Molecular signal transduction of cAMP compartments

cAMP 区室的分子信号转导

基本信息

批准号：
9189627
负责人：
RENNOLDS S OSTROM
金额：
$ 26.56万
依托单位：
CHAPMAN UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-01-01 至 2018-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9189627
关键词：
A kinase anchoring protein Adenylate Cyclase Alpha Cell Asthma Biochemical Biological Assay Biological Models Cell Differentiation process Cell Shape Cell membrane Cell model Cell physiology Cells Chronic Obstructive Airway Disease Congestive Heart Failure Coronary Arteriosclerosis Coupled Coupling Cyclic AMP Cyclic AMP Receptors Data Disease Enzymes Fibroblasts Fluorescence Resonance Energy Transfer Future G-Protein-Coupled Receptors Goals Hormone Receptor Human Hypertension Individual Interleukin-6 Kinetics Lead Location Membrane Microdomains Molecular Monitor Muscle Cells Nature Neurotransmitter Receptor Physiological Positioning Attribute Production Protein Isoforms Proteins Pulmonary Fibrosis Receptor Signaling Role Second Messenger Systems Sensitivity and Specificity Signal Transduction Somatostatin Source Stroke Study models Therapeutic Time base cell type design differential expression interdisciplinary approach knock-down mutant novel novel strategies overexpression phosphoric diester hydrolase public health relevance receptor respiratory smooth muscle response sensor time use tool

项目摘要

A large number of G protein coupled receptors (GPCR) utilize cAMP as their second messenger to induce alterations in cell function. In fact, in the same cell several different GPCR can increase cAMP, leading to the question of how the cell interprets the signals from these receptors differently. The concept of cAMP compartmentation, where the second messenger is not generated uniformly throughout the cell, is readily accepted yet poorly understood. We have found that the enzymes that synthesize cAMP, adenylyl cyclases (AC's), are not uniformly distributed through the plasma membrane. Furthermore, GPCR can preferentially couple to certain AC isoforms due to colocalization in lipid rafts. While we have made progress in understanding how specific receptors can couple to certain AC's, little progress has been made in defining the compartments of cAMP inside cells and even less is known about what cellular responses can be modified by different pools of cAMP. One problem has been that common cell models used in the field lack highly compartmentized cAMP pools. We have found that cultured human airway smooth muscle (HASM) cells express identifiable cAMP compartments. Furthermore, we can define these compartments based on the isoforms of AC they express. We have observed that signaling by cAMP generated by AC2, but not AC6 or other AC's, leads to the expression of IL-6 by HASM. Moreover, cAMP generated by AC6, but not by AC2, increases expression of somatostatin and stimulates a cell shape change called arborization. Thus, AC2- and AC6-specific responses can be used to define the cAMP signaling compartments in HASM. The goal of this project is to characterize the other components of these two cAMP compartments by using overexpression and knockdown of specific AKAP's and PDE's. Novel AC mutants will be used to manipulate AC localization and function to determine how these pools are assembled. This project proposes novel, multidisciplinary approaches to define the components responsible for establishing and maintaining cAMP signaling compartments. Results will have broad applicability due to the fundamental nature of cAMP signaling, but because a well-differentiated cell model is used, our findings will also have direct relevance to asthma and COPD therapy. GM107094 Ostrom, Rennolds S 2.

大量 G 蛋白偶联受体 (GPCR) 利用 cAMP 作为第二信使诱导细胞功能的改变。事实上，在同一个细胞中几种不同的GPCR都可以增加cAMP，导致细胞如何以不同的方式解释来自这些受体的信号的问题。这 cAMP 区室的概念，其中第二信使不是均匀生成的在整个细胞中，很容易被接受但了解甚少。我们发现酶合成 cAMP、腺苷酸环化酶 (AC)，在血浆中分布不均匀膜。此外，由于共定位，GPCR 可以优先与某些 AC 亚型偶联在脂筏中。虽然我们在理解特定受体如何耦合方面取得了进展对于某些 AC，在定义细胞内 cAMP 的区室方面进展甚微，并且关于不同的 cAMP 池可以改变哪些细胞反应，人们知之甚少。一问题是该领域使用的常见细胞模型缺乏高度分隔的 cAMP 库。我们发现培养的人气道平滑肌 (HASM) 细胞表达可识别的 cAMP 隔间。此外，我们可以根据 AC 的异构体来定义这些区室表达。我们观察到，AC2（而不是 AC6 或其他 AC）产生的 cAMP 信号会导致 HASM 表达 IL-6。此外，AC6 而非 AC2 产生的 cAMP 会增加生长抑素的表达并刺激称为树枝化的细胞形状变化。因此，AC2-和 AC6 特定响应可用于定义 HASM 中的 cAMP 信号区室。目标该项目的目的是通过使用来表征这两个 cAMP 隔室的其他组件特定 AKAP 和 PDE 的过度表达和敲低。新型 AC 突变体将用于操纵 AC 定位和功能来确定这些池的组装方式。这个项目提出新颖的、多学科的方法来定义负责建立的组成部分并维持 cAMP 信号传导室。结果将具有广泛的适用性，因为 cAMP 信号传导的基本性质，但由于使用了分化良好的细胞模型，我们的研究结果还将与哮喘和慢性阻塞性肺病的治疗直接相关。 GM107094 奥斯特罗姆，雷诺兹 S 2.