Multiomic profiling of cell types mediating opioid use disorder in rats

介导大鼠阿片类药物使用障碍的细胞类型的多组学分析

• 批准号: 10681460
• 负责人: Giordano De Guglielmo
• 金额: $ 55.8万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-15 至 2027-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10681460
• 关键词: Acute; Addictive Behavior; Affect; Amygdaloid structure; Analgesics; Animal Model; Atlases; Behavioral; Behavioral Model; Biological; Biological Assay; Brain; Brain region; CRISPR/Cas technology; Candidate Disease Gene; Cell Nucleus; Cells; Chromatin; Chronic; Clustered Regularly Interspaced Short Palindromic Repeats; Collaborations; Computer Analysis; Cue-induced relapse; Data; Data Set; Development; Disease; Disease Progression; Drug Kinetics; Drug abuse; Drug usage; Epigenetic Process; Exposure to; Female; Fentanyl; Funding; Future; Gene Expression; Gene Expression Profiling; Genes; Genetic Transcription; Genome; Genomics; Goals; Grant; Habenula; Heroin; Human; Hypothalamic structure; In Situ; In Situ Hybridization; Individual; Intravenous; Investigation; Knowledge; Lateral; Link; Maps; Measurement; Measures; Mediating; Messenger RNA; Methodology; Modeling; Molecular; National Institute of Drug Abuse; Neuroanatomy; Nucleus Accumbens; Opiate Addiction; Opioid; Overdose; Oxycodone; Pain Disorder; Prefrontal Cortex; Prevention; Process; Protocols documentation; Public Health; RNA; Rattus; Regulatory Element; Research; Resolution; Risk Factors; Rodent Model; Role; Saline; Sampling; Scientist; Self Administration; Substance abuse problem; System; Technology; Time; Tissues; Transcriptional Regulation; United States; Validation; Viral; Vulnerable Populations; Withdrawal; XCL1 gene; addiction; behavioral phenotyping; brain cell; brain tissue; cell type; clinically relevant; drug relapse; epigenome; epigenomics; gene regulatory network; genomic data; illicit opioid; in vivo; innovation; knock-down; long term recovery; male; multimodality; multiple omics; neuroadaptation; opioid epidemic; opioid exposure; opioid misuse; opioid mortality; opioid overdose; opioid use; opioid use disorder; prescription opioid misuse; prescription pain reliever; prolonged abstinence; response; synthetic opioid; transcriptome; transcriptomics

Project Summary The misuse and abuse of prescription pain relievers, such as oxycodone, contributed to the unprecedented opioid epidemic in the United States. The opioid crisis has devastating consequences on public health including a surge in opioid misuse and related overdoses. Research is urgently needed to develop better treatments for opiate addiction. Despite substantial knowledge of the pharmacokinetic and behavioral effects of oxycodone in various animal models, only a small number of candidate genes and neuroanatomical systems affected by opioids have been studied. Recent technological advances in the field of single cell genomics are promising avenues for the unbiased discovery and characterization of brain cell types that respond to opioids. In response to this RFA, we leverage an innovative multi-omics methodology (Single Cell Multiome ATAC + Gene Expression) to map the transcriptome and epigenome from the same cell across thousands of cells in brain regions relevant to the effects of opioid exposure. To this aim we will use a rat model of extended access to oxycodone intravenous self-administration that recapitulates several neuroadaptations also observed in humans with opioid use disorders (OUD). This approach provides an exceptional opportunity to systematically explore the cellular diversity of the opioid system and, at the same time, the causative mechanisms that regulate cellular states based on the associations between epigenetic changes and the expression of target genes in individual cells. We will integrate this innovative multi-omics methodology with rigorous computational approaches to explore the cellular organization the opioid system in multiple brain regions and different stages of OUD progression (initial exposure, escalation of use, acute withdrawal, prolonged abstinence, and cue- induced relapse). We have provided strong preliminary that support the feasibility of our proposed plan for the following aims. In Aim 1, we will collect brain tissues at different stages of the extended access to oxycodone intravenous self-administration (ivsa) protocol and we will generate single cell genomics data from both male and female rats that are exposed to either saline or oxycodone. In Aim 2, we will integrate these transcriptomic and epigenomic datasets to identify changes in cellular states, genes and upstream regulators that are associated with different stages of oxycodone use. This approach will facilitate the identification of linkages between cis- regulatory elements and target genes. In Aim 3, we will validate key cell type-specific findings by RNA-FISH and identify the top 3 target genes for functional validation. To this aim, we will use a viral-mediated CRISPR- Cas9 system to modulate addictive behaviors in rat models of oxycodone self-administration. The results of this study will enable future studies that may identify new targets for treatment and prevention of OUD.

项目摘要 滥用和滥用处方止痛药（例如羟考酮）为前所未有的 美国的阿片类流行病。阿片类药物危机对公共卫生有毁灭性后果 阿片类药物滥用和相关过量的激增。迫切需要进行研究以开发更好的治疗方法 鸦片成瘾。 尽管有大量了解羟考酮在各种的药代动力学和行为影响 动物模型，只有少数受阿片类药物影响的候选基因和神经解剖学系统具有 被研究。单细胞基因组学领域的最新技术进步是有希望的途径 对阿片类药物反应的脑细胞类型的无偏见和表征。 为了响应这种RFA，我们利用创新的多摩学方法（单细胞多组件 ATAC +基因表达）以绘制来自同一细胞的转录组和表观基因组的数千个 大脑区域的细胞与阿片类药物暴露的作用有关。为此，我们将使用扩展的大鼠模型 访问羟考酮静脉自我给药，这些自我给药概括了几种神经适应 患有阿片类药物使用障碍的人（OUD）。这种方法为系统地提供了出色的机会 探索阿片类药物系统的细胞多样性，同时探索调节的致病机制 细胞状态基于表观遗传变化与靶基因表达之间的关联 单个细胞。我们将将这一创新的多词方法与严格的计算集成 探索细胞组织的方法在多个大脑区域和不同阶段 OUD进展（初始暴露，使用升级，急性戒断，戒酒延长和提示 - 诱发复发）。 我们提供了强大的初步，以支持我们提议的计划的可行性 目标。在AIM 1中，我们将在扩展进入羟考酮静脉内的不同阶段收集脑组织 自我管理（IVSA）方案，我们将生成男性和女性的单细胞基因组数据 暴露于盐水或羟考酮的大鼠。在AIM 2中，我们将整合这些转录组和 表观基因组学数据集，以识别与细胞状态，基因和上游调节剂的变化相关的变化 羟考酮的不同阶段使用。这种方法将促进识别顺式之间的联系 调节元素和靶基因。在AIM 3中，我们将通过RNA-Fish验证钥匙细胞类型特异性发现 并确定用于功能验证的前3个目标基因。为此，我们将使用病毒介导的CRISPR- CAS9系统以调节羟考酮自我管理大鼠模型中的上瘾行为。结果的结果 研究将使未来的研究能够确定用于治疗和预防OUD的新目标。