Alveolar epithelial B2-adrenergic receptors

肺泡上皮 B2 肾上腺素能受体

基本信息

批准号：
7431602
负责人：
Phillip H Factor
金额：
$ 34.12万
依托单位：
COLUMBIA UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2001
资助国家：
美国
起止时间：
2001-07-01 至 2010-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7431602
关键词：
A kinase anchoring protein Accounting Active Biological Transport Acute Acute Lung Injury Adaptor Signaling Protein Address Adenylate Cyclase Adrenergic Agonists Adrenergic Receptor Affect Agonist Alveolar Alveolus Animal Model Binding Binding Proteins Cardiac Myocytes Carrier Proteins Cell membrane Cells Complement Complex Condition Cyclic AMP Cyclic AMP-Dependent Protein Kinases Cystic Fibrosis Transmembrane Conductance Regulator Cytoskeleton Data Databases Disruption Edema Epithelial Epithelial Cells Exposure to GTP-Binding Proteins Generations Goals Hour Human Immunoprecipitation In Vitro Injury Ion Transport Ligands Liquid substance Lung Macromolecular Complexes Mass Spectrum Analysis Measurement Measures Mediating Membrane Metabolic Clearance Rate Modeling Molecular Mus Na(+)-K(+)-Exchanging ATPase Organ Oxygen Patients Peptides Phosphorylation Physiological Preparation Procaterol Production Protein Overexpression Proteins Pulmonary Edema Purpose Receptor Activation Regulation Reporting Research Personnel Resolution Signal Transduction Speed Structure Testing Therapeutic alveolar epithelium apical membrane epithelial Na+ channel improved injured lung injury phosphoric diester hydrolase programs protein function protein protein interaction receptor receptor upregulation research study response scaffold

项目摘要

DESCRIPTION (provided by applicant): The clearance of pulmonary edema fluid occurs by means of active Na+ transport by alveolar epithelial cells. It is widely accepted that the coordinated function of Na+ and CI- channels in the apical membrane and Na,K-ATPase in the basolateral aspect of alveolar epithelial cells creates a transepithelial osmotic gradient that causes fluid to exit the alveolus. It has been observed in animal models and humans that in some types of lung injury active Na+ transport is impaired. Substantial experimental data suggests that beta2-adrenergic agonists accelerate active Na+ transport and speed edema resolution. These data offer the possibility that beta2-adrenergic agonists may be useful for the treatment of pulmonary edema. The overall goal of our experimental program is to improve our understanding of how beta2-adrenergic receptors (beta2AR) regulate alveolar active Na+ transport. We have reported that mice with no beta1 or beta2 adrenergic receptors have normal total lung cAMP levels but are unable to upregulate active Na+ transport in response to excess alveolar fluid. This finding led us to consider that beta2AR regulation of active Na+ transport requires more than cAMP production. It has recently been noted that the a2AR interacts with scaffold and adaptor proteins that are in close proximity to beta2AR effector molecules such as PKA and CFTR and, anchor it to the sub-membrane cytoskeleton. These protein-protein interactions allow for compartmentalized signaling and tight regulation of beta2AR function. In preparation for this competitive renewal we conducted preliminary immunoprecipitation studies that suggest that alveolar epithelial (2ARS form macromolecular complexes with other transport proteins. This new data led us to hypothesize that: beta2AR regulation of alveolar epithelial active transport occurs via highly regulated interactions with scaffold and/or adaptor proteins. If confirmed, this hypothesis would support a new paradigm where beta2AR regulation of alveolar active Na+ transport is dependent not only on cAMP generation but also formation of a macromolecular regulatory complex at the cell membrane. To address this hypothesis we have formulated the following three specific aims: Aim 1: Determine if beta2AR-scaffold/adaptor protein interactions are necessary for regulation of beta2AR sensitive active Na+ transport in alveolar epithelial cells in vitro. Aim 2: Ascertain if beta2AR -scaffold interactions are necessary for beta2AR regulation of active Na+ transport in normal lungs. Aim 3: Determine if acute lung injury alters beta2AR -scaffold/adaptor protein interactions in mouse lung. The focused studies in this competitive renewal application are structured to define which scaffold and adaptor proteins alveolar beta2AR interacts with, which are required for regulation of active Na+ transport, and if these proteins are affected by acute lung injury. These studies offer an opportunity to expand our understanding of the mechanisms by which the alveolar epithelium regulates active Na+ transport. It is hoped that our studies will identify injury-induced alterations in beta2AR protein-protein interactions that would be amenable to therapeutic manipulation for purposes of speeding resolution of pulmonary edema in the millions of patients with acute pulmonary edema each year.

