Endosomal Platforms for Neuropeptide Receptor Signaling

神经肽受体信号转导的内体平台

• 批准号: 10093292
• 负责人: NIGEL W BUNNETT
• 金额: $ 28.3万
• 依托单位: NEW YORK UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-15 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10093292
• 关键词: Endosomal; Platforms; Neuropeptide; Receptor; Signaling

PROJECT SUMMARY This proposal examines the mechanisms by which G protein-coupled receptors (GPCRs) signal pain. Chronic pain is a hallmark of disease, a side effect of therapy, and a major cause of suffering. Although GPCRs mediate all aspects of nociception and are major therapeutic targets, the mechanisms by which GPCRs signal sustained pain are poorly understood, and clinical trials of GPCR antagonists in chronic pain often fail for unexplained reasons. The proposal challenges three dogmas that contribute to this lack of understanding: 1. GPCRs signal only from the cell-surface. 2. Endosomes are merely a conduit for GPCR recycling or degradation. 3. Cell-surface GPCRs are the optimal therapeutic target. The proposal hypothesizes that: 1. Endosomal GPCRs generate sustained signals that mediate persistent excitation of spinal neurons and nociception. 2. Targeting endosomal rather than cell-surface GPCRs is the ideal therapeutic strategy, and the clinical failure of conventional antagonists relates to their inability to inhibit endosomal receptors. Experiments will focus on substance P and calcitonin gene-related peptide receptors, which mediate central pain transmission and are internalized after painful stimuli. The contribution of receptor endocytosis to nociception will be evaluated using pharmacological and genetic approaches to disrupt clathrin, dynamin and β-arrestin, and by studying transgenic mice expressing non-internalizing receptors. Lipid-conjugation and nanoparticle- encapsulation will be used to deliver antagonists to endosomal GPCRs. Aim 1 will determine the contribution of endocytosis to somatic and colonic nociception in conscious mice. Aim 2 will define the importance of endocytosis for excitation of spinal neurons, which will be analyzed in intact tissues using electrophysiology. Aim 3 will determine the requirement of endocytosis for the generation of signals in subcellular compartments that underlie neuronal excitation and nociception, which will be studied in isolated neurons using biophysical, imaging and proteomic approaches. The results will provide fundamental information about pain signaling and therapy. Since GPCRs are the largest class of signaling proteins and the target of one half of therapeutic drugs, the outcomes will be broadly significant.

项目摘要 该建议检查G蛋白偶联受体（GPCRS）信号疼痛的机制。慢性的 虽然GPCR 介导伤害感受的所有方面，是主要的治疗靶标，GPCR信号的机制 持续的疼痛知之甚少，慢性疼痛中GPCR拮抗剂的临床试验常常失败 无法解释的原因。该提议挑战了三个缺乏理解的教条：1。 GPCR仅来自细胞表面的信号。 2。内体只是GPCR回收或 降解。 3。细胞表面GPCR是最佳的治疗靶点。该提议假设：1。 内体GPCR产生持续信号，介导脊柱神经元的持续兴奋和 伤害感受。 2。靶向内体而不是细胞表面GPCR是理想的疗法策略，并且 常规拮抗剂的临床失败与其无能抑制内体受体有关。实验 将专注于PESTANCE P和降钙素基因相关的肽受体，这些受体介导中心疼痛 传播并在疼痛刺激后内部化。接收器内吞作用对伤害感受的贡献 将使用药物和遗传学方法评估，以破坏网状蛋白，Dynamin和β-arrestin， 并通过研究表达非内在化受体的转基因小鼠。脂质结合和纳米颗粒 封装将用于向内体GPCR提供拮抗剂。 AIM 1将确定 有意识小鼠中的体细胞和结肠肿瘤的内吞作用。 AIM 2将定义的重要性 内吞作用可激发脊柱神经元，该神经元将在完整的组织中使用电生理学分析。 AIM 3将确定内吞作用对亚细胞隔室中信号产生的需求 神经元的兴奋和诺神经元素的基础，它将使用生物物理，在分离的神经元中研究 成像和蛋白质组学方法。结果将提供有关疼痛信号的基本信息 和治疗。由于GPCR是最大的信号蛋白类，也是一半的目标 治疗药物，结果将非常重要。