Precision pharmacology of the opioids

阿片类药物的精准药理学

• 批准号: 10242770
• 负责人: Christina Woo
• 金额: $ 50.7万
• 依托单位: HARVARD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-30 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10242770
• 关键词: Acetylation; Award; Behavior; Binding; Binding Sites; Biological; Biology; Cell surface; Cells; Chemicals; Data; Data Science; Euphoria; Exposure to; Gene Expression; Genetic; Genetic Transcription; Genome; Goals; Heroin; Internet; Maps; Mass Spectrum Analysis; Mediating; Metabolism; Modification; Molecular; Molecular Target; Morphine; Nature; Neurons; Non-Steroidal Anti-Inflammatory Agents; Nucleosomes; Opioid; Opioid Peptide; Outcome; Pathway interactions; Permeability; Pharmacology; Proteome; Regulation; Research; Site; Substance abuse problem; Technology; Time; addiction; base; epigenetic regulation; experimental study; insight; interest; mouse model; mu opioid receptors; new therapeutic target; novel; novel diagnostics; novel strategies; programs; protein complex; side effect; small molecule; structural biology; tissue culture

PROJECT SUMMARY/ABSTRACT Substance abuse behaviors result from molecular changes to gene expression programs in neurons over time. Morphine and heroin are highly addictive opioids that primarily stimulate the mu opioid receptor resulting in short- term euphoria. The addictive nature of morphine and heroin, unlike natural opioid peptides, may result from their cell permeability and thus direct influence on gene expression programs leading to addiction. However, understanding these precise molecular mechanisms is typically challenging due to the dearth of technologies to precisely map global molecular targets and pathways of a small molecule. We have developed an interdisciplinary precision pharmacology strategy to map the direct and indirect effects of a small molecule in the cellular proteome that we propose to apply to these addictive opioids. Our approach draws on the fields of chemical biology, mass spectrometry and data science to enable insight to the full range of molecular interactions, the structural biology underlying the interaction site, and changes to downstream pathways in a single experiment. The strategy involves: (1) treatment of whole cells with the small molecule of interest, (2) isolation of the resulting global molecular binding sites and (3) confident mass spectrometry-based assignment. We recently applied this platform to study three non-steroidal anti-inflammatory drugs (NSAIDs). Our results revealed several protein complexes involved in gene expression that the NSAIDs interact with, including a directly with the nucleosome. These results point to the vast web of molecular mechanisms that is now observable by precision pharmacology. In this proposal, we will apply our technology to morphine and heroin to determine their direct influence on gene expression leading to substance abuse. We will first develop a set of “click opioids” and “photo-click opioids” as generally useful probes for tracking opioid mechanisms in biology. We will specifically apply these probes to characterize opioid-driven genetic and epigenetic regulation in neuronal cells first in tissue culture and subsequently to mouse models of addiction. With a global map of the opioid interactome, these data will reveal direct opioid interactions with nucleosomes by binding, covalent modification, or mediating acetylation marks, and the indirect influence on upstream transcriptional programs that drive gene expression changes. By characterizing the broader interactions of the opioids, we are poised to expose molecular mechanisms leading to addiction, identify novel targets for new therapeutics and diagnostics, and open new paradigms in biological regulation by small molecules. The outlined research is a conceptually novel approach to perform mechanism of action studies that is suitable for the Avenir Award due to the broad potential for impact and early stage of this research.

项目概要/摘要 药物滥用行为是由于神经元基因表达程序随着时间的推移发生分子变化而导致的。 吗啡和海洛因是高度成瘾的阿片类药物，主要刺激μ阿片受体，导致短暂的 术语欣快感。与天然阿片肽不同，吗啡和海洛因的成瘾性可能是由于它们的 细胞通透性从而直接影响导致成瘾的基因表达程序。然而， 由于缺乏技术，理解这些精确的分子机制通常具有挑战性。 精确绘制小分子的全局分子靶点和途径。我们开发了一个 跨学科精准药理学策略，绘制小分子在药物中的直接和间接影响 我们建议将细胞蛋白质组应用于这些成瘾的阿片类药物。我们的方法借鉴了以下领域 化学生物学、质谱和数据科学，使人们能够深入了解分子的全部范围 相互作用、相互作用位点背后的结构生物学以及下游途径的变化 单一实验。该策略包括：(1) 用感兴趣的小分子处理整个细胞，(2) 分离所得的全局分子结合位点和（3）基于可信质谱的分配。 我们最近应用这个平台来研究三种非甾体抗炎药（NSAID）。我们的成果 揭示了几种与非甾体抗炎药相互作用的基因表达相关的蛋白质复合物，包括 直接与核小体结合。这些结果指出了目前存在的庞大的分子机制网络 可通过精密药理学观察。在这个提案中，我们将把我们的技术应用于吗啡和海洛因 确定它们对导致药物滥用的基因表达的直接影响。我们首先会开发一套 “点击阿片类药物”和“照片点击阿片类药物”通常是追踪生物学中阿片类药物机制的有用探针。 我们将专门应用这些探针来表征神经元中阿片类药物驱动的遗传和表观遗传调控 细胞首先进入组织培养物，随后进入成瘾小鼠模型。拥有阿片类药物的全球地图 相互作用组，这些数据将揭示阿片类药物通过结合、共价修饰与核小体的直接相互作用， 或介导乙酰化标记，以及对驱动基因的上游转录程序的间接影响 表情发生变化。通过描述阿片类药物更广泛的相互作用，我们准备揭露 导致成瘾的分子机制，确定新疗法和诊断的新靶标，以及 开启小分子生物调控的新范式。概述的研究在概念上是新颖的 由于具有广泛的潜力，适合 Avenir 奖的行动机制研究方法 了解这项研究的影响和早期阶段。