Proteomics/Genomics of Opiate Analgesia and Addiction

药物描述（申请人提供）：吗啡是一种高效的阿片类镇痛药，在现代医学中是一种有价值的止痛药。然而，长期吗啡给药的缺点是成瘾性的发展与耐受性和依赖性。理想地，设计用于有效镇痛的治疗剂将是有利的，其缺乏吗啡和相关阿片类药物的成瘾倾向。考虑到这一目标，有必要了解介导吗啡诱导镇痛的生物化学途径与耐受性和依赖性相比。当新的创新技术被应用于阐明具体的研究问题时，往往会出现科学进步。因此，这项R21的试点应用，创新的研究，利用国家的最先进的技术在蛋白质组学和基因组学，将阐明参与吗啡诱导的镇痛相比，耐受性或依赖性的分子组成部分。值得注意的是，β-抑制蛋白基因敲除小鼠在吗啡诱导的作用中显示出明显的变化，对应于增强的镇痛、缺乏耐受性和依赖性没有变化。这些发现导致不同的生物化学途径的这三个参数的吗啡行动的假设。因此，该可行性项目的目标将是利用蛋白质组学和基因组学来比较慢性吗啡治疗期间β-arrestin敲除小鼠和野生型小鼠中受调节的分子组分，作为阐明与耐受性或依赖性相比吗啡诱导的镇痛的不同生化途径的手段。具体目的是（1）与野生型小鼠相比，β-抑制蛋白敲除小鼠脑中慢性吗啡期间受调节的蛋白质和肽的蛋白质组学评价，以及（2）在β-抑制蛋白敲除小鼠和野生型小鼠中使用独特的神经特异性小鼠cDNA微阵列对慢性吗啡期间差异基因表达的基因组学评价。这些研究将利用最先进的设施，蛋白质组学和质谱，基因组学与神经特异性小鼠cDNA微阵列。将分析调控基因产物与基因家族的一级序列同源性，以预测其生物学功能。在β-arrestin基因敲除和野生型小鼠的调控基因产物的比较将允许分配的调控蛋白和基因的候选人参与吗啡镇痛，耐受性，或依赖的生化途径。阐明不同的途径将是重要的，因为它可以允许未来设计的阿片类药物治疗方案，提供有效的镇痛，没有成瘾的耐受性和依赖性。

DESCRIPTION (provided by applicant): Morphine is a highly effective opiate analgesic, and is a valuable painkiller in modern medicine. However, the disadvantage of chronic morphine administration is the development of addiction with tolerance and dependence. Ideally, it would be advantageous to design therapeutics for effective analgesia, which lacks the addictive liabilities of morphine and related opiates. With this goal in mind, it is necessary to understand the biochemical pathways that mediate morphine-induced analgesia compared to tolerance, and dependence. Scientific advances often occur when new innovative technologies are applied towards elucidating specific research issues. Therefore, this R21 application for pilot, innovative studies utilizing state-of-the-art technologies in both proteomics and genomics, will elucidate molecular components involved in morphine-induced analgesia compared to tolerance or dependence. Notably, beta-arrestin knockout mice show distinct changes in morphine-induced actions corresponding to enhanced analgesia, absence of tolerance, and no change in dependence. These findings lead to the hypothesis of distinct biochemical pathways for these three parameters of morphine actions. Therefore, the goal of this feasibility project will be to utilize both proteomics and genomics to compare regulated molecular components in beta-arrestin knockout mice and wild-type mice during chronic morphine treatment, as a means to elucidate distinct biochemical pathways for morphine-induced analgesia compared to tolerance or dependence. The specific aims are (1) proteomic evaluation of regulated proteins and peptides during chronic morphine in brains of beta-arrestin knockout mice compared to wild-type mice, and (2) genomic evaluation of differential gene expression during chronic morphine using unique neural-specific mouse cDNA microarrays in beta-arrestin knockout and wild-type mice. These studies will utilize state-of-the-art facilities for proteomics and mass spectrometry, and genomics with neural-specific mouse cDNA microarrays. Regulated gene products will be analyzed for primary sequence homology to gene families to predict their biological functions. Comparison of regulated gene products in beta-arrestin knockout and wild-type mice will allow assignment of regulated proteins and genes to candidate biochemical pathways involved in morphine analgesia, tolerance, or dependence. Elucidation of distinct pathways will be significant, since it could allow future design of opiate drug regimens that provide effective analgesia, without the tolerance and dependence of addiction.