Role of Cell Type-Specific Molecular Rhythm Disruption in Alcohol Use Disorder

细胞类型特异性分子节律破坏在酒精使用障碍中的作用

• 批准号: 10725280
• 负责人: Kyle Ketchesin
• 金额: $ 44.05万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-14 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10725280
• 关键词: ARNTL gene; Ablation; Alcohol consumption; Astrocytes; Autopsy; Behavior; Behavioral; Brain; Cells; Cessation of life; Chronic; Circadian Dysregulation; Circadian Rhythms; Corpus striatum structure; Data; Disease; Dopamine Receptor; Enterobacteria phage P1 Cre recombinase; Feedback; Future; Gene Expression; Genes; Genetic Polymorphism; Genetic Transcription; Health; Heavy Drinking; Hour; Human; Individual; Injury; Link; LoxP-flanked allele; Maps; Measures; Mediating; Mental disorders; Molecular; Mus; Neurons; Nucleus Accumbens; Patients; Pattern; Periodicity; Physiological; Play; Population; Public Health; Regulation; Rewards; Risk; Rodent; Role; Sleep; Sleep Wake Cycle; Testing; Time; Tissues; Transcription Alteration; Viral; World Health Organization; addiction; alcohol exposure; alcohol use disorder; burden of illness; cell type; circadian; circadian pacemaker; circadian regulation; comparison control; drinking; drinking behavior; human RNA sequencing; innovation; insight; molecular clock; mouse model; new therapeutic target; novel therapeutics; promoter; single nucleus RNA-sequencing; substance use; time use; tool; transcriptome; treatment strategy

PROJECT SUMMARY/ABSTRACT Alcohol use is a causal factor in over 200 health conditions and is estimated to account for 5.1% of global burden of disease and injury. Despite the enormous public health impact, we still lack a basic understanding of the mechanisms that contribute to alcohol use disorder (AUD). A prominent feature of AUD is circadian rhythm disturbances, including altered physiological rhythms and sleep/wake cycles. The relationship is bidirectional, as rhythm disruptions can exacerbate alcohol consumption and polymorphisms in canonical circadian genes associate with increased alcohol use. However, very little is known about the mechanisms underlying these relationships, especially at the molecular level in the brains of individuals with AUD. Molecular rhythms are generated by transcriptional-translational feedback loops to control circadian-dependent (near 24-hour) gene expression. Notably, AUD is associated with disrupted molecular rhythms of canonical circadian genes in the periphery and rodent studies have shown that disruptions to molecular rhythms in the striatum, particularly the nucleus accumbens (NAc), are associated with altered reward regulation and increased risk for substance use. Measures of molecular rhythms in human postmortem brain, however, have historically been challenging since each brain represents a single circadian timepoint. An innovative analysis uses “time of death” (TOD) to fit subjects on a 24-hr ‘clock’; by combining expression data from all subjects, it is possible to reconstruct molecular rhythm patterns in the human postmortem brain. To investigate AUD-associated molecular rhythm alterations, we performed a preliminary study examining large-scale transcriptional changes in bulk NAc tissue from subjects with AUD compared to controls. Notably, core circadian genes (e.g., CRY1, PER2), which display rhythmic expression in control subjects, were arhythmic in AUD, suggesting a molecular link to behavioral rhythm alterations observed in AUD patients. However, the NAc is composed of a variety of transcriptionally distinct cell types with differential roles in alcohol drinking behavior, and the role of molecular rhythms in these cell types has not been previously investigated. The central hypothesis of this R21 proposal is that molecular rhythms are disrupted in a cell-type specific manner in the NAc of AUD subjects, and that these disruptions contribute to excessive drinking. To test this hypothesis, we will use single nucleus RNA-sequencing (snRNA-seq) of human postmortem NAc tissue to determine how molecular rhythms are altered in heterogenous cell populations in AUD (Aim 1). We will then manipulate rhythms in specific NAc cell types to determine a causal role for rhythm disruption in alcohol drinking (Aim 2). Together, these studies will be the first to examine cell type-specific rhythms in human striatum, how they are altered in AUD, and their contribution to excessive alcohol consumption. These studies are central to understanding the mechanisms underlying circadian disruptions in AUD and may result in the discovery of novel therapeutic targets and/or time-dependent strategies for future treatments.

项目摘要/摘要 饮酒是200多个健康状况中的因果因素，估计占全球负担的5.1％ 疾病和伤害。尽管公共卫生产生了巨大的影响，但我们仍然缺乏对 导致饮酒障碍的机制（AUD）。 AUD的一个突出特征是昼夜节律 干扰，包括改变生理节奏和睡眠/唤醒周期。关系是双向的，如 节奏干扰会加剧饮酒和多态性的昼夜节律基因的多态性 与饮酒增加相关。但是，关于这些机制的知识知之甚少 关系，尤其是在具有AUD的个体的大脑的分子水平上。分子节奏为 由转录 - 翻译反馈循环产生以控制昼夜节律依赖性（接近24小时）基因 表达。值得注意的是，AUD与典型昼夜节律基因的分子节律有关 周围和啮齿动物的研究表明，纹状体中的分子节律破坏，尤其是 伏隔核（NAC）与奖励调节的改变和使用物质的风险增加有关。 但是 每个大脑代表一个昼夜节律的时间点。创新的分析使用“死亡时间”（TOD）适合 24小时“时钟”的受试者；通过结合所有受试者的表达数据，可以重建分子 人类验尸大脑中的节奏模式。为了研究听觉相关的分子节律改变， 我们进行了一项初步研究，研究了受试者的大规模NAC组织的大规模转录变化 与对照相比，AUD。值得注意的是，核心昼夜节律基因（例如Cry1，per2），显示有节奏的 在对照受试者中的表达在AUD中是心律不振的，这表明与行为节奏的分子联系 在AUD患者中观察到的改变。但是，NAC由各种转录不同的细胞组成 在饮酒行为中具有不同作用的类型，分子节奏在这些细胞类型中的作用具有 以前没有研究过。该R21提案的中心假设是分子节奏是 在AUD受试者的NAC中以细胞类型的特定方式破坏，这些中断有助于 酗酒。为了检验这一假设，我们将使用人类的单核RNA测序（SnRNA-Seq） 验尸NAC组织，以确定AUD异质细胞种群中分子节律的改变 （目标1）。然后，我们将在特定的NAC细胞类型中操纵节奏以确定节奏的因果作用 饮酒的破坏（AIM 2）。总之，这些研究将是第一个检查细胞类型特异性的研究 人类纹状体的节奏，AUD的改变以及它们对过量饮酒的贡献。 这些研究对于理解AUD昼夜节律中断的机制至关重要，可能 导致发现新的治疗靶标和/或时间依赖于未来治疗的策略。