Circular RNA signaling in opioid seeking phenotypes

摘要： 物质使用障碍是一种代价高昂且使人虚弱的慢性疾病，其特征是复发率高。 尽管成功地尝试了康复和禁欲时期。此外，服药过量和吸毒的比率 与阿片类药物相关的死亡人数在过去十年里急剧上升，代表着一种 重大公共卫生负担。为了确定治疗阿片类药物使用障碍的可能药物靶点， 必须描述管理阿片类药物寻找的分子机制。这项提案的目标是解决 通过探索全新的机制，这一物质滥用研究领域的关键障碍 调节从一类被称为环状RNA的RNA寻找阿片类药物。尽管环状RNA从来没有被 在药物寻找行为的研究中，这种rna具有广泛影响细胞信号的潜力。 在表观遗传、转录和翻译水平上，这反过来可能影响广泛的行为 表型。环状RNA是单链转录剪接产物，由 信使核糖核酸3‘端至5’端。这种剪接产物正开始在大脑中被探索，其中一些表达于 比其对应的信使核糖核酸水平更高，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。我们已经确定了一套 海洛因自身给药对环状RNA及其生物发生酶调控的影响 眼眶前额叶皮质(OFC)，这是大脑中对寻求奖励至关重要的区域。在颁奖期间，我们将探索 OFC环状RNA信号改变与阿片类药物寻找表型有关的假说。我们将使用 多学科方法研究海洛因诱发循环的细胞和行为后果 大鼠阿片类药物寻找模型中RNA的表达。首先，与海洛因相关的环状RNA将是基因上的 在OFC中被操纵，以确定他们对阿片类药物的动机和对阿片类药物寻求的复发的贡献 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 对海洛因引起的CircRNA生物发生改变的细胞电路的洞察 曝光。最后，我们将确定海洛因相关环状RNA下游的信号通路 OFC来描述这类新的RNA在药物暴露的神经元中的分子相互作用。我们的 与海洛因相关的环状RNA数据集以前从未在大脑中被研究过，并且代表着一种 药物滥用领域的全新研究领域。因此，结果是史无前例的 拟议的研究将阐明管理药物寻找的新机制，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.