Regulation of Gene Enhancers in Human Heroin Use

项目摘要/摘要--项目4 S计划的目标是表征基因组增强子区的影响，这些增强子区能够发挥作用 在鸦片成瘾中，对基因表达的关键控制。在几个组织中的研究表明 增强子区域被几个特定的组蛋白修饰标记，例如H3K27ac(组蛋白3 Lys27 乙酰化)，并富含几种染色质重塑因子。一个例子是BRD4，它是 BET(溴结构域和末端外)亚家族，通过其 溴域，并与几个组织中的增强剂紧密相连。我们发现了组织水平升高的 海洛因使用障碍患者纹状体内H3K27ac和BRD4的表达 强烈地与多年的海洛因使用和关键基因网络的表达水平有关。类似的监管也出现在 自我给药海洛因的大鼠的纹状体。此外，我们还发现，破坏物质的小分子 BRD4/BET功能-在癌症治疗的临床开发中-在海洛因中发挥治疗样效应 啮齿类动物的成瘾模型。该项目将在几个方面促进这些研究。我们将获得基因组- 海洛因使用障碍患者和配对对照组NAC和DS中增强子区域的广泛图谱， 而在海洛因自我给药后的大鼠中，使用CHIP-SEQ对H3K27ac和BRD4等标记进行检测，结果表明， 与基因和染色质分析核心相结合。这将在分离的核上单独执行 来自这些脑区的神经元和非神经元细胞。我们将与Core合作开发高级 覆盖这些数据集的生物信息学方法，这是可靠地识别增强剂所必需的， 以及我们目前在相同样本上生成的RNA-seq和atac-seq数据集，以及 将进一步深入了解受已确定的海洛因调节增强剂影响的特定靶基因。 我们将通过研究双向作用来检验增强子调控在海洛因成瘾中的因果作用。 NAC和DS中BRD4对大鼠模型基因表达和行为终点的影响，包括 筛选几种BRD4/BET抑制剂的抗成瘾作用。这些研究提供了一种翻译 有机会推进治疗药物成瘾的新疗法。

PROJECT SUMMARY/ABSTRACT– PROJECT 4 Project 4’s objective is to characterize the influence of enhancer regions of the genome, which exert crucial control over gene expression, in opiate addiction. Work in several tissues has demonstrated that enhancer regions are “marked” by several specific histone modifications, such as H3K27ac (histone 3 Lys27 acetylation), and are enriched in several chromatin remodeling factors. One example is BRD4, a member of the BET (bromodomain and extra-terminal) subfamily, which binds acetylated histones through its bromodomains and is tightly linked with enhancers in several tissues. We have found elevated tissue levels of both H3K27ac and BRD4 in striatum of humans with heroin use disorders, with such elevations correlating strongly with years of heroin use and expression levels of key gene networks. Similar regulation is seen in striatum of rats that self-administer heroin. Moreover, we have found that small molecules that disrupt BRD4/BET function—in clinical development for cancer treatment—exert therapeutic-like effects in heroin addiction models in rodents. This Project will further these studies in several ways. We will obtain genome- wide maps of enhancer regions in NAc and DS of humans with heroin use disorders and matched controls, and in rats after heroin self-administration, by use of ChIP-seq for H3K27ac and BRD4, among other marks, in conjunction with the Gene and Chromatin Analysis Core. This will be performed separately on nuclei isolated from neurons and non-neuronal cells of these brain regions. We will work with the Core on advanced bioinformatics approaches to overlay these datasets, which is required for reliable identification of enhancers, along with RNA-seq and ATAC-seq datasets that we are currently generating on the same samples and that will provide further insight into the specific target genes affected by the identified heroin-regulated enhancers. We will test the causal role of enhancer regulation in heroin addiction by studying the effect of bidirectional manipulations of BRD4 in NAc and DS on gene expression and behavioral endpoints in rat models, including screening several BRD4/BET inhibitors for anti-addiction actions. These studies offer a translational opportunity to advance novel therapies for drug addiction.