Role for Circular RNAs in Compulsive Cocaine Intake

环状 RNA 在强迫性可卡因摄入中的作用

• 批准号: 10306369
• 负责人: Paul J. Kenny
• 金额: $ 26.7万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-02-15 至 2023-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10306369
• 关键词: Abstinence; Autopsy; Aversive Stimulus; Awareness; Back; Behavior; Behavioral; Biological Assay; Biological Process; Brain; Brain region; CRISPR/Cas technology; Cells; Chromatin; Clustered Regularly Interspaced Short Palindromic Repeats; Cocaine; Cocaine Dependence; Cocaine use disorder; Collaborations; Complement; Consumption; Corpus striatum structure; Data; Development; Disease; Dopamine D1 Receptor; Dorsal; Dose; Drug Addiction; Etiology; Functional disorder; Future; Genes; Genetic Transcription; Goals; Habits; Human; In Situ Hybridization; Intake; Investigation; Laboratories; Laboratory Animals; Mammalian Cell; Mediating; Methods; MicroRNAs; Molecular; Motivation; Mus; Nature; Neurobiology; Neuroglia; Neuronal Plasticity; Neurons; Nucleus Accumbens; Pattern; Persons; Pharmaceutical Preparations; Play; Population; Program Research Project Grants; Property; Proteins; Public Health; RNA; RNA Interference; RNA Splicing; Regulation; Relapse; Role; Self Administration; Site; Slice; Substance Use Disorder; Synapses; Synaptic Transmission; Transcript; Transcriptional Regulation; Untranslated RNA; Volition; Whole-Cell Recordings; Work; addiction; cell type; circular RNA; cocaine self-administration; cocaine use; drug of abuse; human subject; human tissue; in vivo; innovation; insight; knock-down; mouse model; novel; opioid abuse; response; reward circuitry; transcriptome sequencing; transcriptomics; virtual

PROJECT SUMMARY/ABSTRACT– PROJECT 2 Project 2's objective is to understand the role for circular RNAs (circRNAs) in the mechanisms by which cocaine remodels the dorsal striatum (DS) and nucleus accumbens (NAc) to precipitate compulsive cocaine- seeking behaviors. circRNAs are highly conserved single-stranded ‘back-spliced’ RNAs in which the 5′ and 3′ ends of the transcript are covalently joined. Emerging evidence suggests that circRNAs are a major class of regulatory noncoding RNAs that are enriched in brain and that play key roles in basic aspects of neuronal function, but their involvement in the molecular, cellular, and behavioral actions of cocaine (and other drugs of abuse) has yet not been investigated. We will characterize patterns of circRNA expression in DS and NAc of mice that show compulsive-like cocaine consumption using paired-end ribominus RNA-sequencing. We will also assess circRNA expression in postmortem striatal tissues from humans with cocaine use disorders. We already have robust evidence for prominent cocaine regulation of several circRNAs in these brain regions. Those cocaine-responsive circRNAs that show similar abnormal expression in mice and humans will be prioritized for further investigation. We will determine whether prioritized cocaine-responsive circRNAs are regulated specifically in different types of DS and NAc neurons. We will then investigate the role played by these prioritized cocaine-responsive circRNAs in regulating the molecular and cellular responses to cocaine within these cell types of DS and NAc. This will be accomplished by use of in vivo CRISPR technology or RNAi to knockdown prioritized circRNAs in a cell type-specific manner and assess the influence of these circrRNAs in controlling baseline and cocaine-induced alterations in the intrinsic excitability of the neurons as well as their transcriptional responses to cocaine. Finally, we will investigate the ways in which cocaine-responsive circRNAs control compulsive-like cocaine self-administration behavior including relapse in mice. These highly innovative studies promise to yield fundamentally new insights into the molecular and cellular mechanisms of cocaine addiction, with parallel future studies planned for opiate drugs of abuse.

项目总结/摘要-项目2 项目2的目标是了解环状RNA（circRNA）在以下机制中的作用： 可卡因重塑背侧纹状体（DS）和背侧核（NAc），以沉淀可卡因强迫性。 寻求行为。circRNA是高度保守的单链“反向剪接”RNA，其中5′和3′端的 转录物的末端共价连接。新出现的证据表明，circRNA是一个主要类别的 在大脑中富集的调节性非编码RNA，在神经元的基本方面发挥关键作用， 功能，但它们参与可卡因（和其他药物）的分子，细胞和行为作用。 （滥用）尚未得到调查。我们将描述DS和NAc中circRNA表达的模式， 使用配对末端ribominus RNA测序显示出强迫性可卡因消耗的小鼠。我们将 还评估了来自可卡因使用障碍的人的死后纹状体组织中的circRNA表达。我们 已经有了强有力的证据证明可卡因对这些大脑区域中的几种circRNA有显著的调节作用。 那些在小鼠和人类中表现出类似异常表达的可卡因反应性circRNA将被发现。 优先进行进一步调查。我们将确定优先可卡因反应的circRNA是否 在不同类型的DS和NAc神经元中特异性调节。然后我们将调查所扮演的角色， 这些优先可卡因反应的circRNA在调节分子和细胞对可卡因的反应， 在这些细胞类型的DS和NAc中。这将通过使用体内CRISPR技术或RNAi技术来实现。 以细胞类型特异性方式敲低优先的circRNA，并评估这些circRNA的影响， 在控制基线和可卡因诱导的神经元内在兴奋性的改变以及它们的 对可卡因的转录反应最后，我们将研究可卡因反应的方式， circRNA控制小鼠中包括复发在内的强迫样可卡因自我给药行为。这些高度 创新的研究有望从根本上产生新的见解的分子和细胞机制， 可卡因成瘾，计划今后对阿片类药物滥用进行平行研究。