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）： 退伍军人中的烟草使用率和慢性阻塞性肺病（COPD）患病率显着高于非退伍军人。气道和肺血管重塑是COPD发病机制中最关键的过程。 COPD 中激活的炎症细胞和其他肺细胞或烟草烟雾 (TS) 本身会释放大量活性氧 (ROS)。 ROS 参与气道和肺血管重塑的发展。然而，人们对 ROS 和 TS 如何导致这些病理改变知之甚少。我们发现 ROS 和 TS 激活支气管和肺动脉平滑肌细胞（BSMC 和 PASMC）中的钙蛋白酶，并且钙蛋白酶介导 PASMC 中的细胞增殖和 I 型胶原蛋白合成。钙蛋白酶是钙依赖性非溶酶​​体中性半胱氨酸内肽酶家族，其通过哺乳动物细胞（包括 BSMC 和 PASMC）中底物蛋白的有限蛋白水解发挥作用。我们最近首次报道了钙蛋白酶的整体抑制可以防止慢性缺氧和野百合碱引起的肺血管重塑。在初步实验中，我们发现特定的钙蛋白酶抑制剂 MDL28170 可阻止 ROS 诱导的 BSMC 和 PASMC 中细胞增殖和 I 型胶原蛋白水平的增加。转化生长因子（TGF）/Smad 通路已被证明在气道和肺血管重塑中发挥重要作用。我们报道过钙蛋白酶通过裂解潜在的 TGF1 来激活 TGF1。本提案旨在研究一个新的假设，即在 TS 诱导的 COPD 中，钙蛋白酶通过激活细胞内 TGF1 介导 ROS 和 TS 诱导的气道和肺血管平滑肌细胞的细胞增殖和胶原蛋白合成。在经过或未经 H2O2 和 TS 处理的 BSMC 和 PASMC 中，将测定钙蛋白酶的蛋白质和活性、细胞增殖和胶原合成。使用特定抑制剂、siRNA 和钙蛋白酶抑制素基因的过表达来控制钙蛋白酶活性。将在 H2O2 和 TS 处理的细胞中研究一种新的内分泌 TGF1 途径。此外，将使用创新的诱导性整体和平滑肌特异性钙蛋白酶敲除小鼠系在慢性阻塞性肺病动物模型中评估钙蛋白酶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，将使用特定的钙蛋白酶抑制剂 MDL28170 在药理上靶向钙蛋白酶。该提议是新颖的，因为它将确定钙蛋白酶作为气道和肺血管重塑的介质，并且钙蛋白酶通过激活 BSMC 和 PASMC 中的细胞内 TGF1 来发挥这一作用。更好地理解这一机制将为控制钙蛋白酶活性治疗慢性阻塞性肺病、肺动脉高压和肺心病等肺部疾病提供强有力的理论基础。该项目的完成将有助于找到治疗慢性阻塞性肺病的新疗法，从而改善退伍军人的生活和护理。