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: 10533296
• 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 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10533296
• 关键词: Abstinence; Autopsy; Aversive Stimulus; Awareness; 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; Modeling; 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; Regulation; Relapse; Role; Self Administration; Site; Slice; Substance Use Disorder; Synapses; Synaptic Transmission; Transcript; Transcriptional Regulation; Untranslated RNA; Whole-Cell Recordings; Work; addiction; cell type; circular RNA; cocaine seeking; cocaine self-administration; cocaine use; drug of abuse; human subject; human tissue; in vivo; innovation; insight; knock-down; mouse model; neuronal excitability; 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.

项目摘要/摘要--项目2 项目2：S的目标是了解环状RNA(CircRNA)在 可卡因重塑背侧纹状体(DS)和伏隔核(NAC)以沉淀强迫性可卡因- 寻找行为。CircRNAs是高度保守的单链‘后剪接’RNA，其中5‘和3’ 转录本的末端是共价连接的。新出现的证据表明，CircRNA是一类主要的 富含于大脑并在神经元的基本方面发挥关键作用的调节性非编码RNA 功能，但它们参与可卡因(和其他毒品)的分子、细胞和行为行为 虐待)尚未得到调查。我们将表征CircRNA在DS和NAC中的表达模式 使用成对末端核糖核酸测序显示出强迫性可卡因消费的小鼠。我们会 还评估了可卡因使用障碍患者死后纹状体组织中CircRNA的表达。我们 已经有强有力的证据表明，可卡因对这些大脑区域中的几个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CircRNA将是 被列为进一步调查的优先事项。我们将确定优先处理的可卡因反应CircRNA是否 在不同类型的DS和NAC神经元中特异调节。然后我们将调查他所扮演的角色 这些优先反应可卡因的CircRNA在调节可卡因的分子和细胞反应中 在DS和NAC这两种细胞类型中。这将通过使用体内CRISPR技术或RNAi来完成 以特定细胞类型的方式敲除优先考虑的CircRNA并评估这些CircRNAs的影响 在控制基线和可卡因诱导的神经元内在兴奋性以及它们的 可卡因的转录反应。最后，我们将调查可卡因反应的方式 CircRNAs控制强迫性类可卡因的自我给药行为，包括小鼠的复发。这些高度 创新性的研究有望从根本上深入了解人类免疫缺陷的分子和细胞机制。 可卡因成瘾，同时计划对鸦片类药物滥用进行平行的未来研究。