Dynamic epigenomic landscape of opioid abuse following early-life adversity

项目总结 在过去的十年里，美国出现了毁灭性的阿片成瘾流行，致命的阿片类药物过量 发病率在10年内翻了一番。遗传因素显然是成瘾易感性的原因，但它们本身不能 这是造成这种增长的原因。因此，环境因素也必须发挥作用，其持久影响将是 在生命早期的敏感/关键时期达到最高水平。事实上，早期生活逆境(ELA)与阿片类药物有关 上瘾的脆弱性，在实验模型中，它促进了这种脆弱性。尽管负有责任的 机制尚未阐明，ELA启动的变化的持久性质强烈表明 表观遗传机制可能是这些稳定变化的基础。表观基因组是一个信号整合平台 叠加在基因组上，基因组被ELA和滥用药物修饰，还编码 性别差异的各个方面。已知的表观遗传机制建立了细胞功能的稳定变化(通过 协调基因调控)导致持续改变的行为，包括吸毒和寻求毒品 行为。因此，了解与成瘾倾向相关的表观遗传和转录变化 在ELA之后可能会回答关于阿片类药物使用障碍的原因和机制的几个关键问题 (有声)。例如，ELA是如何表观遗传编码的，从而增加了阿片类药物滥用和复发的风险- 相关行为？海洛因对ELA患者的表观基因组和转录组是否有不同的影响 历史，宣扬这种脆弱性？这些问题将在这里使用强大的ELA和ELA大鼠模型来解决 海洛因的自我给药和复发。利用我们的发现，ELA增加了海洛因的易感性和 与人类数据20-23一致，我们将检验驱动假说，即 ELA和最初的海洛因使用调节表观基因组过程，这些过程反过来协调基因调节， 最终影响细胞信号和OUD样行为。表观基因组学和转录组学变化 将采用整体和单细胞多组学方法确定ELA和海洛因诱导的 OUD轨迹中不同点的大脑区域。此外，还将采集脑脊液和血液样本。 细胞外小泡(EV)miRNA图谱能够潜在开发OUD风险的预测标记物 和/或进展。使用最先进的测序和分析方法，我们将揭示基因调控 与ELA有关的网络、海洛因的初次使用以及女性和男性的海洛因滥用/复发。这种方法 这将使我们能够确定预防和干预弱势群体的新目标。

PROJECT SUMMARY The last decade has seen a devastating epidemic of opioid addiction in the U.S with the fatal opioid overdose incidence doubling in 10 years. Genetic factors clearly contribute to addiction vulnerability, but they alone cannot account for this rise. Therefore, environmental factors must also play role, and their enduring effects would be maximal during sensitive/critical periods early in life. Indeed, early life adversity (ELA) is associated with opioid addiction vulnerability, and in experimental models, it promotes such vulnerability. Although the responsible mechanisms have yet to be elucidated, the enduring nature of changes set in motion by ELA strongly suggest that epigenetic mechanisms may underlie these stable changes. The epigenome is a signal integration platform superimposed on the genome, which is modified by both ELA and drugs of abuse, and which also encodes aspects of sex differences. Epigenetic mechanisms are known to establish stable changes in cell function (via coordinating gene regulation) that give rise to persistently altered behaviors, including drug use and drug seeking behaviors. Thus, understanding epigenetic and transcriptional changes associated with addiction propensity following ELA may answer several key questions about the causes and mechanisms of opioid use disorder (OUD). For example, how is ELA epigenetically encoded, thereby increasing risk for opioid abuse and relapse- associated behaviors? Does heroin differentially affect the epigenome and transcriptome in those with ELA history, promoting this vulnerability? These questions will be addressed here using robust rat models of ELA and of heroin self-administration and relapse. Capitalizing on our findings that ELA augments heroin vulnerability and relapse in a sex-divergent manner4, consistent with human data20-23, we will test the driving hypothesis that both ELA and initial heroin use regulate epigenomic processes, and these, in turn, coordinate gene regulation that ultimately affects cellular signaling, and OUD-like behaviors. The epigenomic and transcriptomic changes induced by ELA and heroin will be determined by employing both bulk- and single-cell multi-omics in selected brain regions at different points in the OUD trajectory. They will be complemented by sampling of CSF and blood extracellular vesicle (EV) miRNA profiles to enable potential development of predictive markers of OUD risk and/or progression. Using state of the art sequencing and analysis methods, we will uncover gene regulatory networks associated with ELA, initial heroin use, and heroin abuse/relapse in females and males. This approach will allow us to identify novel targets for OUD prevention and intervention in vulnerable individuals.