描述（由申请人提供）：肺水肿液的清除是通过肺泡上皮细胞主动转运 Na+ 来实现的。人们普遍认为，顶膜中的 Na+ 和 CI- 通道以及肺泡上皮细胞基底外侧的 Na,K-ATP 酶的协调功能会产生跨上皮渗透梯度，导致液体流出肺泡。在动物模型和人类中观察到，在某些类型的肺损伤中，活性 Na+ 转运受到损害。大量实验数据表明，β2-肾上腺素能激动剂可加速活性 Na+ 转运并加速水肿消退。这些数据表明β2-肾上腺素能激动剂可能有助于治疗肺水肿。我们实验计划的总体目标是提高我们对β2-肾上腺素能受体（β2AR）如何调节肺泡活性Na+转运的理解。我们报道过，没有β1或β2肾上腺素能受体的小鼠具有正常的肺总cAMP水平，但无法响应过量的肺泡液而上调活性Na+转运。这一发现使我们认为 beta2AR 对活性 Na+ 转运的调节需要的不仅仅是 cAMP 的产生。最近注意到，a2AR 与支架和接头蛋白相互作用，这些蛋白与 β2AR 效应分子（例如 PKA 和 CFTR）非常接近，并将其锚定到亚膜细胞骨架上。这些蛋白质-蛋白质相互作用允许划分信号传导并严格调节 beta2AR 功能。为了准备这种竞争性更新，我们进行了初步的免疫沉淀研究，结果表明肺泡上皮 (2ARS) 与其他转运蛋白形成大分子复合物。这一新数据使我们推测：β2AR 对肺泡上皮主动转运的调节是通过与支架和支架的高度调节的相互作用发生的。 /或接头蛋白。如果得到证实，这一假设将支持一种新的范式，其中β2AR对肺泡活性Na+转运的调节不仅依赖于cAMP的生成，还依赖于细胞膜上大分子调节复合物的形成。为了解决这一假设，我们制定了以下三个具体目标：目标 1：确定 β2AR-支架/适配器蛋白相互作用是否对于调节体外肺泡上皮细胞中 β2AR 敏感的活性 Na+ 转运是必要的。目标 2：确定 β2AR -支架相互作用对于正常肺中活性 Na+ 转运的 β2AR 调节是否是必需的。目标 3：确定急性肺损伤是否改变小鼠肺中 β2AR 支架/接头蛋白的相互作用。这项竞争性更新应用的重点研究旨在确定肺泡 β2AR 与哪些支架和接头蛋白相互作用，这些蛋白是调节活性 Na+ 转运所必需的，以及这些蛋白是否受到急性肺损伤的影响。这些研究为扩大我们对肺泡上皮调节活性 Na+ 转运的机制的理解提供了机会。希望我们的研究能够确定损伤引起的β2AR蛋白-蛋白相互作用的改变，这些改变适合于治疗操作，以加速每年数百万急性肺水肿患者的肺水肿消退。

项目成果

期刊论文数量（6）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Interdependency of beta-adrenergic receptors and CFTR in regulation of alveolar active Na+ transport.

β-肾上腺素能受体和 CFTR 在调节肺泡活性钠转运中的相互依赖性。

DOI：
发表时间：
2005-05-13
期刊：
影响因子：
0
作者：
Mutlu, Gokhan M;Adir, Yochai;Jameel, Mohammed;Akhmedov, Alexander T;Welch, Lynn;Dumasius, Vidas;Meng, Fan Jing;Zabner, Joseph;Koenig, Craig;Lewis, Erin Rachel;Balagani, Rajesh;Traver, Geri;Sznajder, Jacob I;Factor, Phillip
通讯作者：
Factor, Phillip

In vivo molecular imaging characterizes pulmonary gene expression during experimental lung transplantation.

体内分子成像表征实验性肺移植过程中肺基因表达的特征。

DOI：
发表时间：
2005-06
期刊：
影响因子：
0
作者：
Dharmarajan, Sekhar;Hayama, Makio;Kozlowski, James;Ishiyama, Takaaki;Okazaki, Mikio;Factor, Phillip;Patterson, G Alexander;Schuster, Daniel P
通讯作者：
Schuster, Daniel P

In vivo timing of onset of transgene expression following adenoviral-mediated gene transfer.

腺病毒介导的基因转移后转基因表达开始的体内时间。

DOI：
发表时间：
2003-04-10
期刊：
影响因子：
3.7
作者：
Dumasius, Vidas;Jameel, Mohammed;Burhop, James;Meng, Fan Jing;Welch, Lynn C;Mutlu G, Gökhan M;Factor, Phillip
通讯作者：
Factor, Phillip

Upregulation of alveolar epithelial active Na+ transport is dependent on beta2-adrenergic receptor signaling.

肺泡上皮活性 Na 转运的上调依赖于 β2 肾上腺素能受体信号传导。

DOI：
发表时间：
2004-04-30
期刊：
影响因子：
0
作者：
Mutlu, Gokhan M;Dumasius, Vidas;Burhop, James;McShane, Pamela J;Meng, Fan Jing;Welch, Lynn;Dumasius, Andrew;Mohebahmadi, Nima;Thakuria, Gloria;Hardiman, Karen;Matalon, Sadis;Hollenberg, Steven;Factor, Phillip
通讯作者：
Factor, Phillip

Alveolar epithelial beta 2-adrenergic receptors: their role in regulation of alveolar active sodium transport.

肺泡上皮β2-肾上腺素能受体：它们在调节肺泡活性钠转运中的作用。