Molecular basis for ligand and cell type specific regulation of opioid receptors

摘要/项目摘要。 阿片类药物滥用和成瘾是主要的公共卫生问题。阿片类药物是非常有用的止痛药，但 据估计，美国有200万人患有与滥用处方阿片类药物有关的疾病。这个 这些疾病造成的社会和经济代价是毁灭性的，而且还在上升：每年平均有44人死亡 处方阿片类药物过量的一天。阿片类药物是结构多样的分子，包括羟考酮， 海洛因和内啡肽，以及这些分子的生理效应是由G蛋白偶联介导的 受体(GPCRs)。最近开发的“有偏见的”阿片类激动剂表明，这种多样性可以被挖掘出来 确定副作用较小的药物，但这些“有偏见的”激动剂的作用机制仍然存在 没有完全解决。现在，新的技术进步使生化捕获蛋白质成为可能。 相互作用网络以亚分钟的时间分辨率从活细胞内部介导GPCR活性。 捕获、量化和表征调节阿片类药物的内源性蛋白质的能力 活性为确定偏向激动剂与内源性配体或滥用药物有何不同打开了新的大门。 这一K99/R00奖项将尖端蛋白质组学方面的关键新培训与传统细胞生物学相结合 检查偏向阿片激动剂机制的技术：目标1--定义不同的动力学 阿片类药物改变了u阿片受体的位置及其与其转导和调节的偶联 蛋白质；目标2--确定受标准或有偏见的阿片类药物刺激的新的蛋白质调节因子。目标3- 定义µOR信号靶点，并确定这些蛋白质在阿片类药物之间是否存在差异。基于以下方面的未来研究 这些结果将有助于确定阿片受体如何在正常、药物激活或 由于滥用药物和上瘾而导致的疾病状态。

Abstract/Project Summary. Opioid abuse and addiction are major public health concerns. Opioids are highly useful analgesics, yet an estimated two million people in the United States suffer disorders related to abuse of prescription opioids. The social and economic costs of these disorders are devastating and on the rise: an average of 44 people die every day from prescription opioid overdoses. Opioids are structurally diverse molecules that include oxycodone, heroin and endorphins, and the physiological effects of these molecules are mediated by G protein-coupled receptors (GPCRs). Recently developed `biased' opioid agonists demonstrate that this diversity can be mined to identify drugs with less harmful side effects, but the mechanism of action of these `biased' agonists remains incompletely resolved. New technical advances now make it possible to biochemically capture the protein interaction networks mediating GPCR activity from inside of living cells with sub-minute temporal resolution. The ability to capture, quantify, and characterize the endogenous proteins which mediate and regulate opioid activity opens new doors for determining how biased agonists differ from endogenous ligands or drugs of abuse. This K99/R00 award combines critical new training in cutting-edge proteomics with traditional cell biological techniques to examine the mechanism of biased opioid agonism: Aim 1-Define the kinetics by which different classes of opioids alter mu opioid receptor (µOR) location and coupling to its transducer and regulatory proteins; Aim 2-Identify new protein regulators of µOR stimulated by standard or biased opioids. Aim 3- Define µOR signaling targets and determine if these proteins differ between opioids. Future studies based on these results will help to define how opioid receptors operate under normal, pharmacologically activated, or disease states resulting from drug abuse and addiction.