Endothelial thrombospondin-1 in matrix proteolysis during Pseudomonal lung injury

内皮血小板反应蛋白-1 在假单胞菌肺损伤期间基质蛋白水解中的作用

• 批准号: 10461702
• 负责人: Jill Ann Zupetic
• 金额: $ 1.63万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2021-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10461702
• 关键词: Accounting; Acute; Adherens Junction; Admission activity; Adult Respiratory Distress Syndrome; Advanced Development; Alveolar; Animal Model; Bacterial Infections; Bacterial Pneumonia; Biology; Blood Platelets; Blood Vessels; Blood capillaries; Cell Line; Cells; Clinical; Complex; Critical Illness; DNA Damage; Data; Diffuse; Dipeptides; Dose; Elastases; Endothelial Cells; Endothelium; Environment; Epithelial Cells; Exhibits; Exopeptidase; Extracellular Matrix; Functional disorder; Future; Glycoproteins; Human; Hypoxemia; In Vitro; Infection; Inflammation; Inflammatory; Injury; Intercellular Junctions; Kinetics; Knockout Mice; Knowledge; Laboratories; Lead; Leukocyte Elastase; Literature; Lower Respiratory Tract Infection; Lung; Mentors; Metalloproteases; Modeling; Morbidity - disease rate; Mus; Myeloid Cells; Nosocomial Infections; Outcome; Parents; Patients; Peptide Hydrolases; Physicians; Pneumonia; Proteins; Proteolysis; Pseudomonas aeruginosa; Pseudomonas aeruginosa infection; Pseudomonas aeruginosa pneumonia; Reagent; Recovery; Refractory; Regulation; Research; Respiratory Failure; Risk Factors; Role; Scientist; Serine Protease; Small Interfering RNA; Syndrome; Testing; Thrombospondin 1; Tight Junctions; Tissue imaging; Tissues; Training; Type II Secretion System Pathway; Universities; Vascular Permeabilities; Virulence Factors; acute hypoxemic respiratory failure; base; cadherin 5; cell injury; cell type; cellular imaging; combat; improved; in vivo; in vivo Model; inhibitor/antagonist; knock-down; lung injury; microbial; mortality; mouse genetics; mutant; neutrophil; novel; occludin; pathogen; respiratory; response; response to injury; severe injury; skills; therapeutic target; ventilator-associated pneumonia

Project Summary/Abstract: Acute Respiratory Distress Syndrome (ARDS) is a syndrome characterized by rapidly progressive respiratory failure and refractory hypoxemia and is a major cause of morbidity and mortality in critically ill patients worldwide. The most common risk factor for ARDS is severe bacterial pneumonia and patients with ARDS are also frequently plagued by nosocomial infections including lower respiratory tract infections (LRTIs). Pseudomonas aeruginosa (PA) is a common ICU pathogen and PA pneumonia is associated with increased mortality in ventilator associated pneumonia compared to other gram-negative pathogens. PA has been shown to cause extensive proteolytic injury leading to tissue damage and degradation of the alveolar-capillary barrier in animal models, in part through secreted exoproteases such as LasB. The metalloprotease LasB has also been shown to degrade junctional proteins important in barrier integrity. While pathogen derived virulence factors have been extensively characterized, there is limited knowledge regarding the host mechanisms that protect against proteolytic injury and vascular permeability in the lungs. Thrombospondin-1 (TSP-1) is a matrix glycoprotein with a variety of functions including regulation of inflammation. TSP-1 has been previously shown to inhibit host- derived serine proteases and the pathogen-derived exoprotease, LasB. TSP-1 is expressed by lung endothelial cells but its role in pathogen-induced injury is unknown. The main objective of this proposal is to determine the role of endothelial TSP-1 in protection against matrix proteolysis and barrier dysfunction following PA induced lung injury. Aim 1 will determine if TSP-1 is protective against the disruption of intercellular junctional complexes and barrier function in vitro. Aim 2 will determine if endothelial TSP-1 promotes vascular integrity in vivo. By improving our understanding of host protective mechanisms during pathogen triggered lung injury, this project may provide rationale for potential therapeutic targets in patients with ARDS. The proposed training plan will promote development of advanced laboratory skills including cell and tissue imaging, mouse genetics and in vivo models of infection-induced lung injury. Moreover, this project with provide the applicant with the opportunity to develop expertise as a physician-scientist under the close mentoring and involvement of dedicated sponsors in a robust research environment at the University of Pittsburgh.

项目摘要/摘要： 急性呼吸窘迫综合征（ARDS）是一种以呼吸系统快速进展为特征的综合征。 衰竭和难治性低氧血症，是全世界危重患者发病和死亡的主要原因。 ARDS 最常见的危险因素是严重细菌性肺炎，ARDS 患者也 经常受到包括下呼吸道感染（LRTI）在内的医院感染的困扰。假单胞菌属 铜绿假单胞菌 (PA) 是 ICU 常见的病原体，PA 肺炎与死亡率增加相关 与其他革兰氏阴性病原体相比，呼吸机相关性肺炎。 PA 已被证明会导致 广泛的蛋白水解损伤导致动物组织损伤和肺泡毛细血管屏障降解 模型，部分是通过分泌型外切蛋白酶（例如 LasB）实现的。金属蛋白酶 LasB 也已被证明 降解对屏障完整性很重要的连接蛋白。虽然病原体衍生的毒力因子已被 尽管具有广泛的特征，但有关宿主机制的知识有限，以防止 肺中的蛋白水解损伤和血管通透性。 Thrombospondin-1 (TSP-1) 是一种基质糖蛋白，具有 多种功能，包括调节炎症。 TSP-1 先前已被证明可以抑制宿主 衍生的丝氨酸蛋白酶和病原体衍生的外切蛋白酶 LasB。 TSP-1 由肺内皮细胞表达 细胞，但其在病原体引起的损伤中的作用尚不清楚。该提案的主要目标是确定 内皮 TSP-1 在 PA 诱导后防止基质蛋白水解和屏障功能障碍中的作用 肺损伤。目标 1 将确定 TSP-1 是否具有防止细胞间连接复合物破坏的保护作用 和体外屏障功能。目标 2 将确定内皮 TSP-1 是否促进体内血管完整性。经过 该项目提高了我们对病原体引发肺损伤期间宿主保护机制的理解 可能为 ARDS 患者的潜在治疗靶点提供理论基础。拟议的培训计划将 促进先进实验室技能的发展，包括细胞和组织成像、小鼠遗传学和体内 感染引起的肺损伤模型。此外，该项目为申请人提供了机会 在专门赞助商的密切指导和参与下发展作为医师科学家的专业知识 匹兹堡大学拥有强大的研究环境。