AREA Program: In Vitro Modeling of Smoke Induced Loss of Lung Barrier Function

AREA 计划：烟雾引起的肺屏障功能丧失的体外建模

• 批准号: 7458055
• 负责人: CINDY M KNALL
• 金额: $ 19.77万
• 依托单位: UNIVERSITY OF ALASKA ANCHORAGE
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-05-01 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7458055
• 关键词: Address; Air; Biological Models; Build-it; Cell physiology; Cells; Chronic; Chronic Obstructive Airway Disease; Cigarette; Condition; Country; Development; Environment; Epithelial; Epithelial Cells; Epithelium; Generations; Goals; In Vitro; Institution; Liquid substance; Lung; Lung diseases; Marketing; Methodology; Myosin ATPase; Myosin Light Chain Kinase; Outcome; Permeability; Range; Regulation; Research; Rho-associated kinase; Signal Transduction Pathway; Smoke; Smoker; Structure of respiratory epithelium; Students; System; Techniques; Testing; Therapeutic; Therapeutic Intervention; Tight Junctions; United States; Work; airway epithelium; base; cigarette smoke-induced; cigarette smoking; cigarette smoking; cost; expectation; improved; in vitro Model; in vivo; in vivo Model; innovation; insight; killings; programs

DESCRIPTION (provided by applicant): Cigarette smoking causes 80-90% of all cases of chronic obstructive pulmonary disease, COPD, which annually kills nearly 125,000 people in the United States and costs this country $37.2 billion. Cigarette smoke rapidly increases the permeability of airway epithelium, especially in those with COPD. The barrier function of an intact epithelium is maintained by tight junctions between adjacent cells. Little is known about the mechanisms underlying tight junction function in respiratory epithelium. Our long-range goal is to define the mechanisms responsible for cigarette smoke induced alterations in lung epithelial cell function in order to develop effective therapeutic strategies to inhibit this altered function. The objective of this proposal is to refine the methodology of in vitro air-liquid interface exposure of lung epithelial cells to whole, fresh cigarette smoke to improve the identification of regulatory mechanisms controlling smoke altered tight junction function. The central hypothesis of this proposal is that smoke from current market relevant cigarettes alters tight junction function because of altered cytoskeletal regulation by Rho kinase (ROCK) and myosin light chain kinase (MLCK). This hypothesis is based on strong preliminary findings obtained using a first generation in vitro model system. The rationale for this proposal is that the development of an improved in vitro exposure system that more closely reflects the current in vivo condition in smokers will allow mechanisms that control lung epithelial cell function to be defined, resulting in the identification of potential targets for therapeutic interventions. The central hypothesis will be tested and the objective accomplished through two specific aims: 1) refine an in vitro air-liquid interface exposure system that models in vivo lung exposure; 2) determine the extent to which ROCK and MLCK contribute to the reversible loss of tight junction function in lung epithelial cells exposed to cigarette smoke. The proposed work is innovative because it builds on a recently developed technique to expose lung epithelial cells in vitro under conditions that mimic the exposure conditions within the in vivo lung. Our expectation is that this approach will establish that ROCK and MLCK through their regulation of myosin contribute to the regulation of tight junction function in lung epithelial cells. The outcome is significant because it will begin to define the signal transduction pathways that regulate cigarette smoke induced loss of tight junction function in lung epithelium, and will provide insights for the development of strategies to control the loss of epithelial integrity. The proposed study fulfills the AREA program objectives of meritorious research to engage students in the research endeavor and to improve the research environment at this primarily undergraduate institution. PROJECT NARRATIVE. Cigarette smoking causes chronic obstructive pulmonary disease (COPD) which annually kills nearly 125,000 people in the US and costs this country $37.2 billion. Cigarette smoke causes the lungs in people to become leaky, especially in those with COPD. This project will begin to address how cigarette smoke makes lung cells leaky.

描述（由申请人提供）：吸烟会导致所有慢性阻塞性肺部疾病的80-90％COPD，每年杀死美国近125,000人，损失该国372亿美元。香烟烟雾迅速增加了气道上皮的渗透性，尤其是在患有COPD的人中。完整上皮的屏障功能由相邻细胞之间的紧密连接保持。关于呼吸上皮的紧密连接功能的机制知之甚少。我们的远程目标是定义负责香烟烟雾引起的肺上皮细胞功能改变的机制，以制定有效的治疗策略以抑制这种改变的功能。该提案的目的是完善肺上皮细胞在整个新鲜香烟烟雾中暴露于体外空气界面的方法，以改善控制烟雾改变紧密连接功能的调节机制。该提案的中心假设是，由于Rho激酶（ROCK）和肌球蛋白轻链激酶（MLCK）的细胞骨架调节改变了，当前市场相关的香烟的烟雾改变了紧密的连接功能。该假设基于使用第一代体外模型系统获得的强烈初步发现。该提案的理由是，改进的体外暴露系统的开发更加紧密地反映了吸烟者中当前的体内状态，将允许定义控制肺上皮细胞功能的机制，从而确定治疗干预措施的潜在靶标。将测试中央假设，并通过两个特定目的来实现目标：1）改进体外肺暴露模型的体外空气界面暴露系统； 2）确定岩石和MLCK在暴露于香烟烟雾中的肺上皮细胞中造成紧密连接功能的可逆损失的程度。提出的工作具有创新性，因为它建立在最近开发的技术基础上，该技术在模仿体内肺内暴露条件的条件下，在体外暴露肺上皮细胞。我们的期望是，这种方法将通过调节肌球蛋白的调节来确定岩石和MLCK有助于调节肺上皮细胞中紧密连接功能。结果很重要，因为它将开始定义调节香烟烟雾引起的肺上皮连接功能丧失的信号转导途径，并将为建立控制上皮完整性丧失的策略提供见解。拟议的研究符合有功研究的领域计划目标，使学生参与研究工作，并改善该主要本科机构的研究环境。项目叙述。吸烟会引起慢性阻塞性肺部疾病（COPD），每年杀死美国近125,000人，而该国损失了372亿美元。香烟烟使人们的肺部漏水，尤其是患有COPD的肺部。该项目将开始解决香烟烟雾如何使肺部细胞漏水。