Discovery of Chemical Probes for Psychiatric Disorders and Addiction

发现精神疾病和成瘾的化学探针

基本信息

批准号：
9252858
负责人：
Fernanda Laezza
金额：
$ 51.83万
依托单位：
UNIVERSITY OF TEXAS MED BR GALVESTON
依托单位国家：
美国
项目类别：
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/9252858
关键词：
Acute Addictive Behavior Affective Animal Model Animals Anxiety Area Behavior Biological Biological Assay Biology Bioluminescence Bipolar Disorder Brain Brain Diseases Cell surface Cells Chemicals Circular Dichroism Classification Clinical Cognitive Communities 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 Protein Traffic Mental Depression Mental disorders Molecular Molecular Probes Mood Disorders Mus Neurons Neuropharmacology Nucleus Accumbens Pathway interactions Peptides Permeability Pharmaceutical Chemistry Pharmaceutical Preparations Pharmacology Physiological Process Property Proteins Psychiatry Research Research Priority SCN8A gene 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 voltage

项目摘要

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相关的复杂脑部疾病大脑回路的可塑性。缺乏足够的平台来快速筛选新目标生理环境极大地阻碍了可以为电路提供信息的探针发现计划导致精神障碍和成瘾的改变使未来疗法的设计成为可能。蛋白质- 离子通道复合物内的蛋白质相互作用（PPI）可微调神经元的兴奋性，并正在成为与精神疾病的生物学联系。它们高度特异性和灵活的界面可以使蛋白质通道相互作用是探针开发的理想目标。这种分子探针将提供神经药理学界拥有最佳研究工具来解析脑部疾病的复杂性和最终实现未来的药物设计。我们已经确定了电压门控 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) 作为一种新型的、功能相关的神经元兴奋性调节剂大脑区域，例如皮质-中脑边缘回路，与情感和情感障碍有关认知领域。通过成功的基于生物发光的高通量筛选（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，该途径是一种异常的信号级联反应双相情感障碍、抑郁、焦虑和成瘾。为了探索 FGF14:Nav1.6 复合物的成药性，我们采用最小功能域 (MFD) 方法来设计映射到 PPI 的肽衍生物界面并表明它在 NAc 回路中具有体外到离体活性。这些发现提示我们开发新的管道来识别针对 FGF14:Nav1.6 复合物的化学探针，以询问其在皮质-中脑边缘回路中发挥作用。为了推进针对这一新目标的探测器发现活动，我们基于最新的 MFD 原理，设计了一个集成的多模态筛选平台药理学，包括新设计和验证的生物发光初级筛选试验在细胞中重建 FGF14:Nav1.6 C 尾复合物。该管道还包括必要的计数器，毒性和无细胞正交测定（目标 1），自动膜片钳电生理学作为功能结合结构-活性关系工作和计算机分析（目标 2）和离体筛选验证 NAc 电路中选定的探针（目标 3）。拟议的管道引入了一种新的、快速的、集成平台将加速神经元兴奋性新型化学探针的发现，为广泛的基于 PPI 的新型先导药物的治疗前开发奠定基础一系列精神疾病。