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: 10510936
• 负责人: Istvan Katona
• 金额: $ 22.71万
• 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/10510936
• 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 (Brain); Human; Image; Impairment; Islands of Calleja; Knockout Mice; Lead; Legal; Ligand Binding; Ligands; Lighting; Measurement; Mediating; Medicine; Mental Depression; Mental Health; Mental disorders; Methodology; Methods; 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; Resolution; 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; base; 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; prevent; receptor; receptor-mediated signaling; small molecule; social; substance use; substance use treatment; tool; tool development; trend

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 根据全国药物使用调查和 健康。大约850万患者也患有合并症的精神健康障碍，例如抑郁，焦虑， 和精神分裂症。此外，非法物质使用的普遍性，法律物质的滥用和其他 与19日大流行有关的心理健康疾病都在迅速增加。那，机械 理解心理健康和药物使用的病理神经生物学过程 疾病及其合并症对于开发更有效的疗法以减少身体的疗法至关重要 情感和社会伤害。人们普遍认为，多巴胺能信号在这些信号中受到了极大的损害 脑疾病。在过去的几十年中，多巴胺能系统已经积极研究 巨大的分子和细胞复杂性代表了对详细理解的主要挑战 为什么某些影响多巴胺能系统的治疗方法是热有益或引起不利影响。 例如，药物工具，例如部分激动剂，正和负变构调节剂，以及 美国国家药物滥用研究所提出了D3多巴胺受体的拮抗剂 有希望的治疗药物使用障碍的治疗工具。但是，我们对 由D3多巴胺受体控制的自适应和不良适应性可塑性机制以及该G蛋白如何 耦合受体参与药物使用障碍，其他精神病疾病仍然非常 有限的。这是一个重要的，因为新兴疗法的趋势，例如FDA-的扩展使用 批准的D3受体挑选药物甲生花（Vraylartm）用于治疗抑郁和精神分裂症 它在合并症使用障碍的治疗中的假定有用性阐明了临床 D3受体的重要性。但是，神经元中的D3受体的通常非常低的拷贝数和 当前缺乏敏感和特定的抗体可视化其分布和重组 通过病理过程，旨在更好地了解D3多巴胺受体的功能的研究 非常困难。因此，拟议的工作将提供新型的荧光工具，以实现细胞类型和 D3受体分布在完整脑中D3受体分布的亚细胞区室特异性定量 在药物使用障碍中受影响的电路。在初步研究中，我们开发了Pharmacostorm超级 分辨率纳米级药理学方法和应用的荧光宫嗪可发现高甲氧氨基 Calleja岛的结合密度。在拟议的R21 CEBRA中，我们将完成两个具体的目标： 目标1。开发基于光附近的荧光化学问题 D3多巴胺受体的分辨率成像。 目标2。建立低拷贝数D3多巴胺的药物遗传学成像的光性标签 成瘾中实施的特定神经元类型的受体。