Airway and Lung Vascular Remodeling in COPD

慢性阻塞性肺病 (COPD) 中的气道和肺血管重塑

• 批准号: 8967091
• 负责人: YUNCHAO SU
• 金额: --
• 依托单位: CHARLIE NORWOOD VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-10-01 至 2018-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8967091
• 关键词: Animal Model; Binding; Calcium; Calpain; Caring; Caspase; Cause of Death; Cell Proliferation; Cells; Cellular biology; Cessation of life; Chronic; Chronic Obstructive Airway Disease; Cleaved cell; Collagen; Data; Development; Epithelial; Family; Genes; Golgi Apparatus; Health Care Costs; Hydrogen Peroxide; Hypoxia; Inflammatory; Knock-out; Knockout Mice; Lead; Life; Lung; Lung diseases; Mammalian Cell; Mediating; Mediator of activation protein; Methods; Modeling; Monocrotaline; Morbidity - disease rate; Outcome; Pathogenesis; Pathway interactions; Play; Population; Prevalence; Prevention; Process; Production; Proteins; Proteolysis; Pulmonary Heart Disease; Pulmonary Hypertension; Pulmonary artery structure; Rattus; Reactive Oxygen Species; Reporting; Risk Factors; Role; Signal Pathway; Signal Transduction; Small Interfering RNA; Smooth Muscle; Smooth Muscle Myocytes; Source; TGF-beta type I receptor; Testing; Time; Tobacco smoke; Tobacco use; Transforming Growth Factors; United States; Vascular Smooth Muscle; Vascular remodeling; Veterans; Work; arteriole; bronchial artery; calpain inhibitor; calpastatin; improved; inhibitor/antagonist; innovation; insight; interdisciplinary approach; novel; overexpression; prevent; public health relevance; receptor; research study

DESCRIPTION (provided by applicant): Tobacco use and prevalence of chronic obstructive pulmonary disease (COPD) are significantly higher among veterans than non-veteran population. Airway and pulmonary vascular remodeling is the most critical process in the pathogenesis of COPD. Large amounts of reactive oxygen species (ROS) are released from activated inflammatory cells and other lung cells in COPD or from tobacco smoke (TS) per se. ROS are implicated in the development of airway and pulmonary vascular remodeling. However, little has been known about how ROS and TS lead to these pathological alterations. We have found that ROS and TS activates calpain in bronchial and pulmonary artery smooth muscle cells (BSMCs and PASMCs) and that calpain mediates cell proliferation and collagen-I synthesis in PASMCs. Calpain is a family of calcium-dependent non- lysosomal neutral cysteine endopeptidases that act via limited proteolysis of substrate proteins in mammalian cells, including BSMCs and PASMCs. We have recently reported for the first time that global inhibition of calpain prevents pulmonary vascular remodeling induced by chronic hypoxia and monocrotaline. In preliminary experiments, we found that the specific calpain inhibitor, MDL28170, prevented ROS-induced increases in cell proliferation and protein levels of collagen-I in BSMCs and PASMCs. Transforming growth factor (TGF)/Smad pathway have been shown to play important roles in airway and pulmonary vascular remodeling. We have reported that calpain activates TGF1 by cleaving latent TGF1. This proposal is to study a novel hypothesis that calpain mediates ROS- and TS-induced cell proliferation and collagen synthesis in airway and pulmonary vascular smooth muscle cells via activation of intracellular TGF1 in TS-induced COPD. The protein and activity of calpain, cell proliferation and collagen synthesis will be determined in BSMCs and PASMCs with or without treatment of H2O2 and TS. Calpain activity will be manipulated using specific inhibitor, siRNA and over-expression of calpastatin genes. A novel intracrine TGF1 pathway will be studied in H2O2- and TS-treated cells. Moreover, the role of calpain in airway and pulmonary vascular remodeling will be evaluated in a COPD animal model using innovative inducible global and smooth muscle-specific calpain knockout mouse line. Finally, calpain will be pharmacologically targeted using a specific calpain inhibitor MDL28170 in a TS-induced COPD rat model. This proposal is novel because it will identify calpain as a mediator in airway and pulmonary vascular remodeling and calpain serves this role by activating intracellular TGF1 in BSMCs and PASMCs. A better understanding of the mechanistic insight will provide a strong rationale for manipulating calpain activities in the treatment of lung diseases such as COPD, pulmonary hypertension and cor pulmonale. Completion of this project will help find novel treatments for COPD and in turn improve the life and care of veterans.

描述（由申请人提供）： 退伍军人的慢性阻塞性肺疾病（COPD）的使用和患病率明显高于非退伍军人。气道和肺血管重塑是COPD发病机理中最关键的过程。大量活性氧（ROS）是从活化的炎症细胞和COPD中的其他肺细胞或烟草烟雾（TS）中释放的。 ROS与气道和肺血管重塑的发展有关。但是，关于ROS和TS如何导致这些病理改变的知识知之甚少。我们发现ROS和TS激活了支气管和肺动脉平滑肌细胞（BSMC和PASMC）中的钙蛋白酶，并且Calpain介导PASMC中的细胞增殖和胶原蛋白I合成。钙蛋白酶是钙依赖性的非溶酶体中性半胱氨酸内肽酶的家族，可通过有限的乳腺细胞中底物蛋白（包括BSMC和PASMC）起作用。我们最近首次报道了全球抑制钙蛋白酶可防止慢性缺氧和单蛋白引起的肺血管重塑。在初步实验中，我们发现特异性的钙蛋白酶抑制剂MDL28170阻止了ROS诱导的BSMC和PASMC中胶原蛋白I的细胞增殖和蛋白质水平的增加。转化生长因子（TGF）/SMAD途径已显示在气道和肺血管重塑中起重要作用。我们报告说，钙蛋白酶通过切割潜在的TGF1来激活TGF1。该建议是研究一种新的假设，该假设可通过在TS诱导的COPD中激活气道和肺血管平滑肌细胞中ROS-和TS诱导的细胞增殖和胶原蛋白合成。钙蛋白酶，细胞增殖和胶原蛋白合成的蛋白质和活性将在带有H2O2和TS处理的BSMC和PASMC中确定。钙蛋白酶活性将使用特异性抑制剂，siRNA和钙蛋白酶基因过表达进行操纵。将在H2O2和TS处理的细胞中研究一种新型的内部TGF1途径。此外，将使用创新的诱导性全球和平滑肌特异性的Calpain敲除小鼠系列在COPD动物模型中评估CALPAIN在气道和肺血管重塑中的作用。最后，在TS诱导的COPD大鼠模型中，使用特定的Calpain抑制剂MDL28170将钙蛋白酶用于药理学靶向。该建议之所以新颖，是因为它可以将钙蛋白酶鉴定为气道和肺血管重塑中的介体，而钙蛋白酶通过在BSMC和PASMC中激活细胞内TGF1来发挥这种作用。更好地了解机械洞察力将为操纵钙蛋白酶活性在治疗肺部疾病（例如COPD，肺动脉高压和COR肺癌）方面提供强大的理由。该项目的完成将有助于找到针对COPD的新型治疗方法，进而改善退伍军人的生活和照顾。