Genomic profiling mediating the protective effect of social reward on opioid craving

项目摘要 目前的阿片类药物危机是一个重大的公共卫生问题，尽管有食物和药品供应， 药物管理局批准的治疗（美沙酮，丁丙诺啡和纳洛酮）。一项至关重要的挑战是 在接受治疗的患者中发生失误或复发，特别是考虑到复发有过量的风险。 因此，存在未满足的需要，以推进可以克服药物治疗的局限性的新的替代干预。 阿片类药物使用障碍（OUD）的现有治疗方案。成功有限的原因是 OUD的治疗是复杂和多因素的。一个潜在的局限性可以归因于经典的 OUD的动物模型，很少包含社会因素。我们最近开发了一种操作性大鼠模型， 海洛因和社会交往之间的选择，并显示：（1）大鼠强烈偏好操作性社会交往 （2）社会选择诱导的戒断（自愿戒断）阻止了海洛因寻求 (in一种药物成瘾的模型）。调节社会保护作用的遗传和电路机制 海洛因成瘾的奖励是未知的。这项提案的首要目标是研究基因组图谱， 有助于社会互动对海洛因渴望的缓冲作用。我们将集中在中央核 杏仁核（CeA）及其预测的基础上，我们的研究结果的作用，CeA在孵化 社会选择诱导的自愿戒断后的精神兴奋剂渴求及其作用的研究 强迫戒毒后对毒品的渴望。在目标1中，我们将生成单个- 来自CeA的核RNA-seq及其在大鼠中的投射（使用顺行跨突触病毒）， 孵化的测试后，无论是基于社会的自愿或强迫禁欲。另外。我们将解读 来自BRAIN的每个脑区域中的转录组细胞类型的详细本体内的转录组数据 Initiative Cell Census Network，以及人类死后杏仁核和杏仁核的snRNA-seq 来自HIV背景下单细胞阿片样物质反应的阿片样物质使用障碍供体（SCORCH） 财团目的2：利用腺相关病毒逆行标记CeA投射神经元细胞核 并将转录组数据与来自大型联合体的互补转录组数据集整合。在Aim中 3、我们将联合收割机结合我们的实验相关的社会选择动物模型与先进的CRISPRa， CRISPRi AAV工具操纵CeA或投射区中关键枢纽基因的转录， 表明它们在基于社会的对海洛因渴求的保护中的因果作用。我们的建议将提供新的 社会奖励对海洛因保护作用的基因组分析及其机制 在一个新的基于社会的老鼠模型中的渴望。这些结果将提高我们的机械理解的作用 的社会互动。

Project Summary The current opioid crisis is a major public health problem, and it persists despite the availability of Food and Drug Administration–approved treatments (methadone, buprenorphine, and naltrexone). A critical challenge is the occurrence of lapses or relapses in treated patients, especially given that relapses carry a risk of overdose. Therefore, there is an unmet need to advance novel alternative interventions that may overcome limitations of currently available treatment options for opioid use disorders (OUDs). The reason for the limited success of treatments for OUDs are complex and multifactorial. One potential limitation could be attributed to classical animal model of OUDs which rarely incorporate social factors. We recently developed an operant rat model of choice between heroin and social interaction and showed that: (1) rats strongly prefer operant social interaction over heroin, and (2) social choice-induced abstinence (voluntary abstinence) prevents heroin seeking over time (in a model of drug craving). The genetic and circuit mechanisms mediating the protective effect of social reward on heroin craving are unknown. The overarching goal of this proposal is to study the genomic profiling contributing to the buffering effect of social interaction on heroin craving. We will focus on the central nucleus of the amygdala (CeA) and its projections based on our findings on the role of CeA in incubation of psychostimulant craving after social choice-induced voluntary abstinence and previous studies on its role in incubation of drug craving after forced abstinence across drug classes. In Aim 1, we will generate single- nucleus RNA-seq from CeA and its projections (using anterograde transynaptic viruses) in rats during the incubation’s tests after either social-based voluntary or forced abstinence. Additionally. we will interpret the transcriptomic data within detailed ontologies of transcriptomic cell types in each brain region from the BRAIN Initiative Cell Census Network, as well as with snRNA-seq of post-mortem amygdala and insula of human donors with opioid use disorder from the Single-Cell Opioid Responses in the Context of HIV (SCORCH) consortium. In Aim 2, we will use retrograde adeno-associated viruses to label CeA projection neuron nuclei and integrate the transcriptomic data with complementary transcriptomic datasets from large consortia. In Aim 3, we will combine our translationally relevant social choice animal model with advanced CRISPRa and CRISPRi AAV tools to manipulate transcription of key hub genes in either CeA or projection regions to demonstrate their causal role on social-based protection of heroin craving. Our proposal will provide new insights into genomic profiling and its mechanisms underlying the protective effect of social reward on heroin craving in a novel social-based rat model. These results will improve our mechanistic understanding of the role of social interaction in OUDs.