The Role of Caveolins in Lung Inflammation

小窝蛋白在肺部炎症中的作用

• 批准号: 8235009
• 负责人: Christina Maria Pabelick
• 金额: $ 33.66万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8235009
• 关键词: Address; Affect; Agonist; Airway Resistance; Area; Asthma; Biochemical; Caveolae; Caveolins; Cell membrane; Cells; Cellular Structures; Chronic Bronchitis; Cyclic ADP-Ribose; Data; Development; Disease; Fluorescence; Foundations; Generations; Goals; Health; Human; Imaging Techniques; In Vitro; Inflammation; Inflammatory; Knock-out; Knockout Mice; Lead; Link; Lipids; Lung; Lung Inflammation; MAP Kinase Gene; Maintenance; Measures; Mediating; Membrane Proteins; Mitogen-Activated Protein Kinases; Modeling; Molecular Biology; Mus; Muscarinic M3 Receptor; Ovalbumin; Pathway interactions; Pharmacology; Phosphorylation; Physiological; Proteins; Regulation; Reporting; Research; Rho-associated kinase; Role; STIM1 gene; Sarcoplasmic Reticulum; Second Messenger Systems; Shapes; Shortness of Breath; Signal Transduction; Small Interfering RNA; Smooth Muscle Myocytes; Structural Protein; Structure; TNF gene; TNFRSF1A gene; Techniques; Tissues; Transfection; Tumor Necrosis Factor Receptor; Tumor Necrosis Factor-Beta; Tumor Necrosis Factor-alpha; Viral; Work; airway hyperresponsiveness; airway inflammation; base; caveolin 1; caveolin-2; cell type; cytokine; flasks; genetic regulatory protein; in vivo; inhibitor/antagonist; laser capture microdissection; methacholine; migration; mouse model; new therapeutic target; protein expression; receptor; respiratory smooth muscle; response; rho; scaffold; second messenger

DESCRIPTION (provided by applicant): Exaggerated airway narrowing and inflammation are hallmarks of clinically-important diseases such as asthma. Airway tone, largely determined by intracellular Ca2+ ([Ca2+]i) (mediated by Ca2+ influx and sarcoplasmic (SR) Ca2+ release) and Ca2+ sensitivity for force generation of airway smooth muscle (ASM), is enhanced by pro-inflammatory cytokines such as TNFa. In this regard, the role of caveolae and caveolins in the lung is an exciting and emerging area of research. Caveolae are small, uncoated flask-shaped invaginations of the plasma membrane (PM) of most cell types, and contain structural proteins called caveolins (caveolin-1, -2 and/or -3). Recent studies including our own have demonstrated that normal human ASM expresses caveolae and caveolin-1, with specific proteins involved in [Ca2+]i and force regulation co-localizing with caveolin-1. Preliminary studies show that airway inflammation (either in vitro following TNFa exposure or in vivo in a mouse model) increases caveolin-1 expression and enhances caveolar regulation of ASM contractility, suggesting that overall, caveolin-1 is associated with or promotes ASM contractility. The long term goal of the proposed studies is to understand the mechanisms by which caveolae and caveolins regulate ASM contractility under normal conditions and with airway disease. Ca2+ The overall hypothesis is that caveolae facilitate PM- SR interactions, thus influencing [Ca2+]i and force in ASM. In addition, we hypothesize that airway inflammation increases caveolae and caveolin-1 expression, and thus enhances PM-SR interactions, leading to increased [Ca2+]i and force in ASM. The novelty of our studies lies in linking previous anatomical and biochemical evidence for caveolin-1 in ASM to a physiological role in regulation of ASM contractility. Overall Approach: In vitro studies with human ASM cells and tissues in Aim 1 will examine mechanisms that can be modulated by caveolin-1 with agonist (ACh) stimulation under normal conditions, focusing on agonist receptors, second messengers (PLC/IP3, cADPR), regulation of store-operated Ca2+ entry (SOCE) by (STIM1/Orai1), and RhoA (Ca2+ sensitization). Mechanisms underlying altered caveolin-1 expression (Aim 2) and caveolin-1 contribution to enhanced ASM contractility (Aim 3) with inflammation (induced by TNFa) will then be examined, focusing on specific mechanisms (TNFR1, PTRF, MAPK, NF-?B). Finally in Aim 4, in vitro data will be integrated at the in vivo level using an ovalbumin (OVA) model of airway hyperresponsiveness in wildtype (WT) and caveolin-1 knockout (KO) mice. The Specific Aims of this proposal are: 1) To determine mechanisms by which caveolae/caveolin-1 regulate [Ca2+]i and force responses of human ASM to agonist 2 ) To determine mechanisms by which cytokine (TNFa) stimulation alters caveolin-1 expression in human ASM 3) To determine the mechanisms by which caveolin-1 contributes to TNFa-induced enhancement of [Ca2+]i and force in human ASM; 4) To determine the role of caveolin-1 in airway hyperresponsiveness in vivo. PUBLIC HEALTH RELEVANCE: There is increasing recognition that abnormalities in airway smooth muscle contractility (exacerbated by inflammation) contribute to exaggerated airway narrowing and accompanying shortness of breath in clinically- important diseases such as asthma and chronic bronchitis. In this regard, the potential role of cellular structures called caveolae and their constituent caveolin proteins in regulation of airway contractility is an exciting and emerging area of research. By establishing the role of caveolae and caveolins in airway narrowing with or without inflammation, the proposed studies will the foundation for better understanding of airway diseases, and potential development of new therapeutic targets.

