Heroin-Induced genomic regulation of Ventral Pallidum neuron subtypes

海洛因诱导的腹侧苍白球神经元亚型的基因组调控

• 批准号: 10404982
• 负责人: Seth Abrams Ament
• 金额: $ 59.33万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 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 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10404982
• 关键词: ATAC-seq; Abstinence; Acute; Animal Model; Applications Grants; BRAIN initiative; Back; Behavior; Behavioral; Biological Assay; Brain; CRISPR interference; CRISPR-mediated transcriptional activation; Cell Nucleus; Cells; Chromatin; Clustered Regularly Interspaced Short Palindromic Repeats; Collection; Cues; Data; Data Set; Dependence; Dependovirus; Dorsal; Drug Addiction; Epidemic; Epigenetic Process; Foundations; Frequencies; Functional disorder; Funding; Gene Expression; Genes; Genetic; Genetic Transcription; Genomics; Globus Pallidus; Goals; Habenula; Health; Heroin; Heroin Dependence; Hour; Individual; Knowledge; Label; Lateral; Lead; Life; Link; Medial; Mediating; Messenger RNA; Modeling; Molecular; Neurobiology; Neurons; Neurosciences; Nucleus Accumbens; Opiate Addiction; Opioid; Output; Overdose; Pharmaceutical Preparations; Process; Proteins; Rattus; Regulation; Relapse; Research; Research Personnel; Rewards; Role; Self Administration; Thalamic structure; Therapeutic; Time; Tissues; Training; Transcription Process; Ventral Tegmental Area; addiction; cell type; chromatin immunoprecipitation; combat; dopaminergic neuron; drug abstinence; drug craving; drug seeking behavior; epigenome; epigenomics; falls; gene network; gene regulatory network; genome-wide; genomic locus; genomic tools; insight; molecular subtypes; multimodal data; multiple omics; nerve supply; network models; novel; opioid abuse; opioid use; opioid use disorder; psychostimulant; tool; transcriptome; transcriptome sequencing; transcriptomics; web portal

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.

项目总结/摘要 近年来，阿片类药物的使用、依赖和成瘾已急剧增加到流行病的程度， 导致巨大的财政和社会健康负担，以及越来越多的药物过量。到 与这种流行病作斗争，我们必须了解导致阿片类药物的神经生物学基础。 使用障碍。我们必须确定阿片类药物使用障碍和失调的大脑中被破坏的神经元亚型， 这些神经元内的分子是细胞、电路和最终行为适应的基础。使用大鼠 药物自我给药（SA）和复发试验，这被认为是最好的可用动物模型， 成瘾，将允许更完整地了解基因组， 表观遗传和转录诱导的细胞可塑性，驱动长期的药物寻求和倾向 因为海洛因复吸 我们将对腹侧苍白球（VP）投射进行全基因组转录组和开放染色质分析 大鼠海洛因SA中的神经元亚型，无论是急性停药后还是长期用药后 禁欲在这里，我们专注于VP作为大脑奖励回路中的关键节点。我们的研究将描绘副总统 投射到丘脑背侧核、腹侧被盖区、丘脑背内侧核和外侧核的神经元 缰然后，我们将整合转录组和表观基因组数据与互补的转录组和表观基因组数据。 表观基因组数据集，包括来自BRAIN Initiative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性基因共表达和开放染色质网络 并确定预测在立即和长期戒除海洛因中起核心作用的枢纽基因， 可能会导致随后的复发行为这些数据集和模型将在 通过一个生物学家友好的门户网站，基于我们的大脑计划资助的神经科学多细胞分析 平台 利用产生的数据，我们将开发大鼠基因座特异性CRISPR表观基因组靶向工具，以确定 VP投射神经元亚型中调控的关键中枢基因的功能意义。实现这一 为了实现这一目标，我们将采用大鼠复发模型与先进的CRISPRa和CRISPRi AAV工具相结合， 增强或减少海洛因SA或海洛因SA戒断期间关键枢纽基因的转录， 线索诱导的复职这项拨款申请中提出的研究将首次确定 不同的海洛因诱导的基因网络适应发生在一个细胞类型特异性的方式在时间上， 新的神经生物学回路