Elastase Injury of Pulmonary Extracellular Matrix

肺细胞外基质弹性蛋白酶损伤

• 批准号: 7368624
• 负责人: MATTHEW A NUGENT
• 金额: $ 40.63万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-12-01 至 2012-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7368624
• 关键词: Accounting; Animals; Apoptotic; Appearance; Atomic Force Microscopy; Back; Binding; Biochemical; Biological Assay; Cause of Death; Cells; Cessation of life; Chronic Obstructive Airway Disease; Circular Dichroism; Complex; Cultured Cells; Development; Disease; Docking; Elastases; Elastin; Endopeptidases; Enzyme Kinetics; Enzymes; Event; Exposure to; Extracellular Matrix; Fibroblasts; Fibronectins; GAG Gene; Gene Expression; Generations; Genetic; Growth Factor; Heparan Sulfate Proteoglycan; Histone Acetylation; Injury; Interleukin-13; Knockout Mice; Leukocyte Elastase; Lung; Measures; Mechanics; Mediating; Mediator of activation protein; Molecular; Nuclear; Numbers; Oxidative Stress; Pancreatic Elastase; Peptide Hydrolases; Process; Property; Protease Inhibitor; Protein Binding; Proteins; Proteoglycan; Pulmonary Emphysema; Role; Small Interfering RNA; Tropoelastin; United States; Vascular Endothelial Growth Factors; base; enzyme substrate; feeding; injury and repair; insight; lung injury; proteoglycan core protein; repaired; response; response to injury; syndecan; therapy development

DESCRIPTION (provided by applicant): In this project we will focus on defining the process of elastolytic injury of pulmonary cells and their extracellular matrix. In particular, we aim to identify the function of heparan sulfate proteoglycans (HSPGs) as regulators of elastolysis and modulators of the cellular response to injury. In addition, we will investigate the consequences of elastase injury to the extracellular matrix on vascular endothelial growth factor (VEGF) storage, release, and activity. Our previous studies have implicated proteoglycans as central mediators of elastase damage to pulmonary cells and matrix. Specifically, we have shown that HSPGs modulate growth factor storage, release, and transport within the matrix; moreover, we have also found that elastase-generated HSPG fragments feed back to inhibit elastase and that elastase leads to increased nuclear HSPGs, reduced histone acetylation, reduced tropoelastin expression, and release of VEGF fragments. We plan to expand our studies to focus on the function of released fragments of HSPGs, the specific HSPGs, syndecans 1 and 4, and VEGF. Our specific aims are to: 1) Identify the mechanisms and functional consequences of HSPG inhibition of elastase activity. 2) Define the mechanisms and functional consequences of elastase release of VEGF from pulmonary cells and matrix. 3) Determine the role of HSPGs in mediating the response of lung fibroblasts to elastase injury. We will use a combination of biochemical, molecular, and biophysical approaches in conjunction with cell culture and animal studies utilizing syndecan knock out mice to investigate the complex process of lung injury and repair. Ultimately, these studies will provide critical insight into the cascade of events initiated by elastolytic injury and the subsequent development of emphysema. Project Narrative: Chronic Obstructive Pulmonary Disease, which includes emphysema, is the fourth leading cause of death in the United States, accounting for more than 120,000 deaths in 2002. To develop effective therapies for this "quiet killer" a more complete understanding of the underlying cellular and molecular causes is required. In the present project, we propose to identify new critical components of how the lung responds, appropriately and inappropriately, to a specific type of injury (elastolysis) so that this information might provide insight toward the development of therapies that aim to assist lung repair and avoid disease.

描述（由申请人提供）：在此项目中，我们将重点定义肺细胞及其细胞外基质的弹性损伤过程。特别是，我们旨在确定硫酸乙酰肝素蛋白聚糖（HSPG）作为弹性的调节剂和细胞对损伤反应的调节剂的调节剂。此外，我们将研究弹性酶损伤对细胞外基质对血管内皮生长因子（VEGF）储存，释放和活性的后果。我们以前的研究已将蛋白聚糖作为对肺细胞和基质的弹性蛋白酶损伤的中心介体。具体而言，我们已经表明HSPG在矩阵内调节生长因子存储，释放和传输。此外，我们还发现，弹性酶产生的HSPG片段反馈至抑制弹性酶，弹性酶会导致核HSPG增加，组蛋白乙酰化降低，降低了tropoelastin的表达以及VEGF片段的释放。我们计划扩大研究，以关注HSPG，特定HSPG，Syndecans 1和4和VEGF的释放片段的功能。我们的具体目的是：1）确定HSPG抑制弹性酶活性的机制和功能后果。 2）定义从肺细胞和基质中VEGF释放的弹性酶释放的机制和功能后果。 3）确定HSPG在介导肺成纤维细胞对弹性酶损伤的反应中的作用。我们将结合生化，分子和生物物理方法与细胞培养和动物研究结合使用Syndecan敲除小鼠来研究肺损伤和修复的复杂过程。最终，这些研究将提供对弹性损伤引发的一系列事件和随后的肺气肿发展的批判性见解。项目叙述：包括肺气肿在内的慢性阻塞性肺部疾病是美国的第四个主要死亡原因，2002年占120,000多人的死亡。为这种“安静的杀手”开发有效的疗法，对这种“安静的杀手”有更全面的了解，需要对潜在的细胞和分子原因进行更全面的了解。在本项目中，我们建议确定肺部对特定类型的损伤类型（弹性）的反应的新关键组成部分，以便该信息可以为旨在帮助肺部修复和避免疾病的疗法的发展提供见解。