MECHANISMS OF LUNG BETA2AR FUNCTION IN TRANSGENIC MICE

转基因小鼠肺β2AR功能的机制

• 批准号: 6638086
• 负责人: Dennis W McGraw
• 金额: $ 13.13万
• 依托单位: UNIVERSITY OF CINCINNATI
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-05-04 至 2004-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6638086
• 关键词: beta adrenergic receptor; biological models; bronchomotion; cell type; genetically modified animals; inflammation; laboratory mouse; model design /development; receptor expression; respiratory epithelium; respiratory hypersensitivity; smooth muscle

Beta-agonists are potent bronchodilators that are frequently used in the treatment of bronchospastic lung diseases such as asthma. In the intact lung, Beta-agonists-mediated bronchodilation may result from the activation of Beta2ARs on a number of different cell types. However, the relative contribution of B2AR signaling from any one of these cell types is unclear. One potential approach to delineate these cell-specific effects in vivo is to use transgenesis to selectively over express, and thus activate, Beta2AR on specific cells. By using cell-specific promoters to target receptor expression, we propose to create multiple lines of transgenic mice that over express the Beta2AR in airway epithelium and in airway smooth muscle. Preliminary ex vivo and in vivo studies have indicated a significant Bets2AR- mediated communication between the epithelium and smooth muscle, in that over expression of the Beta2AR -mediated communication between the epithelium and smooth muscle, in airway epithelium decreases bronchial hyperactivity. Over expression on airway smooth muscle results in another phenotype, and this allows for assessment of each pathway with regard to Beta2AR regulation of airway function. Using a strategy that integrates molecular biological, pharmacological and physiological techniques, we propose to use such transgenic approaches to: (1) establish the role of epithelial cell Beta2ARs in regulating smooth muscle function; (2) selectively delineate Beta2AR signaling and function in airway smooth muscle; and (3) assess the effects of epithelial and smooth muscle Beta2AR over expression on airway inflammation and bronchial hyperresponsiveness. The candidate has completed a clinical fellowship in Pulmonary and Critical Care. During that time the candidate used basic molecular biological techniques to begin studies of Beta2AR gene expression. The candidate became interested in pursuing these studies further, and made the decision to move to the laboratory of Stephen Liggett at he University of Cincinnati Medical Center. After gaining more intensive laboratory experience, the candidate, mentor and secondary mentor (Jeffrey Whitsett) developed a plan utilizing transgenesis to study how Beta2AR on different cell types regulate airway function. The training plan proposed incorporates a variety of pharmacological and physiological techniques that represent a significant departure from the candidate's previous experience. Learning these additional techniques will serve to advance the candidate's skills to the level of autonomous investigator. To this aim, an environment has been created that is uniquely conducive to the candidate's training. This includes the mentor ship of Drs. Liggett and Whitsett, and an Advisory committee composed of established investigators with extensive experience in raining physicians scientists.

β-激动剂是有效的支气管扩张剂，经常用于治疗支气管疗法肺部疾病，例如哮喘。 在完整的肺中，β-激动剂介导的支气管扩张可能是由于β2AR在多种不同细胞类型上的激活而引起的。但是，这些细胞类型中任何一种的B2AR信号传导的相对贡献尚不清楚。描绘这些细胞特异性效应的一种潜在方法是使用转基因在特定细胞上选择性地过度表达并激活beta2ar。 通过使用细胞特异性启动子来靶向受体表达，我们建议创建多种转基因小鼠，这些小鼠在气道上皮和气道平滑肌中表达beta2AR。 初步的离体和体内研究表明，上皮和平滑肌之间存在明显的BETS2AR介导的交流，因为在气道上皮中，上皮和平滑肌之间的β2AR介导的交流过度表达可降低支气管的过度活跃性。 在气道平滑肌上的表达会导致另一种表型，这允许评估每种途径关于气道功能的beta2ar调节。 使用将分子生物学，药理和生理技术整合的策略，我们建议使用这种转基因方法来：（1）确定上皮细胞β2AR在调节平滑肌功能中的作用； （2）在气道平滑肌中有选择地描述beta2ar信号传导和功能； （3）评估上皮和平滑肌beta2ar对表达对气道炎症和支气管高反应性的影响。候选人已完成肺和重症监护的临床研究金。 在此期间，候选者使用基本的分子生物学技术开始研究β2AR基因表达。候选人对进一步进行这些研究感兴趣，并决定搬到辛辛那提大学医学中心的斯蒂芬·利格特（Stephen Liggett）实验室。 在获得更密集的实验室经验之后，候选人，导师和次要导师（Jeffrey Whitsett）制定了一项计划，利用转基因研究beta2AR如何调节气道功能。培训计划提出的培训计划结合了各种药理和生理技术，这些技术与候选人以前的经历有很大的不同。 学习这些其他技术将有助于将候选人的技能提高到自主研究者的水平。 为此，创造了一个环境，该环境极为有利于候选人的培训。 这包括Drs的导师船。 Liggett和Whitsett，以及由既定的研究人员组成的咨询委员会，在雨医师科学家方面拥有丰富经验。