Making and breaking opioid memories to prevent relapse

项目摘要 反复接触毒品会导致广泛的细胞变化，表现为适应不良行为。 还有毒瘾。这些行为变化可能会使人虚弱，而上瘾的病理通常是终生的 痛苦。事实上，即使在多年的禁欲之后，接触与毒品有关的药物也可能导致复发。 暗示。这是毒品成瘾的一个普遍特征，并存在于毒品类别中，但我们缺乏一个明确的 理解这些变化为什么会如此持久。许多类别的蛋白质调节失调 与线索诱发的复发有牵连，但复发的倾向在这些人的半衰期过后仍然存在 蛋白质，这表明上游表观遗传变化是允许转录格局的 会产生这些行为异常。理解最终产生这些的表观遗传变化 细胞变化将极大地扩大潜在治疗干预的靶点数量。至 要了解线索诱发复发的表观遗传机制，我们必须确定 被激活以驱动寻找行为的细胞。我们的初步数据显示，在任何给定的大脑区域，只有 一小部分细胞对给定的刺激被激活--这组被激活的神经元被称为神经 合唱团。因此，理解大脑的下一个前沿将是准确地定义哪些细胞被激活， 它们何时被激活，以及为什么。在这里，我们结合了一些技术，允许我们记录、处理和 阿片类药物自我给药和随后的线索触发药物寻找过程中的序列神经系综 确定每个神经元群体中的转录活动如何决定哪些细胞被激活。通过 结合单细胞线索诱导寻找过程中的体内细胞分辨钙成像 测序--在相同的动物中--我们将定义控制神经活动的转录网络 驱使人们寻找毒品的模式。接下来，使用表观遗传学方法和CRISPR/dCas9融合构建，我们 将在神经元中选择性地定义和操纵活动反应基因的表观遗传图景 被与毒品有关的线索激活。这项提议将使我们能够定义准确的神经系综来指导 药物寻找以及这些神经元内的转录网络如何控制引导 行为。通过定义这些机制，这个奖项将允许进行研究，推动我们如何 了解大脑中的信息编码，并扩展我们对如何 操纵这些过程，以减少跨药物类别的复发。

Project Summary Repeated drug exposure produces widespread cellular alterations that can manifest in maladaptive behaviors and addiction. These behavioral alterations can be debilitating, and the pathology of addiction is often a life-long affliction. Indeed, even after years of abstinence, relapse can be precipitated by exposure to drug-associated cues. This is a ubiquitous property of drug addiction, and is present across drug class, yet we lack a clear understanding of how these changes can be so long lasting. Dysregulation of many classes of proteins have been implicated in cue-evoked relapse, yet the propensity to relapse persists well past the half-life of these proteins, suggesting that upstream epigenetic changes are permissive to the transcriptional landscape that produces these behavioral aberrations. Understanding the epigenetic alterations that ultimately produce these cellular changes will great expand the number of targets available for potential therapeutic interventions. To approach understanding the epigenetic mechanisms that underlie cue-induced relapse, we must identify the cells that are activated to drive seeking behavior. Our preliminary data show that in any given brain region, only a small percentage of cells are activated to a given stimulus – this group of activated neurons is termed a neural ensemble. Thus, the next frontier of understanding the brain will be defining exactly which cells are activated, when they are activated, and why. Here we combine techniques that allow us to record, manipulate, and sequence neural ensembles during opioid self-administration and subsequent cue-triggered drug seeking to determine how transcriptional activity within each neuronal population dictates which cells are activated. By combining in vivo cellular resolution calcium imaging during cue-induced seeking followed by single cell sequencing - in the same animals - we will define the transcriptional networks that control the neural activity patterns that drive drug seeking. Next, using epigenetic approaches and CRISPR/dCas9 fusion constructs, we will define and manipulate the epigenetic landscape at activity-responsive genes selectively in neurons that are activated by drug-associated cues. This proposal will allow us to define the precise neural ensembles that guide drug seeking and how transcriptional networks within these neurons control the neural activity profiles that guide behavior. By defining these mechanisms, this award will allow research that pushes the boundaries of how we approach understanding information encoding in the brain and expand our understanding of how we can manipulate these processes to reduce relapse across drug classes.