Compartmentalized signaling and crosstalk in airway myocytes

气道肌细胞中的区室化信号传导和串扰

基本信息

批准号：
10718208
负责人：
RENNOLDS S OSTROM
金额：
$ 58.16万
依托单位：
UNIVERSITY OF SOUTH ALABAMA
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-07-01 至 2027-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10718208
关键词：
A kinase anchoring protein Adenylate Cyclase Adult Affect Agonist Arrestins Asthma Basic Science Biology Biosensor Cell Nucleus Cell Separation Cell membrane Cell physiology Cells Characteristics Child Chronic Disease Clinical Complement Coughing Coupled Cyclic AMP Cyclic AMP-Dependent Protein Kinases Cytosol Data Disparate Dissociation EP4 receptor Elements Exhibits Foundations G Protein-Coupled Receptor Signaling G protein coupled receptor kinase G-Protein-Coupled Receptors GTP-Binding Proteins Goals Grant Heterotrimeric GTP-Binding Proteins Human Image Imaging Techniques Integral Membrane Protein Intracellular Membranes Kinetics Knowledge Link Location Measures Mediating Membrane Microdomains Methodology Molecular Biology Muscle Cells Muscle Contraction Muscle function Outcome Pathway interactions Phospholipase C Phosphoproteins Phosphorylation Phosphotransferases Physiological Post-Translational Protein Processing Protein Isoforms Proteins Proteome Receptor Signaling Regulation Relaxation Role Second Messenger Systems Shapes Shortness of Breath Signal Pathway Signal Transduction Smooth Muscle Myocytes Specificity Symptoms System Testing Visualization Wheezing Work airway hyperresponsiveness airway inflammation airway remodeling asthmatic design experimental study genetic regulatory protein grasp imaging approach insight knock-down mathematical model migration new therapeutic target novel overexpression preservation protein protein interaction receptor respiratory smooth muscle response spatiotemporal stoichiometry subcellular targeting tool

项目摘要

PROJECT SUMMARY/ABSTRACT This basic science grant seeks to determine fundamental mechanisms as to how a physiologically relevant cell controls the subcellular location of a signal to optimize the effect of that signal on cell function. We propose to build on recent findings by our diverse investigative team to delineate how 3 different Gs-coupled receptors in airway smooth muscle (ASM) cells - the 2AR, EP2, and EP4 receptors- have their signals compartmentalized within the cell to regulate important ASM cell functions. Aim 1 will employ novel imaging approaches to delineate spatiotemporal features of cAMP/PKA signaling by each receptor and demonstrate how this compartmentalized signaling is shaped by receptor-specific complements of AKAP and PDE isoforms, and by the competing co- activated Gq-coupled receptor. Aim 1 will also employ multiple subcellular-targeted biosensors to characterize the capacity of each receptor to signal from intracellular membrane compartments. Aim 2 will assess how these different receptors generate unique phosphoproteome signatures, and how manipulating the mechanisms shaping localized cAMP/PKA signaling regulates these signatures. Aim 3 will establish how the mechanisms dictating spatiotemporal features of ASM cAMP and the ASM proteome affect Gs-coupled GPCR regulation of ASM contraction, migration, and synthetic functions. The proposed studies will provide a foundation for understanding compartmentalized signaling in the form of both methodological advances and the knowledge gained in how Gs-coupled receptors employ distinct signaling mechanisms to render efficient and specific functional effects. From a translational perspective, our findings will constitute a critical basic science foundation for developing new drugs that target mechanisms of signaling compartments, most readily applied to better control asthma features such as airway hyperresponsiveness, airway remodeling, and possibly airway inflammation.

项目摘要/摘要这项基础科学赠款旨在确定有关身体相关的基本机制细胞控制信号的亚细胞位置，以优化该信号对细胞功能的影响。我们建议为了基于我们多样化的调查团队的最新发现，以描绘3种不同的GS耦合受体气道平滑肌（ASM）细胞 - 2AR，EP2和EP4受体 - 具有隔室的信号在细胞内调节重要的ASM细胞功能。 AIM 1将采用新颖的成像方法来描述每个接收器的CAMP/PKA信号传导的时空特征，并演示该分隔如何信号传导是通过AKAP和PDE同工型的接收器特异性完成的，以及竞争的共同体活化的GQ耦合受体。 AIM 1还将采用多个细胞靶向生物传感器来表征每个接收器从细胞内膜室信号的容量。 AIM 2将评估这些不同的受体产生独特的磷光体特征，以及如何操纵机制塑造局部营地/PKA信号传导调节这些签名。 AIM 3将确定机制如何指示ASM CAMP的时空特征和ASM蛋白质组影响GS耦合的GPCR调节 ASM收缩，迁移和合成功能。拟议的研究将为以方法学进步和知识的形式理解分隔的信号传导在GS耦合受体员工如何不同的信号传导机制方面获得了有效和特定的功能效应。从翻译的角度来看，我们的发现将构成关键的基础科学基金会用于开发针对信号隔室机制的新药，最容易应用于更好控制哮喘特征，例如气道高反应性，气道重塑和可能的气道炎。