Functional and Mechanistic Dissection of GPCR Endosomal Signaling Dynamics

摘要 G蛋白偶联受体(GPCRs)是构成最大类药物靶点的关键信号。 翻译细胞外刺激以调节人类生理的蛋白质。因此，理解gpcr 信号级联将提供对这些复杂生理机制背后的分子机制的洞察 过程，它们在疾病中的扰动，并告知我们设计更有效的治疗方法的能力。在最近 多年来，我的实验室和其他人已经做出了一个改变范式的发现，即GPCRs可以在药物- 诱导从质膜内化到内体隔间。然而，有一点很重要 仍未探索的变量是GPCR内体cAMP活性的时间动态如何影响 下行信令。在目前的提案中，我将剖析GPCR激活持续时间的影响 内小体对下游信号的影响，识别调节受体时空的新蛋白质复合体 信号动力学，并将这一知识应用于与生理相关的系统--海马神经元。通过 结合功能基因组学、蛋白质组学和光遗传学，该提案旨在提供一种全面的 了解内体中gpr信号的时间动态如何调节生物功能。 这项工作对于理解GPCR信号转导机制和识别新的信号转导机制是必不可少的 途径，为药物治疗提供新的靶点。

Abstract G protein-coupled receptors (GPCRs), which compose the largest class of drug targets, are critical signaling proteins that translate extracellular stimuli to mediate human physiology. As such, understanding GPCR signaling cascades will provide insight into the molecular mechanisms underlying these complex physiological processes, their perturbation in disease, and inform our ability to design more efficient therapeutics. In recent years, my lab and others have made a paradigm-shifting discovery that GPCRs can be activated after drug- induced internalization from the plasma membrane into endosomal compartments. However, one essential variable that remains unexplored is how the temporal dynamics of GPCR endosomal cAMP activity impact downstream signaling. In the current proposal, I will dissect the influence of the duration of GPCR activation in endosomes on downstream signaling, identify novel protein complexes that regulate receptor spatiotemporal signaling dynamics, and apply this knowledge to a physiologically relevant system—hippocampal neurons. By combining functional genomics, proteomics, and optogenetics, this proposal aims to provide a comprehensive understanding of how the temporal dynamics of GPCR signaling in endosomes regulates biological functions. This work is essential to comprehending the mechanisms of GPCR signaling and identifying novel signaling pathways, providing new targets for pharmacologic therapies.