权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Validation of FGF14 as a New Molecular Target of GSK3

验证 FGF14 作为 GSK3 的新分子靶点

基本信息

批准号：
9061830
负责人：
Fernanda Laezza
金额：
$ 38.25万
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：
2012
资助国家：
美国
起止时间：
2012-08-01 至 2018-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9061830
关键词：
Action Potentials Addictive Behavior Binding Biochemical Biochemistry Biological Assay Biology Bioluminescence Brain Cell Culture System Cells Chronic Co-Immunoprecipitations Complex Confocal Microscopy Data Development Disease Drug Targeting Electrophysiology (science)Enzymes Functional disorder Glycogen Synthase Kinase 3 Hippocampus (Brain)Image Intervention Life Link Maintenance Mass Spectrum Analysis Mental Depression Mental disorders Modeling Molecular Molecular Biology Molecular Target Mood Disorders Neuronal Dysfunction Neuronal Plasticity Neurons Outcome Study Pathway interactions Peptides Pharmaceutical Preparations Pharmacotherapy Phenotype Phosphorylation Proto-Oncogene Proteins c-akt Repressor Proteins Rodent Model Role Schizophrenia Site Site-Directed Mutagenesis Slice Synapses Synaptic Transmission Technology Testing Therapeutic Intervention Time Validation base cellular imaging cortico-limbic circuits fibroblast growth factor-14 genetic approach in vivo innovation insight multidisciplinary mutant neuroadaptation neuronal circuitry neuronal excitability novel patch clamp prevent psychiatric symptom screening success tool

项目摘要

DESCRIPTION (provided by applicant): Psychiatric diseases are chronic, devastating disorders thought to arise from maladaptive brain plasticity, and potent and safe pharmacotherapies are in great need. Identifying the mechanistic links that might sustain these aberrant neuroadaptations will advance our understanding of the biology of mental disorders, potentially providing new platforms for medication development. Using an innovative bioluminescence-based molecular screening approach combined with biochemical, electrophysiological, and imaging assays, we provide breakthrough results showing a link between glycogen synthase kinase 3 (GSK3), a critical enzyme found dysfunctional in mood disorders, depression and schizophrenia, and neuronal excitability, which we propose as a potential mechanism underlying dysfunction of neuronal circuitries associated with psychiatric disorders and certain addictive behaviors. Building on previous discoveries demonstrating that fibroblast growth factor 14 (FGF14) is a functionally relevant component of the Nav channelosome that controls neuronal excitability, we present exciting new data showing that the FGF14:Nav channel complex formation is bi-directionally controlled by GSK3 and by the GSK3 constitutive repressor, protein kinase B (Akt), and that GSK3 directly phosphorylates FGF14. Pharmacological inhibition of Akt and GSK increases and prevents, respectively, the FGF14:Nav channel complex formation, whereas inhibition of GSK3 occludes the effect of Akt inhibition. In hippocampal neurons, GSK3 inhibition disperses the FGF14:Nav channel complex from the axonal initial segment (AIS), the site of action potential initiation, impairs intrinsic fring and reduces excitatory synaptic transmission, whereas inhibition of Akt leads to opposite phenotypes. Furthermore, we show that Fpep1, a small interfering peptide modeled upon the FGF14:Nav channel interface, prevents the FGF14:Nav channel complex assembly, providing a tool for minimizing the effect of GSK3 on neuronal excitability in vivo. In this proposal we will employ a combination of bioluminescence-based technology, mass spectrometry, phosphorylation assays, confocal imaging and electrophysiology to determine the molecular mechanism by which GSK3 controls the FGF14:Nav channel complex formation (Aim 1) and promotes targeting of the FGF14:Nav channel complex in neurons (Aim 2) and to evaluate whether GSK3 exerts an effect on excitability and neuroplasticity in cortico-limbic circuits through the FGF14:Nav channel complex that could be reversed by pharmacological or genetic approaches targeting FGF14 (Aim 3). Positive outcomes of this study will provide new insights into the molecular mechanisms of GSK3 in the brain and offer an unprecedented opportunity for new medication development against GSK3-linked psychiatric disorders.

描述（由申请人提供）：精神疾病是一种慢性、破坏性的疾病，被认为是由大脑可塑性适应不良引起的，非常需要有效和安全的药物治疗。确定可能维持这些异常神经适应的机制联系将促进我们对精神障碍生物学的理解，可能为药物开发提供新的平台。使用创新的基于生物发光的分子筛选方法，结合生物化学，电生理和成像分析，我们提供了突破性的结果，显示糖原合成酶激酶3（GSK 3），一种在情绪障碍，抑郁症和精神分裂症中发现功能障碍的关键酶，与神经元兴奋性之间的联系，我们认为这是与精神疾病和某些成瘾行为相关的神经回路功能障碍的潜在机制。基于先前的发现表明成纤维细胞生长因子14（FGF 14）是控制神经元兴奋性的Nav通道体的功能相关组分，我们提出了令人兴奋的新数据，其显示FGF 14：Nav通道复合物的形成由GSK 3和GSK 3组成性阻遏物蛋白激酶B（Akt）双向控制，并且GSK 3直接磷酸化FGF 14。Akt和GSK的药理学抑制分别增加和防止FGF 14：Nav通道复合物形成，而GSK 3的抑制封闭Akt抑制的作用。在海马神经元中，GSK3抑制将FGF 14：Nav通道复合物从轴突起始段（AIS）（动作电位起始位点）分散，损害内在的fring并减少兴奋性突触传递，而Akt的抑制导致相反的表型。此外，我们表明，Fpep1，一个小的干扰肽模拟FGF 14：Nav通道接口，防止FGF 14：Nav通道复合物组装，提供了一个工具，最大限度地减少GSK 3对神经元兴奋性的影响在体内。在本提案中，我们将采用基于生物发光的技术、质谱法、磷酸化测定、共聚焦成像和电生理学的组合来确定GSK 3控制FGF 14：Nav通道复合物形成（目的1）和促进神经元中FGF 14：Nav通道复合物靶向（目的2）的分子机制，并评估GSK 3是否通过FGF 14：Nav通道复合物对皮质边缘回路的兴奋性和神经可塑性产生影响，该影响可通过靶向FGF 14的药理学或遗传学方法逆转（目的3）。这项研究的积极成果将为GSK3在大脑中的分子机制提供新的见解，并为针对GSK3相关精神疾病的新药开发提供前所未有的机会。