A comprehensive dissection of cell types, circuits and molecular adaptations during opioid use

对阿片类药物使用过程中的细胞类型、回路和分子适应的全面剖析

基本信息

批准号：
10598151
负责人：
MARK J SCHNITZER
金额：
$ 139.99万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-06-01 至 2026-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10598151
关键词：
Abstinence Adenylate Cyclase Amygdaloid structure Anatomy Architecture Arrestins BRAIN initiative Behavioral Binding Brain Brain imaging Brain region Catalogs Cells Censuses Chronic Complex Computer Analysis Data Data Coordinating Center Data Set Databases Dependence Dissection Electrophysiology (science)Ensure Foundations Functional Imaging Funding G-Protein-Coupled Receptors GTP-Binding Proteins Gene Expression Gene Family Genes Habenula Hippocampus Hypothalamic structure Immunohistochemistry In Situ Hybridization Ion Channel Knowledge Label Length Link Maps Mediating Modeling Molecular Molecular Target Morphine Mus National Institute of Drug Abuse Nerve Tissue Nervous System Neurobiology Neurons Nucleus Accumbens Nucleus solitarius ORL1 receptor Opiate Addiction Opioid Opioid Analgesics Opioid Peptide Opioid Receptor Optics Output Pain Pain management Pattern Phosphotransferases Postoperative Pain Prefrontal Cortex Protein Isoforms Protein Subunits Protocols documentation Reporter Research Research Personnel Resolution Resources Science Self Administration Slice Specificity Surveys Synapses System Technology Thalamic structure Toxic effect Ventilatory Depression Ventral Tegmental Area Viral Withdrawal Work addiction brain cell cell type clinically relevant computer studies conditioned place preference cranium dorsal raphe nucleus endogenous opioids experimental group gene network imaging modality imaging study improved in vivo innovation innovative technologies midbrain central gray substance molecular dynamics molecular sequence database mouse model mu opioid receptors neural neural circuit nociceptin novel novel therapeutic intervention opioid exposure opioid use opioid use disorder parabrachial nucleus preBotzinger complex prevent radioligand receptor response side effect single nucleus RNA-sequencing single-cell RNA sequencing transcriptomics

项目摘要

PROJECT SUMMARY A major barrier in the field of opioid research is our limited understanding of the organization of the opioid system in the brain: we still do not know which cell types express each opioid receptor (mu, delta, kappa, nociceptin) to mediate the effects of endogenous and exogenous opioids, nor what other molecules are present in these cells. Without this knowledge, understanding how opioids alter activity in circuits to produce behavioral effects remains elusive. This gap in knowledge prevents the identification of molecular targets to potentiate opioid analgesia and mitigate the deleterious effects of opioid use disorder (OUD), including addiction and respiratory depression. To fill this gap in knowledge, we propose to leverage the uniquely massive single-cell RNA sequencing (scRNA-seq) database generated by the Allen Institute for Brain Science (>3.4 million cells throughout the entire mouse brain) as part of the BRAIN Initiative Cell Census Network (BICCN) effort. Using this database, we will establish a comprehensive catalog of all the cell types that express each opioid receptor and peptide throughout the brain, as well as the co-expression of gene networks that mediate or regulate opioid actions, including other G protein-coupled receptors and cellular effectors of opioid receptors (Aim 1). Further, we will use our well- established high-throughput single-cell RNA-seq pipelines to characterize the cell-type-specific molecular adaptations that occur during chronic opioid exposure, withdrawal, and abstinence, using a clinically relevant model of post-surgical pain for which opioids are typically prescribed (Aim 2). Finally, we will leverage this novel knowledge to dissect the mechanisms of action of opioids by performing advanced circuit mapping and in vivo functional imaging studies of cell types expressing opioid receptors in the prefrontal cortex (PFC), a region critical to both opioid analgesia and addiction (Aim 3). Our transformative work will be the first to combine several highly innovative technologies at the molecular, circuit, and neural ensemble levels, including high-throughput scRNA-seq, new viral strains with improved transsynaptic transfer and decreased toxicity for circuit mapping, the crystal skull and miniscope- microprism optical approaches for in vivo wide-field imaging of brain state and for recording dynamics of molecularly defined neuron types in freely moving mice undergoing opioid analgesia and addiction paradigms. We have an extraordinary interdisciplinary team of investigators with highly complementary expertise in the neurobiology of opioids and the distribution and function of their receptors in pain and addiction circuits, the molecular and anatomical brain architecture, large-scale cell type characterization and circuit mapping, and highly innovative brain imaging methods as applied to the study of neural circuits. Overall, this research aims to generate an exceptional resource for the opioid field (to be made publicly available through the NIDA-funded SCORCH data coordination center) to explain how opioids change the brain, and discover novel therapeutic approaches to prevent and treat OUD.

项目概要阿片类药物研究领域的一个主要障碍是我们对阿片类药物系统的组织了解有限在大脑中：我们仍然不知道哪些细胞类型表达每种阿片受体（mu、delta、kappa、痛敏素）介导内源性和外源性阿片类药物的作用，也介导这些细胞中存在的其他分子的作用。如果没有这些知识，我们仍然无法了解阿片类药物如何改变回路活动以产生行为效应难以捉摸。这种知识差距阻碍了增强阿片类药物镇痛的分子靶标的识别和减轻阿片类药物使用障碍 (OUD) 的有害影响，包括成瘾和呼吸抑制。为了填补这一知识空白，我们建议利用独特的大规模单细胞 RNA 测序 (scRNA-seq) 数据库由艾伦脑科学研究所生成（整个过程中超过 340 万个细胞）小鼠大脑）作为 BRAIN Initiative 细胞普查网络 (BICCN) 工作的一部分。使用这个数据库，我们将建立表达每种阿片受体和肽的所有细胞类型的综合目录大脑，以及介导或调节阿片类药物作用的基因网络的共表达，包括其他 G 蛋白偶联受体和阿片受体的细胞效应器（目标 1）。此外，我们将利用我们的良好建立了高通量单细胞 RNA-seq 管道来表征细胞类型特异性分子使用临床相关的方法，在长期阿片类药物暴露、戒断和戒断期间发生的适应通常使用阿片类药物治疗的术后疼痛模型（目标 2）。最后，我们将利用这本小说通过执行高级电路图谱和体内分析来剖析阿片类药物作用机制的知识对前额皮质 (PFC) 中表达阿片受体的细胞类型进行功能成像研究，该区域是一个关键区域阿片类药物镇痛和成瘾（目标 3）。我们的变革性工作将是第一个将多项高度创新技术结合起来的工作分子、电路和神经整体水平，包括高通量 scRNA-seq、新病毒株改善突触传递并降低电路映射、水晶头骨和微型显微镜的毒性用于大脑状态体内广域成像和记录大脑状态动态的微棱镜光学方法接受阿片类镇痛和成瘾范例的自由活动小鼠的分子定义神经元类型。我们拥有一支出色的跨学科研究团队，他们在以下领域具有高度互补的专业知识阿片类药物的神经生物学及其受体在疼痛和成瘾回路中的分布和功能，分子和解剖学脑结构、大规模细胞类型表征和电路映射，以及高度创新的脑成像方法应用于神经回路的研究。总体而言，这项研究旨在为阿片类药物领域提供特殊的资源（将公开）可通过 NIDA 资助的 SCORCH 数据协调中心获得）来解释阿片类药物如何改变大脑，并发现预防和治疗 OUD 的新治疗方法。