Deciphering the single-nucleus genomic regulatory structure of opioid use disorder in the human brain

项目摘要 阿片类药物使用障碍(OUD)在美国已达到流行水平，与 住院和药物过量死亡，后者在2020年大幅上升，最高可达29.4%。 虽然遗传风险因素已经在最近的大规模全基因组关联研究中被识别出来。 当然，这些只解释了部分差异，并且通常映射到非编码区。表观遗传修饰 与基因和环境下的阿片使用障碍(OUD)的病因学有牵连 相互影响。我们和其他人发现，DNA甲基化(5mC)的变化是研究最多的 表观遗传机制，与人类外周和死后脑中的OUD有关。然而， 这项工作的大部分都是在块状组织中完成的，这掩盖了细胞多样性在 人类细胞。此外，还需要研究来评估额外的和新的表观遗传调节层，以获得 更好地理解它对基因调控的贡献以及它解释GWAs功能的能力 OUD背景下的遗传变异。在这里，我提供了一个新的框架来解决这些差距和挑战：1) 同时对DNA甲基化、DNA羟甲基化和3D基因组结构进行分析 单个人核，2)识别细胞类型和脑区域内的OUD依赖的调节信号，3) 评估不同表观基因组调节层之间的串扰，以及4)构建基因程序以 精细地映射oud Gwas变体和多基因信号以发挥作用。这种综合性的单细胞多细胞组学 OUD标测将检查背外侧前额叶皮质(DLPFC)、杏仁核(BLA)和核团 伏隔(NAcc)，成瘾回路的一部分，从人类死后大脑样本中收集 UTHealth大脑收集数据集，并使用退伍军人脑库(Npbb)作为验证队列。这项工作是 极具创新性，将开辟新的研究路线，通过提供新颖的 对其在人脑中的基因调控结构的机械论见解。这项拟议的研究将确定和 帮助告知分子靶点，用于预防和治疗患有 奥德。

Project Summary Opioid use disorder (OUD) has reached epidemic levels in the Unites States, associated with increased rates of hospitalization and drug overdose death, the latter showing a significant steep rise of up to 29.4% in 2020. While genetic risk factors have been identified in recent large-scale genome-wide association studies (GWAS) of OUD, these explain only part of the variance and often map to noncoding regions. Epigenetic modifications have been implicated in the etiology of opioid use disorders (OUD) underlying the gene and environment interplay. We and others have found that alterations of DNA methylation (5mC), one of the most studied epigenetic mechanisms, is associated with OUD in both human peripheral and postmortem brain. However, most of this work has been done in bulk tissues, which obscures the functional role of the cellular diversity in human cells. Further, research is needed to assess additional and novel epigenetic regulatory layers to gain a better understanding of its contribution to gene regulation and its ability to interpret the functionality of GWAS genetic variants in the context of OUD. Here, I offer a novel framework to tackle these gaps and challenges: 1) conduct a simultaneous profiling of DNA methylation, DNA hydroxymethylation, and 3D genome structure in single human nuclei, 2) identify OUD-dependent regulatory signatures within cell types and brain regions, 3) evaluate the crosstalk between the different epigenomic regulatory layers, and 4) construct gene programs to finely map OUD GWAS variants and polygenic signals to function. This comprehensive single-cell multiomics mapping of OUD will examine the dorsolateral prefrontal cortex (DLPFC), amygdala (BLA), and nucleus accumbens (NAcc), part of the addiction circuitry, of human postmortem brain samples collected from the UTHealth Brain Collection datasets and using the VA Brain Bank (NPBB) as a validation cohort. This work is highly innovative and will open new lines of research the genetics and epigenetics of OUD by providing novel mechanistic insights on its gene regulatory structure in the human brain. This proposed study will identify and help inform molecular targets to be used as prevention and treatment efforts for individuals suffering from OUD.