Heroin-Induced genomic regulation of Ventral Pallidum neuron subtypes

项目摘要/摘要 近年来，阿片类药物的使用、依赖和成瘾急剧增加，达到流行的比例， 导致巨大的经济和社会健康负担，以及越来越多的过量用药。至 抗击这一流行病，我们必须了解导致阿片类药物的神经生物学基础 使用无序。我们必须在阿片类药物使用障碍和失调中确定大脑中受损的神经元亚型 这些神经元中的分子构成了细胞、回路和最终行为适应的基础。使用RATE 药物自我给药(SA)和复发试验被认为是最好的可用动物模型 成瘾，将使我们能够更全面地了解基因组背后的分子机制， 表观遗传和转录诱导的细胞可塑性，驱动着持久的药物寻求和倾向 海洛因复吸者。 我们将进行全基因组转录组和开放染色质图谱的腹侧苍白球(VP)投射 大鼠海洛因依赖后急性停药和长期服药后的神经元亚型 禁欲。在这里，我们将重点放在VP作为大脑奖励回路中的一个关键节点。我们的研究将分析副总裁 投射到伏核、腹侧被盖区、丘脑内侧背侧和外侧的神经元 缰绳。然后我们将把转录和表观基因组数据与互补的转录和 表观基因组数据集，包括来自脑计划的描述VP细胞类型多样性的多模式数据 及其输出电路。我们将重建细胞类型特异的基因共表达并开放染色质网络 并确定Hub基因预计在立即和长期戒除海洛因方面发挥核心作用，这 可能是随后复发行为的基础。此数据集和模型集合将可用 通过一个生物学家友好的门户网站，基于我们的大脑倡议资助的神经科学多组学分析 站台。 利用产生的数据，我们将开发大鼠基因座特异性CRISPR表观组靶向工具来确定 VP投射神经元亚型调控的关键中枢基因的功能意义。要做到这一点 目标，我们将使用大鼠复发模型结合先进的CRISPRa和CRISPRi AAV工具来 在海洛因滥用或戒毒期间增强或减少关键HUB基因的转录 提示诱导复职。在这项拨款申请中建议的研究将首次确定 海洛因诱导的基因网络适应在时间上以细胞类型特异的方式发生在 新颖的神经生物回路。

PROJECT SUMMARY/ABSTRACT Opioid use, dependence, and addiction have dramatically increased to epidemic proportions in recent years, leading to substantial financial and societal health burdens, as well as an increasing number of overdoses. To combat this epidemic, it is imperative that we understand the neurobiological underpinnings that lead to opioid use disorder. We must identify disrupted neuron subtypes in the brain in opioid use disorders and dysregulated molecules within these neurons that underlie cellular, circuit, and ultimately behavioral adaptations. Use of rat drug self-administration (SA) and relapse assays, which are considered the best available animal models of addiction, will allow a more complete understanding of the molecular mechanisms underlying the genomic, epigenetic, and transcriptional-induced cellular plasticity that drives the long-lasting drug seeking and propensity for heroin relapse. We will perform genome-wide transcriptome and open-chromatin profiling of ventral pallidum (VP) projection neuron subtypes in rat heroin SA, both acutely following drug cessation and after prolonged periods of drug abstinence. Here, we focus on the VP as a critical node in the brain’s reward circuit. Our studies will profile VP neurons that project to the nucleus accumbens, ventral tegmental area, medial dorsal thalamus, and lateral habenula. We will then integrate the transcriptomic and epigenomic data with complementary transcriptomic and epigenomic datasets, including multimodal data from the BRAIN Initiative describing cell type diversity in the VP and its output circuits. We will reconstruct cell type-specific gene co-expression and open chromatin networks and identify hub genes predicted to have central roles in immediate and prolonged abstinence from heroin, which could underlie subsequent relapse behavior. This collection of datasets and models will be made available through a biologist-friendly web portal based on our BRAIN Initiative-funded Neuroscience Multi-Omic Analytics platform. Using the data generated we will develop rat gene loci-specific CRISPR epigenomic targeting tools to determine the functional significance of key hub genes that are regulated in VP projection neuron subtypes. To achieve this goal, we will employ rat models of relapse in combination with advanced CRISPRa and CRISPRi AAV tools to enhance or reduce transcription of key hub genes during heroin SA or abstinence from heroin SA followed by cue-induced reinstatement. The studies proposed in this grant application will, for the first time, identify the distinct heroin-induced gene network adaptations occurring temporally in a cell-type-specific manner within a novel neurobiological circuit.