombinant factor VII (rFVII), the primary TF ligand, into the airspace attenuates permeability in a mouse model of ARDS. Together, these observations represent a major shift in understanding the role of the TF pathway in ARDS, a pathway that has almost universally been considered harmful. The goals of this proposal are to test the novel hypotheses that (1) loss of lung epithelial TF leads to loss of epithelial barrier integrity through disruption of integrin proteins and (2) upregulation of TF in the airspace maintain epithelial barrier integrity in a direct lung injury model. Therefore, local delivery of rFVII to the airspace could represent a unique therapeutic approach for the prevention and treatment of ARDS. Using novel transgenic mice with alveolar epithelial type II cell specific deletion or overexpression of TF we will define the in vivo cell specific mechanism of TF effects on lung epithelial barrier integrity. We will also use a novel TF knockout lung epithelial cell line to study the molecular interactions between TF and β1 integrin and the role of TF in regulating epithelial barrier integrity by expressing a library of novel TF mutants. Finally we will use a clinically relevant human model of ARDS (ex vivo perfused human lung) to test the therapeutic potential of rFVII to tighten the epithelial barrier in human ARDS. Our experimental design is both mechanistic and translational and utilizes novel transgenic mouse models, cell lines as well a human lung injury model to both understand the mechanism of TF effects in the lung as well as to test the therapeutic potential of rFVII. This proposal takes advantage of the unique skills and resources available at Vanderbilt including state of the art computational modeling as well as adhesion complex dynamics measurements thus ensuring that results from these studies will advance the field and provide the necessary foundation for development of targeted therapeutics for Veterans with ARDS.

急性呼吸窘迫综合征 (ARDS) 是急性呼吸衰竭的常见原因，可导致死亡率高，并且没有有益的靶向治疗，这对退伍军人和当前来说都是一个紧迫的问题。据估计，每年死于急性呼吸窘迫综合征的退伍军人比死于肺癌的退伍军人还要多。因此，目前还没有针对 ARDS 的药物疗法。加深我们对导致 ARDS 的潜在机制的理解并开发新的 ARDS 的一个主要病理特征是组织因子 (TF) 途径的激活。 TF 是一种完整的膜蛋白，可启动外源性凝血。级联并提供多种非凝血功能，包括通过促进细胞粘附和迁移几十年来，促凝血途径（包括 TF 途径）一直在研究。然而，许多抗凝剂的临床试验尚未发现它是 ARDS 的损伤机制。一种可能的解释是，空域中的 TF 对 ARDS 具有保护作用。我们的初步体内和体外数据表明，肺上皮细胞（而非巨噬细胞）的损失会导致 TF 损失上皮屏障完整性、细胞表面 β1 整合素减少和细胞粘附异常。 TF 在 II 型肺泡上皮细胞中的过度表达可恢复屏障完整性。重组因子 VII (rFVII)（主要 TF 配体）进入空气空间会减弱小鼠的通透性这些观察结果共同代表了理解 TF 作用的重大转变。 ARDS 中的途径，几乎普遍被认为是有害的途径。目的是测试以下新假设：(1) 肺上皮 TF 的丧失导致上皮屏障完整性的丧失通过破坏整合素蛋白和 (2) 上调空腔中的 TF 维持上皮屏障因此，将 rFVII 局部递送到空域可能代表了一种独特的方法。使用具有肺泡的新型转基因小鼠预防和治疗 ARDS 的治疗方法。上皮II型细胞特异性缺失或过度表达TF，我们将定义体内细胞特异性机制我们还将使用新型 TF 敲除肺上皮细胞系来研究 TF 对肺上皮屏障完整性的影响。研究TF与β1整合素之间的分子相互作用以及TF在调节上皮屏障中的作用最后，我们将使用临床相关的人类模型来表达新的 TF 突变体库。 ARDS（离体灌注人肺）测试 rFVII 收紧上皮屏障的治疗潜力我们的实验设计既是机械性的又是转化性的，并利用了新颖的转基因技术。小鼠模型、细胞系以及人类肺损伤模型，以了解 TF 作用的机制该提案利用了独特的技术。范德比尔特大学提供的资源包括最先进的计算模型以及粘附力复杂的动力学测量，从而确保这些研究的结果将推动该领域的发展为患有 ARDS 的退伍军人开发靶向治疗方法提供必要的基础。

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

The NLRP3 inflammasome in macrophages is stimulated by cell-free hemoglobin.

巨噬细胞中的 NLRP3 炎症小体受到游离血红蛋白的刺激。

DOI：
发表时间：
2020
期刊：
影响因子：
2.5
作者：
Shaver, Ciara M;Landstreet, Stuart R;Pugazenthi, Sangamithra;Scott, Fiona;Putz, Nathan;Ware, Lorraine B;Bastarache, Julie A
通讯作者：
Bastarache, Julie A

Announcing the Editorial Board Fellowship Program of the American Journal of Physiology-Lung Cellular and Molecular Physiology.

宣布《美国生理学杂志-肺细胞和分子生理学》编委会奖学金计划。

DOI：
发表时间：
2021
期刊：
影响因子：
0
作者：
Shimoda, Larissa A;Bai, Chunxue;Bartlett, Nathan W;Bastarache, Julie A;Feghali;Kuebler, Wolfgang M;Prakash, Y S;Schmidt, Eric P;Morty, Rory E
通讯作者：
Morty, Rory E

2-(1,2,3,4-Tetra-hydro-1-naphth-yl)imidazolium chloride monohydrate.

2-(1,2,3,4-四氢-1-萘基)氯化咪唑鎓一水合物。