Discovery of Chemical Probes for Psychiatric Disorders and Addiction

发现精神疾病和成瘾的化学探针

• 批准号: 9353468
• 负责人: Fernanda Laezza
• 金额: $ 54.89万
• 依托单位: UNIVERSITY OF TEXAS MED BR GALVESTON
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-15 至 2019-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9353468
• 关键词: Acute; Addictive Behavior; Affective; Alpha Cell; Animal Model; Animals; Anxiety; Area; Behavior; Biological; Biological Assay; Biology; Bioluminescence; Bipolar Disorder; Brain; Brain Diseases; Cell surface; Cells; Chemicals; Circular Dichroism; Classification; Clinical; Cognition Disorders; Cognitive; Communities; Complement; Complex; Computer Simulation; Data; Development; Disease; Drug Design; Electrophysiology (science); Environment; Foundations; Functional disorder; Future; Genetic; Glycogen Synthase Kinase 3; Goals; In Vitro; Ion Channel; Lead; Link; Luciferases; Maps; Measures; Membrane; Mental Depression; Mental disorders; Modality; Molecular; Molecular Probes; Mood Disorders; Mus; Neurons; Neuropharmacology; Nucleus Accumbens; Pathway interactions; Peptides; Permeability; Pharmaceutical Chemistry; Pharmaceutical Preparations; Pharmacology; Phenotype; Physiological; Process; Property; Protein Kinase; Proteins; Psychiatry; Research; Research Domain Criteria; Research Priority; Schizophrenia; Signal Transduction; Slice; Structure-Activity Relationship; Surface Plasmon Resonance; Tail; Testing; Therapeutic; Toxic effect; Transgenic Animals; Transgenic Mice; Validation; addiction; analog; base; brain circuitry; chemical synthesis; cognitive performance; design; drug discovery; fibroblast growth factor-14; flexibility; high throughput screening; improved; in vivo; innovation; interest; neural circuit; neuronal excitability; novel; overexpression; patch clamp; protein protein interaction; psychologic; reconstitution; response; screening; small molecule; stable cell line; therapeutic development; tool; trafficking; voltage

ABSTRACT Psychiatric diseases and addictive behaviors are complex brain disorders associated with maladaptive plasticity of the brain circuitry. The lack of adequate platforms to rapidly screen against novel targets in physiological environments has significantly hampered probe discovery initiatives that could inform the circuitry alterations leading to mental disorders and addiction and enable the design of future therapeutics. Protein- protein interactions (PPI) within ion channel complexes fine-tune neuronal excitability and are emerging as links to the biology of psychiatric disorders. Their highly specific and flexible interfaces could make protein- channel interactions ideal targets for probe development. Such molecular probes would provide the neuropharmacology community with optimal research tools to parse out brain disease complexities and ultimately enable future drug design. We have identified the PPI between the voltage-gated Na+ (Nav) Nav1.6 channel and its accessory regulator protein, fibroblast growth factor 14 (FGF14) as a novel, functionally relevant regulator of neuronal excitability in brain areas such as the cortico-mesolimbic circuit, which is associated with disorders of the affective and cognitive domains. Through a successful bioluminescence-based high-throughput screen (HTS) and subsequent in vivo studies in the nucleus accumbens (NAc), we discovered that the FGF14:Nav1.6 channel complex is part of the glycogen synthase kinase 3 (GSK3) pathway, a signaling cascade found aberrant in bipolar disorder, depression, anxiety and addiction. To explore the druggability of the FGF14:Nav1.6 complex, we employed a minimal functional domain (MFD) approach to design a peptide-derivative mapped to the PPI interface and showed it has in vitro-to-ex vivo activity in the NAc circuit. These discoveries have prompted us to develop a new pipeline to identify chemical probes against the FGF14:Nav1.6 complex to interrogate its function in the cortico-mesolimbic circuit. To advance a probe discovery campaign against this new target, we have designed an integrated multi-modal screening platform, based on the latest MFD principles of pharmacology that includes a newly designed and validated bioluminescence primary screening assay to reconstitute the FGF14:Nav1.6 C-tail complex in cells. This pipeline also includes the necessary counter, toxicity, and cell-free orthogonal assays (Aim 1), automated patch-clamp electrophysiology as a functional screen in combination with structure-activity relationship efforts and in silico analysis (Aim 2), and ex vivo validation of selected probes in the NAc circuitry (Aim 3). The proposed pipeline introduces a new, rapid and integrated platform that will accelerate the discovery of novel chemical probes for neuronal excitability, providing the foundation for pre-therapeutic development of a new class of PPI-based leads for a broad spectrum of psychiatric disorders.

抽象的 精神疾病和成瘾行为是与适应不良有关的复杂脑部疾病 脑电路的可塑性。缺乏足够的平台来快速筛选针对新的目标 生理环境严重阻碍了可以告知电路的探针发现计划 改变导致精神障碍和成瘾的改变，并使未来的治疗剂的设计。蛋白质- 离子通道复合物中的蛋白质相互作用（PPI）可微调神经元兴奋性，并随着 链接到精神疾病的生物学。它们的高度特异性和柔性界面可能会使蛋白质 - 通道相互作用理想的探针开发目标。这样的分子探针将提供 神经药理学界具有最佳的研究工具，可以解析脑部疾病复杂性和 最终使未来的药物设计。 我们已经确定了电压门控的NA+（NAV）NAV1.6通道及其附件调节器之间的PPI 蛋白质，成纤维细胞生长因子14（FGF14）作为一种新型，功能相关的神经元兴奋性调节剂 大脑区域（例如Cortico-Mesolimbic电路）与情感和情感疾病有关 认知领域。通过成功基于生物发光的高通量屏幕（HTS）和 随后在伏隔核（NAC）的体内研究中，我们发现FGF14：NAV1.6通道 复合物是糖原合酶激酶3（GSK3）途径的一部分，这是一种信号级联，发现在 躁郁症，抑郁，焦虑和成瘾。探索FGF14：NAV1.6复合物的吸毒性， 我们采用了最小功能域（MFD）方法来设计映射到PPI的肽衍生物 界面并显示其在NAC电路中具有体外到EX的体内活性。这些发现促使我们 开发一条新管道来识别针对FGF14的化学探针：NAV1.6复合物以询问其 在Cortico-Mesolimbic电路中的功能。为了推进针对这个新目标的调查发现运动，我们 已经根据最新的MFD原理设计了一个集成的多模式筛选平台 药理学包括新设计和经过验证的生物发光初级筛选测定法 重建FGF14：细胞中的NAV1.6 C-Tail复合物。该管道还包括必要的计数器 毒性和无细胞的正交分析（AIM 1），自动贴片钳电生理学作为功能 屏幕与结构活性关系工作和计算机分析（AIM 2）结合使用（AIM 2）和离体 NAC电路中选定的探针的验证（AIM 3）。拟议的管道引入了一个新的，快速且 集成平台将加速发现新颖的化学探针以实现神经元兴奋性， 为一类新的基于PPI的潜在客户提供了治疗前开发的基础 精神疾病的范围。