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

基本信息

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
项目类别：
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

项目摘要

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.

项目摘要美国面临着由滥用和滥用造成的前所未有的阿片类药物流行处方止痛药和非法阿片类药物。这个问题对公共卫生有毁灭性的后果，包括过量药物相关的死亡，新生儿戒断综合征和扩散的显着增加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）具有强遗传成分。但是，与阿片类药物成瘾有关的基因网络仍然很差理解。该提案的主要目的是研究转录调节机制是羟考酮滥用障碍的不同阶段的发展。我们将利用定量表观基因组方法将提供一个综合的调节元素图，转录沿OUD轨迹在特定阶段失调的因素和下游靶基因。我们的主要创新是使用限制的小（CS）RNA-Seq，我们开发了一种量化新近量化的方法直接从总RNA直接具有高灵敏度和高空间分辨率的启动转录本。这种方法实现激活基因的转录起始位点（TSS）的公正注释和转录单核苷酸分辨率下的调节元件。与其他表观基因组分析相比，CSRNA-SEQ为对转录的变化高度敏感，它可以捕获稳定基因和不稳定的转录本，例如增强子RNA。为了研究OUD不同阶段的监管变化，我们将在扩展接入条件下使用羟考酮自我给药的大鼠模型。该模型概括了人类成瘾行为的几个方面，包括宽容，依赖和动机。因此，它增强了我们结果的翻译相关性。该提案将使用两个表现大的近交菌株他们在禁欲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暴露于羟考酮。结合使用转录启动分析其他敏感的分析技术，我们将研究基础的转录调节网络 OUD的不同阶段，包括初始暴露，使用升级，急性和持续戒酒。我们提出的研究一起将通过定义监管网络对现场产生广泛的影响与沿着大鼠OUD轨迹不同阶段的脆弱性相关的表型的基础，它可能导致新的治疗靶标治疗OUD。