Molecular signal transduction of cAMP compartments

cAMP 区室的分子信号转导

• 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
• 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

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.

大量的G蛋白耦合受体（GPCR）将营地作为第二使者 诱导细胞功能改变。实际上，在同一细胞中，几种不同的GPCR可以增加营地， 导致了一个问题，即细胞如何以不同的方式解释这些受体的信号。这 营地隔间的概念，其中第二使者不是均匀产生的 在整个细胞中，很容易被接受但知之甚少。我们发现酶 合成cAMP，Adenylyl循环酶（AC）不均匀地通过等离子体分布 膜。此外，由于共定位，GPCR可以优先将其与某些AC同工型相结合 在脂质筏中。尽管我们在理解特定受体如何融入的过程中取得了进步 在定义牢房内部的营地和 关于哪些细胞反应可以通过不同的cAMP池可以修改哪些细胞反应的了解更少。一 问题是该领域使用的常见细胞模型缺乏高度分室的营地池。 我们发现，培养的人气道平滑肌（HASM）细胞表达可识别的cAMP 车厢。此外，我们可以根据AC的同工型来定义这些隔室 表达。 我们已经观察到由AC2产生的营地（而不是AC6或其他AC）发出的信号导致 HASM的IL-6表达。此外，AC6产生的营地，但不是AC2产生 生长抑素的表达并刺激称为树博化的细胞形状变化。因此，AC2-和 AC6特异性响应可用于定义HASM中的cAMP信号隔室。目标 该项目的特征是通过使用这两个营地的其他组件 特定AKAP和PDE的过表达和敲低。新颖的交流突变体将习惯 操纵AC定位和功能，以确定如何组装这些池。这个项目 提出了新颖的多学科方法来定义负责建立的组件 并维护营地信号隔间。由于 营地信号的基本性质，但由于使用了差异化的细胞模型，因此 发现也将与哮喘和COPD疗法直接相关。 GM107094 Ostrom，Rennolds S 2。