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

• 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
• 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

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 高死亡率率，没有有益的靶向疗法。对于退伍军人和当前来说，这是一个紧迫的问题 估计表明，每年的退伍军人将死于ARDS，而不是死于肺癌。尽管 这是没有用于ARDS的药理学疗法。需要对发病机理的新见解 加深我们对导致ARD和发展新颖的基本机制的理解 治疗。 ARD的主要病理特征是激活组织因子（TF）途径 空域中的凝结。 TF是一种整体膜蛋白，均启动外部凝血 级联并提供多种非致凝功能，包括促进细胞粘合剂和通过 与整合蛋白蛋白的相互作用。几十年来，包括TF途径在内的Procagulant途径一直是 作为ARDS损伤机制实施。但是，许多抗凝剂的临床踪迹尚未 在这种方法中显示出好处。一个潜在的解释是，空域中的TF受到ARD的保护。 我们的初步体内和体外数据表明，肺上皮的丧失，但不是巨噬细胞，TF会导致损失 上皮屏障完整性，细胞表面β1整合素和异常细胞粘合剂的降低。反过来， TF在肺泡上皮II型细胞中的过表达恢复了屏障完整性。另外，用 重组因子VII（RFVII），主要的TF配体进入空域，使小鼠中的渗透性减弱 ARDS的模型。这些观察结果共同改变了理解TF的作用的重大转变 ARDS中的途径，几乎普遍认为有害的途径。该提议的目标 正在测试（1）肺上皮TF丧失的新假设导致上皮屏障完整性的丧失 通过破坏整联蛋白蛋白和（2）在空域中的TF上调维持上皮屏障 直接肺损伤模型中的完整性。因此，将RFVII当地交付到领空可以代表独特的 预防和治疗ARDS的治疗方法。使用牙槽的新型转基因小鼠 上皮II型细胞特异性缺失或TF的过表达我们将定义体内细胞特异性机制 TF对肺上皮屏障完整性的影响。我们还将使用新型的TF敲除肺上皮细胞系 研究TF和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 通过表达新型TF突变体库来完整性。最后，我们将使用与临床相关的人类模型 ARDS（离体灌注人肺）测试RFVII的治疗潜力，以拧紧上皮屏障 人类ards。我们的实验设计既是机械的又是翻译，并且利用了新型转基因 小鼠模型，细胞系以及人类肺损伤模型，既了解TF效应的机制 肺以及测试RFVII的治疗潜力。该建议利用了独特的技能 范德比尔特（Vanderbilt）可用的资源，包括最先进的计算建模和广告 复杂的动力学测量，从而确保了这些研究的结果将推进现场，并且 为为有ARDS的退伍军人开发有针对性的治疗提供必要的基础。

项目成果

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

• DOI: 10.1107/s1600536810030473
• 发表时间: 2010-08-18
• 期刊: Acta crystallographica. Section E, Structure reports online
• 作者: Bruni B;Bartolucci G;Ciattini S;Coran S
• 通讯作者: Coran S

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: 10.1152/ajplung.00239.2021
• 发表时间: 2021
• 期刊: American journal of physiology. Lung cellular and molecular physiology
• 作者: Shimoda,LarissaA;Bai,Chunxue;Bartlett,NathanW;Bastarache,JulieA;Feghali-Bostwick,Carol;Kuebler,WolfgangM;Prakash,YS;Schmidt,EricP;Morty,RoryE
• 通讯作者: Morty,RoryE

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

• DOI: 10.14814/phy2.14589
• 发表时间: 2020-11
• 期刊: Physiological reports
• 影响因子: 2.5
• 作者: Shaver CM;Landstreet SR;Pugazenthi S;Scott F;Putz N;Ware LB;Bastarache JA
• 通讯作者: Bastarache JA