Discovery of novel pharmacotherapeutic targets for opioid addiction

发现阿片类药物成瘾的新药物治疗靶点

• 批准号: 10616803
• 负责人: Alexander Wanless Smith
• 金额: $ 24.9万
• 依托单位: MEDICAL UNIVERSITY OF SOUTH CAROLINA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10616803
• 关键词: Abstinence; Adherence; Affect; Agonist; Analgesics; Animals; Area; Atlases; Behavior; Big Data; Bioinformatics; Biological; Brain; Brain region; Cells; Clinical; Corpus striatum structure; Cues; Data; Data Analyses; Data Set; Detection; Development; Disease; Dopamine; Epidemic; Extinction; FDA approved; FOS gene; Fluorescence-Activated Cell Sorting; Gene Expression; Gene Targeting; Genes; Genetic; Genetic Recombination; Glutamates; Goals; Human; Immediate-Early Genes; Immunohistochemistry; Intake; Knowledge; Lipids; Mentors; Messenger RNA; Methods; Molecular; Molecular Biology; Mus; Narcotic Analgesics; Nature; Neuroanatomy; Neurobiology; Neurons; Opiate Addiction; Opioid; Opioid Receptor; Oxycodone; Persons; Pharmaceutical Preparations; Phase; Population; Prevention; Property; Proteins; Publishing; Pythons; Rattus; Relapse; Replacement Therapy; Reproducibility; Research; Research Personnel; Resources; Reverse Transcriptase Polymerase Chain Reaction; Self Administration; Site; Structure; System; Techniques; Testing; Therapeutic; Tissues; Training; Translating; Translational Research; United States; Vulnerable Populations; Western Blotting; abuse liability; addiction; brain circuitry; career; designer receptors exclusively activated by designer drugs; efficacy evaluation; experience; experimental study; large datasets; mouse model; neural circuit; neuroadaptation; neuropsychiatric disorder; non-opioid analgesic; novel; novel therapeutics; opioid abuse; opioid epidemic; opioid overdose; pharmacologic; programs; protein expression; relapse prevention; response; transcriptome sequencing; transcriptomics; translational potential

PROJECT SUMMARY/ABSTRACT The US is in the midst of an opioid abuse and overdose epidemic. Oxycodone is one of the most prescribed analgesics, is the first opioid many people experience, and has physiochemical properties that allow it to accumulate in the brain at rates higher than other opioids, perhaps explaining its considerable abuse potential. Here, I seek to perform high-throughput experiments to generate brain-wide data on the cellular and molecular mechanisms of cue-induced reinstatement to oxycodone seeking. Specifically, I propose to use FosCreER mice to fluorescently `tag' neuronal ensembles activated by cue-induced reinstatement. I will then use cutting-edge transcriptomics to identify relapse-related genes in these cellular ensembles that drive reinstatement. I will then prioritize, and test, whether these genes may serve substrates for the development of novel medications to prevent relapse using a ”circuit therapeutic” approach. In aim 1, iDISCO+, a lipid clearing method, will produce brain-wide data on regions activated by relapse. I will then use ClearMap, a published Python package, to conduct high-throughput detection of activated neurons and registration of coordinates onto the Allen Brain Atlas. I will then rigorously examine this large data set, and test whether reinstatement-responsive cell ensembles in prioritized structures contribute to relapse behavior. In aim 2, I will focus on cellular populations in brain regions shown to be required for relapse-related behavior, and will again tag neurons activated by reinstatement in these sites. Tissue will be dissected, and fluorescence activated cell sorting will be used to isolate these activated neurons for trancriptomic profiling via RNA-Seq. Together, these two aims will yield large data sets directly related to the neurocircuitry and molecular biology of reinstatement of oxycodone seeking. I will once again rigorously analyze these data sets, with strict adherence to pre-established criteria, to prioritize reinstatement-responsive genes most likely to represent efficacious targets for development of novel pharmacotherapeutics. Following completion of the training phase, in aim 3 I will extend these findings to both rat and mouse models of self-administration, and will validate that these transcriptomic adaptations occur across species, and that they are detected as changes in functional proteins. Once again, I will rigorously prioritize these targets for translational potential. In aim 4 I will use pharmacological agents that modulate prioritized protein targets to determine whether they can block cue-induced reinstatement of oxycodone seeking, focusing on compounds that may be able to quickly move into clinical settings. The training added in this proposal will allow me to emerge as an exceptionally well-rounded independent investigator. During my independent career I will perform translational research to develop novel circuit-based therapeutics for prevention of relapse.

项目概要/摘要 美国正处于阿片类药物滥用和过量流行之中。羟考酮是最常用的药物之一 镇痛药，是许多人使用的第一种阿片类药物，具有使其能够 在大脑中的积累速度高于其他阿片类药物，这或许可以解释其巨大的滥用潜力。 在这里，我寻求进行高通量实验来生成有关细胞和分子的全脑数据 线索诱导恢复羟考酮寻求的机制。具体来说，我建议使用 FosCreER 小鼠 荧光“标记”由提示诱导恢复激活的神经元群。然后我会使用最先进的 转录组学来识别这些驱动康复的细胞群中与复发相关的基因。然后我会 优先考虑并测试这些基因是否可以作为开发新药物的底物 使用“循环治疗”方法预防复发。在目标 1 中，iDISCO+（一种脂质清除方法）将产生 有关复发激活区域的全脑数据。然后我将使用 ClearMap（一个已发布的 Python 包）来 对激活的神经元进行高通量检测并将坐标注册到艾伦大脑上 阿特拉斯.然后我将严格检查这个大数据集，并测试恢复响应细胞是否 优先结构中的集合会导致复发行为。在目标 2 中，我将重点关注细胞群体 在被证明是复发相关行为所需的大脑区域中，并将再次标记由 在这些站点中恢复。将解剖组织，并使用荧光激活细胞分选 通过 RNA 测序分离这些激活的神经元进行转录组分析。这两个目标结合在一起将产生 与恢复羟考酮的神经回路和分子生物学直接相关的大数据集 寻求。我将再次严格分析这些数据集，严格遵守预先制定的标准， 优先考虑最有可能代表发展的有效目标的恢复反应基因 新的药物治疗。培训阶段完成后，在目标 3 中，我将把这些发现扩展到 大鼠和小鼠的自我给药模型，并将验证这些转录组适应的发生 跨物种，并且它们被检测为功能蛋白的变化。我将再次严格要求 优先考虑这些目标的转化潜力。在目标 4 中，我将使用调节药物 确定蛋白质目标的优先顺序，以确定它们是否可以阻止提示诱导的羟考酮恢复 寻找、专注于可能能够快速进入临床环境的化合物。培训内容添加到 这项建议将使我成为一名非常全面的独立调查员。在我的 独立的职业生涯我将进行转化研究，开发新型的基于回路的疗法 预防复发。