描述（由申请人提供）：夸张的气道狭窄和炎症是临床上最重要的疾病（例如哮喘）的标志。气道音调在很大程度上由细胞内Ca2+（[Ca2+] I）确定（由Ca2+流入和肌胞浆（SR）Ca2+释放介导）和CA2+敏感性，用于产生气道平滑肌（ASM）的力，通过诸如TNFA等促炎细胞因子（例如TNFA）增强了气道平滑肌（ASM）。在这方面，小窝和小窝蛋白在肺中的作用是一个令人兴奋的研究领域。小窝是大多数细胞类型的质膜（PM）的小烧瓶形状的小烧瓶形，并且包含称为小窝蛋白（Caveolin -1，-2和/或-3）的结构蛋白。包括我们自己的包括我们自己的最新研究表明，正常的人ASM表达小窝和小窝蛋白-1，特定的蛋白质参与[Ca2+] i，而强力调节与Caveolin-1共定位。初步研究表明，气道炎症（在TNFA暴露后的体外或小鼠模型中的体内体外）会增加小窝蛋白-1的表达，并增强对ASM收缩力的洞穴调节，这表明Caveolin-1与ASM的总体相关或促进了ASM的收缩力。拟议研究的长期目标是了解小窝和小窝蛋白在正常条件和气道疾病中调节ASM收缩力的机制。 CA2+总体假设是小窝促进PM-SR相互作用，从而影响[Ca2+] I和ASM中的力。此外，我们假设气道炎症会增加小窝和小窝蛋白1的表达，从而增强PM-SR相互作用，从而导致[Ca2+] I和ASM的力增加。我们研究的新颖性在于将Caveolin-1在ASM中的先前解剖学和生化证据联系起来，与ASM收缩性调节中的生理作用联系起来。 Overall Approach: In vitro studies with human ASM cells and tissues in Aim 1 will examine mechanisms that can be modulated by caveolin-1 with agonist (ACh) stimulation under normal conditions, focusing on agonist receptors, second messengers (PLC/IP3, cADPR), regulation of store-operated Ca2+ entry (SOCE) by (STIM1/Orai1), and RhoA (Ca2+ sensitization).然后，将检查改变小窝蛋白-1表达（AIM 2）和小窝蛋白1对增强ASM收缩力（AIM 3）的机制（AIM 3），然后检查具有特定机制（TNFR1，PTRF，MAPK，NF--？B）。最后，在AIM 4中，使用野生型（WT）和Caveolin-1敲除（KO）小鼠的气道高反应性的椭圆形模型在体内水平上进行体外数据。 The Specific Aims of this proposal are: 1) To determine mechanisms by which caveolae/caveolin-1 regulate [Ca2+]i and force responses of human ASM to agonist 2 ) To determine mechanisms by which cytokine (TNFa) stimulation alters caveolin-1 expression in human ASM 3) To determine the mechanisms by which caveolin-1 contributes to TNFa-induced enhancement of [Ca2+]i and人类ASM的武力； 4）确定Caveolin-1在体内气道高反应性中的作用。公共卫生相关性：越来越多的认识是，气道平滑肌收缩性异常（因炎症而加剧）有助于夸张的气道变窄，并伴随着临床上重要疾病（例如哮喘和慢性支气管炎）的呼吸急促。在这方面，称为Caveolae的细胞结构的潜在作用及其成分的小窝蛋白在调节气道收缩力中的潜在作用是一个令人兴奋的研究领域。通过建立小窝和小窝蛋白在气道中的作用，无论有或没有炎症，拟议的研究将为更好地理解气道疾病以及潜在的新治疗靶标的发展。