Integrative single-cell spatial transcriptomic, anatomical, and functional profiling of brain-wide ensembles engaged by opioid relapse

与阿片类药物复发有关的全脑整体的综合单细胞空间转录组、解剖学和功能分析

• 批准号: 10772455
• 负责人: Sam Golden
• 金额: $ 80.15万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-30 至 2028-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10772455
• 关键词: Address; Anatomy; Atlases; Bar Codes; Behavior; Behavioral; Biological Assay; Brain; Brain Mapping; Brain region; Cells; Classification; Clinical; Communities; Coupling; Cues; Data; Data Set; Drug Exposure; Electrophysiology (science); Emotional; Euphoria; FOS gene; Female; Fentanyl; Future; Gene Expression; Gene Expression Profile; Genes; Genetic; Genetic Transcription; Goals; Heroin; Image; Immediate-Early Genes; Implant; Individual; Individual Differences; Infusion procedures; Intravenous; Machine Learning; Maps; Methodology; Methods; Modeling; Molecular; Morphine; Mus; Neurons; Neurosciences; Opioid; Oral; Oral Characters; Pharmaceutical Preparations; Phenotype; Population; Process; Relapse; Reporting; Resolution; Risk; Route; Self Administration; Slice; Specificity; Technology; Time; United States; Viral; Vulnerable Populations; Withdrawal; brain cell; brain tissue; brain volume; cell type; combat; differential expression; drug seeking behavior; fentanyl abuse; fentanyl seeking; fentanyl self-administration; fentanyl use; functional adaptation; gene conservation; in vivo; individual variation; innovation; interest; learned behavior; male; multimodal data; multimodality; neural; neural circuit; novel; open source; opioid abuse; opioid mortality; opioid use; recruit; relapse risk; response; spatial integration; synthetic opioid; technology platform; transcriptome; transcriptomic profiling; transcriptomics

PROJECT SUMMARY Opioid abuse is devastating communities across the United States and is responsible for untold suffering. The synthetic opioid fentanyl, whether due to prescription or illicit use, is involved in nearly half of reported opioid- related deaths. Unlike heroin and morphine, fentanyl is commonly administered through non-intravenous routes, but we have a limited mechanistic understanding of fentanyl use and abuse vulnerability. A leading hypothesis for the transition from use to abuse, in vulnerable individuals, is the recruitment of drug-induced gene expression changes in certain brain circuits and cells following repeated drug exposure. However, identifying such circuit- and cell-type specific populations, in combination with their underlying genetic and functional adaptations, is often limited by the resolution, throughput, and data registration of current assays. The overall goal of this project is to (i) generate machine--guided behavioral characterization of oral and intravenous fentanyl self-administration and reinstatement in male and female mice that captures individual risk vulnerability, and (ii) perform brain-wide mapping by integrating new enabling technological platforms including single-cell spatial transcriptomics (Pixel- seq), whole brain cell type specific circuit connectivity, and functional distributed brain-wide neural activity recordings (Neuropixel 2.0), recently developed by PIs in our team. To realize the potential for the molecular and functional brain mapping and data registration, we have three Specific Aims: 1) perform single-cell spatial transcriptomic profiling, with circuit-specificity, in mice with varying degrees of fentanyl-seeking behavior; 2) perform single-cell whole-brain activity mapping, with cell-type and circuit-specificity, in mice with varying degrees of fentanyl-seeking behavior; 3) perform brain-wide distributed large-scale electrophysiological recordings during oral fentanyl-seeking and integrate spatially resolved transcriptomic data with recording data. In the first Aim, Pixel-seq will be used to generate cell atlases, analyze drug- and behavior-associated spatially conserved gene expression, and map neuronal connectivity by coupling with retrograde viral tracing. In the second Aim, we will perform immediate early gene-based whole-brain activity mapping, contextualized by cell type-specific afferent connectivity, after fentanyl self-administration and reinstatement. In the third Aim, we will first perform Neuropixel2.0 electrophysiological recording of the brain-wide distributed regions of interest, and then integrate and align Neuropixel2.0 and Pixel-seq data in a new assay called NeuroPixel-seq (NP-seq). The proposed project is innovative as our integrative approaches will, for the first time, generate spatial multimodal data of unprecedented depth and resolution within the context of opioid relapse risk. It is significant because our data will provide a much-needed accessible oral fentanyl self-administration model for the neuroscience community, paired with the first fentanyl cellular-resolution atlas, allowing non-specialized labs an accessible beachhead for participating in the identification of the mechanistic basis for fentanyl use and relapse in mice.

