Novel tool development for quantitative PharmacoSTORM super-resolution imaging of the nanoscale distribution of D3 dopamine receptors

用于 D3 多巴胺受体纳米级分布定量 PharmacoSTORM 超分辨率成像的新工具开发

• 批准号: 10670438
• 负责人: Istvan Katona
• 金额: $ 18.55万
• 依托单位: TRUSTEES OF INDIANA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10670438
• 关键词: Acute; Adult; Adverse effects; Affect; Agonist; American; Anatomy; Animal Model; Antibodies; Antidepressive Agents; Antipsychotic Agents; Anxiety; Axon; Binding; Binding Sites; Brain; Brain Diseases; Brain imaging; Brain region; COVID-19 pandemic; Cell membrane; Cells; Chemicals; Clinical; Communication; Confocal Microscopy; Data Analyses; Development; Diazomethane; Dissociation; Dopamine Receptor; Drug usage; Electrophysiology (science); Emotional; Exhibits; FDA approved; G-Protein-Coupled Receptors; Goals; Health; Hippocampus; Human; Image; Impairment; Incubated; Islands of Calleja; Knockout Mice; Lead; Legal; Ligand Binding; Ligands; Lighting; Measurement; Mediating; Medicine; Mental Depression; Mental Health; Mental disorders; Methodology; Molecular; Mus; National Institute of Drug Abuse; Nature; Neurobiology; Neurons; Neurosciences; Nucleus Accumbens; Pathologic; Pathologic Processes; Patients; Pharmaceutical Preparations; Pharmacology; Photoaffinity Labels; Physiological; Plasma Cells; Prefrontal Cortex; Presynaptic Terminals; Prevalence; Process; Property; Protocols documentation; Relapse; Rodent; Schizophrenia; Signal Transduction; Site; Slice; Substance Use Disorder; Surveys; System; Therapeutic; Therapeutic Effect; Time; Tissues; Ultraviolet Rays; Ventral Tegmental Area; Visualization; Withdrawal Symptom; Work; addiction; analog; antagonist; cell type; clinically significant; comorbidity; cyanine dye 5; data acquisition; density; design; dopamine D3 receptor; dopaminergic neuron; drug seeking behavior; granule cell; hippocampal pyramidal neuron; imaging approach; imaging modality; in vivo; innovation; nanoscale; neurobiological mechanism; neuropsychopharmacology; novel; opioid epidemic; patch clamp; pharmacologic; prevent; receptor; receptor-mediated signaling; small molecule; social; substance use; substance use treatment; superresolution imaging; tool; tool development; trend; ultra high resolution

Abstract Substance use disorders affect ~20 million American adults according to the National Survey on Drug Use and Health. About 8.5 million patients also suffer from a comorbid mental health disorder such as depression, anxiety, and schizophrenia. Moreover, the prevalence of illicit substance use, the misuse of legal substances, and other mental health illnesses are all rapidly increasing in association with the Covid-19 pandemic. Thus, mechanistic understanding of the pathological neurobiological processes underlying mental health and substance use disorders, and their co-morbidity is imperative for the development of more efficient therapies to reduce physical, emotional, and social harm. It is widely acknowledged that dopaminergic signaling is highly impaired in these brain disorders. While the dopaminergic system has been intensely investigated in the last decades, it’s tremendous molecular and cellular complexity represents a major challenge for the detailed understanding of why certain treatments affecting the dopaminergic system are therapeutically beneficial or cause adverse effects. For example, pharmacological tools such as partial agonists, positive and negative allosteric modulators, and antagonists of the D3 dopamine receptor have been proposed by the National Institute of Drug Abuse as highly promising therapeutic tools for the treatment of substance use disorders. However, our understanding of the adaptive and maladaptive plasticity mechanisms controlled by D3 dopamine receptors and how this G protein- coupled receptor is involved in substance use disorders and other psychiatric diseases have remained very limited. This is an important, because emerging therapeutic trends, for example the expanding use of the FDA- approved D3 receptor-preferring drug cariprazine (VraylarTM) in the treatment of depression and schizophrenia and its putative usefulness in the treatment of comorbid substance use disorders illuminates the clinical significance of D3 receptors. However, the generally very low copy numbers of D3 receptors in neurons and the current lack of sensitive and specific antibodies to visualize their distribution and reorganization in association with pathological processes renders studies aiming to better understand the function of D3 dopamine receptors very difficult. Therefore, the proposed work will deliver novel fluorescent tools that will enable cell-type- and subcellular compartment-specific quantification of D3 receptor distribution at the nanoscale level in intact brain circuits affected in substance use disorders. In preliminary studies, we developed the PharmacoSTORM super- resolution nanoscale pharmacology approach and applied fluorescent cariprazine to discover high cariprazine binding density in the Islands of Calleja. In the proposed R21 CEBRA, we will complete two specific aims: Aim 1. Develop photoaffinity-based fluorescent chemical probes for high-yield PharmacoSTORM super- resolution imaging of D3 dopamine receptors. Aim 2. Establish photoaffinity labeling for PharmacoSTORM imaging of low copy numbers of D3 dopamine receptors on specific neuronal types implicated in addiction.

抽象的 根据全国药物使用和滥用调查，药物滥用障碍影响约 2000 万美国成年人 健康 大约 850 万名患者还患有抑郁、焦虑等共病精神疾病。 此外，非法药物的使用、合法药物的滥用以及其他疾病的流行。 与 Covid-19 大流行相关的心理健康疾病都在迅速增加，因此，机械性的。 了解心理健康和物质使用背后的病理神经生物学过程 疾病及其共病对于开发更有效的疗法来减少身体、 人们普遍认为，这些疾病中的多巴胺能信号传导受到严重损害。 虽然过去几十年来对多巴胺能系统进行了深入研究，但它仍然存在。 巨大的分子和细胞复杂性对详细理解 为什么影响多巴胺能系统的某些治疗方法有益或引起不良反应。 例如，药理学工具，例如部分激动剂、正向和负向变构调节剂，以及 美国国家药物滥用研究所提出，D3 多巴胺受体拮抗剂具有高度 然而，我们对药物使用障碍的理解还不够深入。 D3 多巴胺受体控制的适应性和适应不良可塑性机制以及这种 G 蛋白如何- 偶联受体与物质使用障碍和其他精神疾病有关仍然非常严重 这是一个重要的因素，因为新兴的治疗趋势，例如 FDA 的扩大使用。 批准 D3 受体偏好药物卡利拉嗪 (VraylarTM) 用于治疗抑郁症和精神分裂症 及其在治疗共病物质使用障碍中的假定用途阐明了临床 然而，神经元中 D3 受体的拷贝数通常非常低。 目前缺乏敏感和特异的抗体来可视化它们的分布和关联重组 病理过程使得研究旨在更好地了解 D3 多巴胺受体的功能 因此，拟议的工作将提供新颖的荧光工具，使细胞类型和细胞类型成为可能。 完整大脑中纳米级 D3 受体分布的亚细胞区室特异性定量 在初步研究中，我们开发了 PharmacoSTORM super- 分辨率纳米级药理学方法并应用荧光卡利拉嗪发现高卡利拉嗪 卡列哈群岛的结合密度 在拟议的 R21 CEBRA 中，我们将完成两个具体目标： 目标 1. 开发基于光亲和力的荧光化学探针，用于高产量 PharmacoSTORM super- D3 多巴胺受体的分辨率成像。 目标 2. 为低拷贝数 D3 多巴胺的 PharmacoSTORM 成像建立光亲和标记 与成瘾有关的特定神经元类型上的受体。