Molecular rhythm alterations in human post-mortem brain associated with opioid use disorder

项目摘要 在美国，阿片类药物的使用和依赖流行率急剧上升。大多数阿片类药物患者 使用障碍（OUD）在治疗后数月内复发。最近的人类神经影像学和死后 OUD的脑研究揭示了皮质和纹状体脑回路功能障碍的程度，特别是在 背外侧前额叶皮层（DLPFC）和背外侧前额叶核（NAc）区域，与阿片样物质密切相关 使用和依赖风险。前额叶皮层为前额叶皮层提供自上而下的抑制性认知和情绪控制， 它调节目标导向和奖励行为。OUD的复发脆弱性与以下因素密切相关： 睡眠和昼夜节律中断的严重性和持续性，提高了治疗性 在戒断期间减轻这些干扰的干预措施可能有效减少阿片类药物的渴望 和复发然而，我们对生物学机制的理解， 昼夜节律和OUD之间的联系是有限的，特别是在OUD患者大脑的分子水平上。我们 和其他人已经开发了新颖的，创新的方法，使用死亡时间（TOD）来测量分子水平。 节奏在人类死后的大脑，以调查物质使用之间的机械联系， 分子脑节律。使用TOD方法，我们最近发现， 与正常衰老和精神疾病相关的前额皮质。值得注意的是，我们还发现， 疾病特异性分子通路内基因的节律性，为大脑生物学提供了新的见解 衰老和精神病理学。人类受试者大规模基因表达的初步TOD分析 与OUD显示富集与PFC和NAc的昼夜节律相关的途径。在我们的提议中， 我们将直接研究分子节律破坏与阿片类药物使用和复发之间的关系， 使用来自OUD受试者的人类死后大脑和电路特异性靶向的小鼠模型 和阿片类药物自我给药。具体来说，我们将研究死后DLPFC的分子节律， 使用RNA测序从OUD受试者的大队列中获得NAc（Aim 1A）。我们还将研究 特定临床特征（例如，毒理学报告和药物过量、共病精神疾病、病史 使用、多种药物使用、病程）对OUD分子节律的影响（目的1B）。然后我们将直接测试 特定脑区（PFC和NAc; Aim 2A）和回路中分子节律中断的功能相关性 (PFC投射到NAc;目的2B）。我们的研究将确定 OUD受试者大脑中的分子节律异常，开始确定其机制 将昼夜节律和成瘾联系起来，这将为疾病相关途径提供重要的见解， 也是潜在的治疗策略。

PROJECT SUMMARY Opioid use and dependence prevalence have skyrocketed in the United States. A majority of patients with opioid use disorder (OUD) relapse within months despite treatment. Recent human neuroimaging and postmortem brain studies in OUD reveal the degree of dysfunction within cortical and striatal brain circuits, particularly within dorsolateral prefrontal cortical (DLPFC) and nucleus accumbens (NAc) regions, strongly relates to the opioid use and dependence risk. The PFC provides top-down inhibitory cognitive and emotional control to the NAc, which mediates goal-directed and reward behaviors. Relapse vulnerability in OUD is strongly associated with the severity and persistency of disruptions to sleep and circadian rhythms, raising the possibility that therapeutic interventions which mitigate these disruptions during abstinence may be effective for reducing opioid craving and relapse. However, our understanding of the biological mechanisms underlying the relationships between circadian rhythms and OUD is limited, especially at the molecular level in the brains of people with OUD. We and others have developed novel, innovative approaches using time of death (TOD) to measure molecular rhythms in the human postmortem brain to investigate the mechanistic links between substance use and molecular brain rhythms. Using TOD approaches, we recently found a marked loss of molecular rhythms in the prefrontal cortex associated with normal aging and psychiatric disorders. Notably, we also discovered a gain of rhythmicity in genes within disease-specific molecular pathways, providing novel insights into the biology of brain aging and psychiatric pathology. Preliminary TOD analyses on large-scale gene expression in human subjects with OUD revealed enrichment for pathways related to circadian rhythms in the PFC and NAc. In our proposal, we will directly investigate the relationship between molecular rhythm disruption and opioid use and relapse using both human postmortem brains from subjects with OUD and mouse models of circuit-specific targeting and opioid self-administration. Specifically, we will investigate molecular rhythms in postmortem DLPFC and NAc using RNA-sequencing from a large cohort of subjects with OUD (Aim 1A). We will also examine the impact of specific clinical features (e.g., toxicology reports and overdoses, comorbid psychiatric disorders, history of use, polysubstance use, illness duration) on molecular rhythms in OUD (Aim 1B). We will then directly test the functional relevance of molecular rhythm disruptions in specific brain regions (PFC and NAc; Aim 2A) and circuits (PFC projections to NAc; Aim 2B) during opioid self-administration behavior in mice. Our studies will identify molecular rhythm abnormalities in the brains of subjects with OUD and begin to determine the mechanisms linking circadian rhythms and addiction, which will provide important insight into disease-related pathways and also potential treatment strategies.