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镇痛剂，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。的 这些疾病造成的社会和经济损失是毁灭性的，而且还在上升：平均每年有44人死亡。 因处方阿片类药物过量死亡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，这些分子的生理作用是由G蛋白偶联的 受体（GPCR）。最近开发的“偏向性”阿片激动剂表明，这种多样性可以被挖掘， 确定副作用较小的药物，但这些“偏向性”激动剂的作用机制仍然存在 不完全解决。新的技术进步现在使生物化学捕获蛋白质成为可能 从活细胞内部以亚分钟时间分辨率介导GPCR活性的相互作用网络。 捕获、定量和表征介导和调节阿片类药物的内源性蛋白质的能力 活性为确定偏向性激动剂与内源性配体或滥用药物的区别打开了新的大门。 这个K99/R 00奖项将尖端蛋白质组学的关键新培训与传统的细胞生物学相结合， 技术来检查偏向阿片激动机制：目的1-定义不同的动力学 阿片类药物的种类改变μ阿片受体（μOR）的位置及其与其转导子和调节子的偶联， 目标2-识别由标准或偏倚阿片类药物刺激的µOR的新蛋白质调节剂。目标3- 定义µOR信号靶标，并确定这些蛋白质在阿片类药物之间是否存在差异。未来研究基于 这些结果将有助于确定阿片受体如何在正常，非激活，或 由于药物滥用和成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.