Circular RNA signaling in opioid seeking phenotypes

摘要： 物质使用障碍是一种代价高昂、使人衰弱的慢性疾病，其特点是复发率高 尽管成功地尝试了康复和禁欲。此外，药物过量和 与阿片类药物相关的死亡在过去十年中急剧上升， 公共卫生负担重。为了确定治疗阿片类药物使用障碍的假定药物靶点， 必须阐明控制阿片寻求的分子机制。该提案的目的是解决 通过探索全新的机制， 调节阿片类药物从一类称为环状RNA的RNA中寻找。尽管环状RNA从未被 在药物寻求行为的研究中，这种RNA有可能广泛影响细胞信号传导 在表观遗传，转录和翻译水平，这反过来又可能影响广泛的行为， 表型环状RNA是单链转录剪接产物，其产生自RNA的反向剪接。 mRNA的3'至5'末端。这种剪接产物开始在大脑中被探索，其中一些在大脑中表达。 比它们的mRNA对应物更高的水平，并且据报道调节关键过程，包括 microRNA活性，基因表达，蛋白质翻译，甚至认知行为。我们发现了一组 大鼠海洛因自我给药后调节的环状RNA及其生物合成酶 眶额皮层（OFC），一个对奖赏寻求至关重要的大脑区域。在颁奖期间，我们将探讨 OFC环状RNA信号通路的改变有助于阿片样物质寻求表型的假说。我们将使用一个 多学科方法来检查海洛因诱导的循环的细胞和行为后果 阿片样物质寻求大鼠模型中的RNA表达。首先，海洛因相关的环状RNA将在基因上 在OFC中操纵，以确定它们对阿片类药物的动机和阿片类药物寻求复发的贡献 在灭绝或禁欲之后。然后，我们将定义海洛因相关环状RNA的细胞表型， OFC旨在深入了解海洛因改变了circRNA生物合成的细胞回路 exposure.最后，我们将确定海洛因相关的环状RNA下游的信号通路， OFC来描述这种新型RNA在药物暴露神经元内的分子相互作用。我们 海洛因相关环状RNA的数据集以前从未在大脑中进行过研究， 药物滥用领域的全新研究领域。因此，前所未有的结果 拟议中的研究将揭示控制药物寻求的新机制，并提供必要的见解 药物滥用的神经生物学

Abstract: Substance use disorder is a costly and debilitating chronic disorder, characterized by high rates of relapse despite successful attempts at rehabilitation and periods of abstinence. In addition, the rates of overdose and death associated with the opioid class of drugs have dramatically risen over the last decade and represent a major public health burden. To identify putative drug targets for the treatment of opioid use disorder, the molecular mechanisms that govern opioid seeking must be delineated. The goal of this proposal is to address this critical barrier in the field of substance abuse research by exploring entirely new mechanisms that modulate opioid seeking from a class of RNA called circular RNAs. Although circular RNAs have never been studied in drug seeking behaviors, this species of RNA has the potential to broadly influence cellular signaling at the epigenetic, transcriptional and translational levels, which in turn may impact a wide range of behavioral phenotypes. Circular RNAs are single-stranded transcriptional splice products that result from backsplicing of 3' to 5' ends of mRNA. Such splice products are beginning to be explored in the brain, with some expressed at higher levels than their mRNA counterparts, and have been reported to modulate critical processes including microRNA activity, gene expression, protein translation and even cognitive behaviors. We have identified a set of circular RNAs and their biogenesis enzymes that are regulated after heroin self-administration in the rat orbitofrontal cortex (OFC), a brain region essential to reward seeking. During the award period, we will explore the hypothesis that altered OFC circular RNA signaling contributes to opioid seeking phenotypes. We will use a multi-disciplinary approach to examine the cellular and behavioral consequences of heroin-induced circular RNA expression in a rat model of opioid seeking. First, heroin-associated circular RNAs will be genetically manipulated in the OFC to determine their contribution to motivation for opioids and relapse to opioid seeking after extinction or abstinence. We will then define the cellular phenotype of heroin-associated circular RNAs in the OFC for insights into the cellular circuits that have altered circRNA biogenesis as a result of heroin exposure. Finally, we will identify signaling pathways downstream of heroin-associated circular RNAs in the OFC to describe the molecular interactions of this novel class of RNA within drug-exposed neurons. Our dataset of heroin-associated circular RNAs have never been studied in the brain before and represent an entirely new area of research in the field of substance abuse. Therefore, the outcome of the unprecedented proposed studies will shed light on novel mechanisms that govern drug seeking and provide essential insight into the neurobiology of substance abuse.