Three Dimensional Chromosome Architecture in Drug Addiction

项目摘要/摘要 神经功能需要准确控制基因转录以响应环境刺激。 基因表达异常被认为是导致药物成瘾的原因。因此，研究基因的调控 药物成瘾的表达可能会为这种疾病提供机械性的洞察，但这种疾病提供的信息仍然有限 可供选择的治疗方法，是巨大的社会和经济负担。据估计，有多达 哺乳动物基因组中的一百万个调控元件，它们通常位于很远的地方 它们的目标基因。尽管下一代测序的最新进展为大规模的 对调控DNA元件的识别，即它们调控哪些基因，在很大程度上仍是未知的。最近， 线状DNA的有序紧凑和组织进入细胞核已被认为具有 促进基因启动子与远端相互作用对基因转录的主要影响 监管要素。然而，这种三维染色质结构是如何在大脑中组织的 它是如何改变的，特别是在吸毒成瘾方面，仍然是个谜。在这里，我们建议研究长期的- 远端DNA元件与所有带注释的小鼠基因启动子之间的范围环相互作用 特定的神经元亚型不同地参与成瘾。为了实现这一点，我们将应用切割 边缘启动子捕获Hi-C技术在D1-和D2-培养基中对整个启动子互作体谱的研究 伏隔核中的棘神经元(MSN)是大脑奖励的关键结构。我们还将研究 可卡因和海洛因成瘾小鼠神经元亚型特异性高阶基因组组织。我们 预期阐明神经元亚型特异性和药物特异性的三维染色体 架构发生了变化。这些变化中的许多可能涉及以前发现的遗传变异位点 在啮齿动物和人类之间保存。对这些监管区域的操纵不仅可能改变 调节其靶基因的表达，但也可能改变与之相关的成瘾行为。vt.在.的基础上 完成后，我们的研究不仅将无可争辩地推动我们对药物成瘾的理解 尚未实现的基因组架构组织维度，它也将开辟一条新的途径 对疾病的操纵，这对未来的治疗应用具有潜在的实用价值。

PROJECT SUMMARY / ABSTRACT Neural function requires accurate control of gene transcription in response to environmental stimuli. Aberrant gene expression is believed to drive drug addiction. Therefore, studying the regulation of gene expression in drug addiction may provide mechanistic insights to this disease, which still offers limited options for treatment and represents a vast social and economic burden. It is estimated that there are up to one million regulatory elements in the mammalian genome, which are often located great distances from their target genes. Although recent developments in next generation sequencing have provided large-scale identification of regulatory DNA elements, which genes they regulate remains largely unknown. Recently, the ordered compaction and organization of linear DNA into the nucleus has been recognized as having a major influence on gene transcription by facilitating interactions between gene promoters and distal regulatory elements. However, how this three-dimensional chromatin architecture is organized in the brain and how it is changed, particularly in drug addiction, is still obscure. Here we propose to study the long- range looping interactions between distal DNA elements and all annotated mouse gene promoters within the specific neuronal subtypes differentially engaged in addiction. To achieve this, we will apply the cutting edge promoter capture Hi-C technology to profile the entire promoter interactome in D1- and D2- medium spiny neurons (MSNs) in nucleus accumbens, the key brain reward structure. We will also examine the neuron subtype-specific higher order genome organization in both cocaine and heroin addicted mice. We anticipate to elucidate neuron subtype specific and drug specific three-dimensional chromosome architecture changes. Many of these changes may involve previously identified genetic variation sites conserved between rodents and human. Manipulation of these regulatory regions may not only alter regulation of their target gene's expression, but may also change the associated addiction behaviors. Upon completion, our study will not only indisputably advance our understanding of drug addiction to the unachieved dimension of genome architecture organization, it will also open a new avenue to a plausible manipulation of the disease, which has potential utility for future therapeutic applications.