Mapping brain-wide opioid actions by profiling neuronal activities and in vivo cellular target engagement

通过分析神经元活动和体内细胞靶标参与来绘制全脑阿片类药物作用

• 批准号: 10775623
• 负责人: Laura M. Bohn
• 金额: $ 83.99万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 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/10775623
• 关键词: Affect; Affinity; Agonist; Architecture; Area; Atlases; Behavior; Binding; Binding Sites; Body part; Brain; Brain Mapping; Brain imaging; Brain region; Buprenorphine; Cells; Chemical Structure; Chemicals; Chemistry; Coupled; DNA; Data; Drug Targeting; FOS gene; Genes; Goals; Heterogeneity; Image; In Situ Hybridization; In Vitro; Individual; Ligands; Light; Link; Maps; Measures; Mediating; Molecular; Mus; Neurons; Opioid; Opioid Receptor; Overdose; Pharmaceutical Preparations; Pharmacology; Phenotype; Population; Proteins; Public Health; RNA; Research Personnel; Resolution; Salvia; Signal Transduction; Site; Somatotype; Specificity; Stains; Structure-Activity Relationship; Synthesis Chemistry; System; Techniques; Testing; Tissues; Validation; Visualization; Work; activity marker; addiction liability; biomarker identification; brain cell; cell type; cellular targeting; design; endogenous opioids; imaging approach; in situ imaging; in vivo; lead optimization; multimodality; neural; neuroregulation; new technology; off-target site; opioid use disorder; pain reduction; pharmacologic; preference; receptor; salvinorin A; screening; side effect; transcriptome; transcriptomics

Abstract Opioids are highly effective at reducing pain, but their potential for addiction and overdose has led to a growing public health crisis. Researchers have attempted to develop new opioid compounds that are less likely to be abused and have fewer side effects, but these efforts have been difficult. The endogenous opioid system has multiple receptors and ligands heterogeneously expressed across different parts of the body and cell types. Tremendous work has been done to delineate the relationships between opioid receptors (ORs) and ligands. However, the specificity of ligand-receptor engagement often depends on relative affinities at predetermined targets. In general, in vivo spatial and cellular heterogeneity of the brain obscure opioid actions, making them hard to predict based on receptor affinity alone. Currently, single-cell and spatial transcriptomics are transforming our understanding of brain architecture. However, there is a significant gap in how we measure opioid actions and align them with the spatially resolved cellular atlas of the brain. Levering emerging CATCH and inverse activity marker (IAM) techniques, we propose multimodal profiling of opioid actions with spatial and single-cell resolution across the entire mouse brain. Using three pharmacologically diverse opioids, we aim to map neuronal activities and cellular binding of these drugs onto the entire mouse brain in an unbiased way and register them with cell types identified from single-cell transcriptomics. Furthermore, we will test whether a drug’s affinities across different ORs determine its in vivo cell and neural ensemble engagement. Not only would this project provide a circuit-level mechanism linking the molecular pharmacology to brain-wide opioid actions, but also lay out a roadmap for evaluating and developing new opioids, e.g., by incorporating regional and cell-type preference into the structure-activity-relationship for lead optimization or by revealing on- and off-target sites to guide further cell-type specific in vitro chemical screening and optimization.

摘要 阿片类药物在减轻疼痛方面非常有效，但它们上瘾和过量服药的可能性导致越来越多的人 公共卫生危机。研究人员试图开发新的阿片类化合物，这种化合物不太可能 被滥用且副作用较少，但这些努力一直很困难。内源性阿片系统有 多种受体和配体在身体不同部位和不同细胞类型中异质性表达。 人们已经做了大量的工作来描述阿片受体(OR)和配体之间的关系。 然而，配体-受体结合的特异性通常取决于预先确定的相对亲和力。 目标。总体而言，在体内大脑空间和细胞的异质性掩盖了阿片类药物的作用，使它们 仅凭受体亲和力很难预测。 目前，单细胞和空间转录学正在改变我们对大脑结构的理解。 然而，在我们如何衡量阿片类药物的行动并将其与空间分辨的 大脑的细胞图谱。利用新兴的捕获和反向活动标记(IAM)技术，我们提出了 通过空间和单细胞分辨率对整个小鼠大脑的阿片类药物作用进行多模式分析。vbl.使用 三种药理上不同的阿片类药物，我们的目标是绘制这些药物的神经元活动和细胞结合图 以一种不偏不倚的方式移植到整个小鼠脑中，并将它们与从单个细胞识别的细胞类型进行注册 转录学。此外，我们将测试一种药物在不同ORs之间的亲和力是否决定其在体内的 细胞和神经整体的结合。该项目不仅将提供一种电路级机制，将 分子药理学对全脑阿片类药物的作用，也为评估和开发铺平了路线图 新的阿片类药物，例如通过将区域和细胞类型偏好纳入到结构-活性-关系中 引导优化或通过揭示靶点上和靶点外的位置来指导进一步的细胞类型特异性体外化学 筛选和优化。