项目概要 阿片类药物的滥用正在给美国各地的社区带来毁灭性的打击，并造成难以言喻的痛苦。这 合成阿片类药物芬太尼，无论是处方使用还是非法使用，已报告的阿片类药物中近一半涉及合成阿片类药物芬太尼。 相关死亡。与海洛因和吗啡不同，芬太尼通常通过非静脉途径给药， 但我们对芬太尼的使用和滥用脆弱性的机制了解有限。一个主要假设 在易受伤害的个体中，从使用到滥用的转变是招募药物诱导的基因表达 反复接触药物后某些脑回路和细胞会发生变化。然而，识别这样的电路 和细胞类型特定群体，结合其潜在的遗传和功能适应， 通常受到当前检测的分辨率、通量和数据记录的限制。本项目的总体目标 (i) 生成口服和静脉注射芬太尼自我给药的机器引导行为特征 以及在雄性和雌性小鼠中进行恢复，以捕获个体风险脆弱性，并且（ii）在全脑范围内进行 通过集成新的使能技术平台（包括单细胞空间转录组学（Pixel- seq）、全脑细胞类型特定电路连接和功能性分布式全脑神经活动 录音 (Neuropixel 2.0)，最近由我们团队的 PI 开发。实现分子和 功能性大脑图谱和数据注册，我们有三个具体目标：1）执行单细胞空间 对具有不同程度的芬太尼寻求行为的小鼠进行具有电路特异性的转录组分析； 2） 在具有不同细胞类型和电路特异性的小鼠中进行单细胞全脑活动图谱 寻求芬太尼行为的程度； 3）进行全脑分布式大规模电生理 口服芬太尼寻求过程中的记录，并将空间解析的转录组数据与记录数据整合。 在第一个目标中，Pixel-seq 将用于生成细胞图谱，分析与药物和行为相关的空间 保守的基因表达，并通过与逆行病毒追踪相结合来绘制神经元连接图。在 第二个目标，我们将立即进行基于基因的早期全脑活动图谱，以细胞为背景 芬太尼自我给药和恢复后，类型特异性传入连接。在第三个目标中，我们将 首先对全脑分布的感兴趣区域进行Neuropixel2.0电生理记录，以及 然后在名为 NeuroPixel-seq (NP-seq) 的新测定中整合并对齐 Neuropixel2.0 和 Pixel-seq 数据。这 拟议的项目具有创新性，因为我们的综合方法将首次产生空间多模式 在阿片类药物复发风险的背景下，数据具有前所未有的深度和分辨率。这很重要，因为我们 数据将为神经科学提供急需的口服芬太尼自我给药模型 社区，与第一个芬太尼细胞分辨率图集配对，使非专业实验室能够访问 参与确定小鼠芬太尼使用和复发的机制基础。

项目成果

Individual Differences in Volitional Social Self-Administration and Motivation in Male and Female Mice Following Social Stress.

社会压力后雄性和雌性小鼠的自愿社会自我管理和动机的个体差异。

• 发表时间: 2024-01-18
• 影响因子: 10.6
• 作者: Navarrete, Jovana;Schneider, Kevin N;Smith, Briana M;Goodwin, Nastacia L;Zhang, Yizhe Y;Salazar, Axelle S;Gonzalez, Yahir E;Anumolu, Pranav;Gross, Ethan;Tsai, Valerie S;Heshmati, Mitra;Golden, Sam A
• 通讯作者: Golden, Sam A