Compartmentalized signaling and crosstalk in airway myocytes

项目总结/摘要 这项基础科学基金旨在确定生理相关的基本机制， 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。我们提出 基于我们多样化的调查团队最近的发现，描述了3种不同的GS偶联受体如何在 气道平滑肌（ASM）细胞--E2 AR、EP 2和EP 4受体--的信号是区室化的 在细胞内调节重要的ASM细胞功能。目标1将采用新的成像方法来描绘 每个受体cAMP/PKA信号传导的时空特征，并证明了这是如何划分的 信号传导是由AKAP和PDE同种型的受体特异性互补物以及竞争性共 激活的Gq偶联受体。AIM 1还将采用多个亚细胞靶向生物传感器来表征 每个受体从细胞内膜隔室发出信号的能力。目标2将评估这些 不同的受体产生独特的磷酸化蛋白质组签名，以及如何操纵机制， 形成局部cAMP/PKA信号调节这些特征。目标3将确定机制如何 决定ASM cAMP和ASM蛋白质组的时空特征影响Gs-coupled GPCR调控 ASM收缩、迁移和合成功能。拟议的研究将为以下方面奠定基础： 以方法进步和知识的形式理解划分信号 Gs偶联受体如何利用不同的信号传导机制， 功能效应。从翻译的角度来看，我们的发现将构成一个关键的基础科学基础 用于开发靶向信号室机制的新药，最容易应用于更好的 控制哮喘的特征，如气道高反应性、气道重塑， 炎症

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.