Decoding the grammar of transcriptional enhancers regulating different stages of opioid use disorder

解码调节阿片类药物使用障碍不同阶段的转录增强子的语法

• 批准号: 10269005
• 负责人: Christopher W Benner
• 金额: $ 70.51万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-30 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10269005
• 关键词: ATAC-seq; Abstinence; Acute; Address; Affect; American; Area; Atlases; Behavior; Behavioral; Behavioral Model; Binding; Biochemical Pathway; Brain; Brain region; Cell Nucleus; Chromatin; Communicable Diseases; Complex; Computer Analysis; Computing Methodologies; Data; Dependence; Development; Disease; Drug Kinetics; Drug Prescriptions; Drug abuse; Drug usage; Enhancers; Epigenetic Process; Exhibits; Exposure to; Female; Gene Expression; Gene Expression Regulation; Gene Targeting; Genes; Genetic; Genetic Enhancer Element; Genetic Transcription; Genome; Genomic Segment; Genomics; Goals; HIV; Hepatitis; Heritability; Heroin; Hi-C; Human; Human Genetics; Illicit Drugs; Inbred Strain; Inbred Strains Rats; Intake; Intoxication; Lead; Maps; Methods; Modeling; Molecular; Motivation; Neonatal Abstinence Syndrome; Nucleotides; Opiate Addiction; Opioid; Overdose; Oxycodone; Pain; Phenotype; Protocols documentation; Public Health; RNA; Rat Strains; Rattus; Regulation; Regulator Genes; Regulatory Element; Resolution; Risk; Rodent Model; Role; Sampling; Scientist; Self Administration; Small Nuclear RNA; Small RNA; Techniques; Technology; Time; Tissues; Transcript; Transcription Initiation; Transcription Initiation Site; Transcriptional Regulation; Translating; United States; Untranslated RNA; Variant; Ventilatory Depression; addiction; base; behavioral phenotyping; behavioral study; brain tissue; cell type; cohort; deep sequencing; epigenomics; genetic approach; genetic variant; genome wide association study; genome-wide; genomic data; human study; innovation; male; new therapeutic target; novel; opioid epidemic; opioid use disorder; prescription opioid; prescription pain reliever; response; screening; sex; therapeutic development; trait; transcription factor; transcriptome sequencing

Project Summary The United States is facing an unprecedented opioid epidemic caused by the misuse and abuse of both prescription pain relievers and illegal opioids. This issue has devastating consequences for public health, including a significant increase in overdoses related fatalities, in neonatal withdrawal syndrome and spread of infectious diseases such as HIV and hepatitis. Numerous studies indicate that opioid use disorder (OUD) has a strong genetic component. However, the genes networks implicated in opioid addiction remain poorly understood. The primary goal of this proposal is to study the transcriptional regulatory mechanisms that underlie the development of distinct stages of oxycodone abuse disorder. We will leverage the power of quantitative epigenomic methods that will provide a comprehensive map of regulatory elements, transcription factors and downstream target genes that are dysregulated in specific stages along the OUD trajectories. Our major innovation is the use of capped small (cs)RNA-seq, a method that we developed to quantify newly initiated transcripts with high sensitivity and high spatial resolution directly from total RNA. This approach enables the unbiased annotation of Transcriptional Start Sites (TSS) of both activated genes and transcribed regulatory elements at single nucleotide resolution. Compared with other epigenomic profiling, csRNA-seq is highly sensitive to changes in transcription, and it can capture the dynamic regulation of both stable genes and unstable transcripts, such as enhancer RNA. To study regulatory changes in distinct stages of OUD, we will use a rat model of oxycodone self-administration under extended access conditions. This model recapitulates several aspects of the human addiction-like behaviors, including tolerance, dependence, and motivation. Thus, it enhances the translational relevance of our results. This proposal will use two inbred strains that exhibit large differences in their motivation to seek oxycodone during abstinence while having similar pharmacokinetic for oxycodone and similar exposure to oxycodone. Using transcriptional initiation profiling, in combination with other sensitive profiling techniques, we will investigate the transcriptional regulatory networks underlying different stages of the OUD, including initial exposure, escalation of use, acute and sustained abstinence. Together, our proposed studies will have a broad impact in the field by defining regulatory networks that underlie phenotypes associated with vulnerability to distinct stages along the OUD trajectory in rats, and it may lead to novel therapeutic targets to treat OUD.

项目摘要 美国正面临一场史无前例的阿片类药物流行，这是由两者的误用和滥用造成的 处方止痛药和非法阿片类药物。这一问题对公共卫生造成了毁灭性的后果， 包括与过量服药有关的死亡人数、新生儿戒断综合症和 艾滋病毒和肝炎等传染病。大量研究表明，阿片类药物使用障碍(OUD)具有 很强的遗传成分。然而，与阿片成瘾有关的基因网络仍然很差。 明白了。这项提案的主要目标是研究转录调控机制， 是羟考酮滥用障碍不同阶段发展的基础。我们将利用 定量的表观基因组学方法，将提供调控元件、转录的全面图谱 因子和下游靶基因在特定阶段沿OUD轨迹不受调控。我们的 主要的创新是使用封顶小(Cs)rna-seq，这是我们开发的一种新的量化方法。 直接从总RNA中启动具有高灵敏度和高空间分辨率的转录本。这种方法 允许对激活的基因和转录的基因的转录起始位点(TS)进行无偏见的注释 单核苷酸分辨率的调控元件。与其他表观基因组图谱相比，csRNA-seq 对转录变化高度敏感，并能捕捉稳定基因和 不稳定的转录本，如增强子RNA。为了研究OUD不同阶段的监管变化，我们将 在延长访问条件下使用羟考酮自我给药的大鼠模型。这个模型概括了 人类成瘾行为的几个方面，包括容忍、依赖和动机。因此， 它增强了我们结果的翻译相关性。这项提案将使用两个近交系菌株，它们表现出较大的 戒毒期间寻求羟考酮的动机差异，同时具有相似的药代动力学 羟考酮和类似的羟考酮暴露。使用转录启动特征分析，结合 其他敏感的分析技术，我们将研究潜在的转录调控网络 OUD的不同阶段，包括最初的暴露、使用的升级、急性和持续的戒断。 总之，我们拟议的研究将通过定义以下监管网络在该领域产生广泛影响 与大鼠对OUD轨迹不同阶段的易感性相关的表型，可能 导致新的治疗靶点来治疗OUD。