Tau-Fyn Interaction and Alzheimer's Disease

Tau-Fyn 相互作用与阿尔茨海默病

• 批准号: 10292907
• 负责人: Jonathan R Roth
• 金额: $ 1.62万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2021-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10292907
• 关键词: 3-Dimensional; AMPA Receptors; Ablation; Affect; Affinity; Age; Alzheimer' s Disease; Alzheimer' s disease model; Alzheimer' s disease patient; Alzheimer' s disease therapeutic; Amyloid beta-Protein; Automobile Driving; Behavioral Assay; Binding; Biological Assay; Biological Models; Cognitive; Cognitive deficits; Dementia; Dendrites; Dendritic Spines; Disease Progression; Electroencephalography; Epilepsy; Functional disorder; Genetic; Glutamates; Goals; Hippocampus (Brain); Impaired cognition; In Situ; In Vitro; Knock-out; Knockout Mice; Lead; Learning; Ligation; MAPT gene; Measures; Mediating; Mediator of activation protein; Membrane; Modeling; Molecular; Morphology; Mus; Mutate; Mutation; N-Methyl-D-Aspartate Receptors; N-Methylaspartate; Nerve Degeneration; Neurites; Neurofibrillary Tangles; Neuronal Dysfunction; Neurons; PXXP Motif; Pathogenesis; Peptides; Phenotype; Phosphorylation; Phosphotransferases; Population; Predisposition; Proline-Rich Domain; Proteins; Proto-Oncogene Proteins c-fyn; Public Health; Role; SH3 Domains; Scientist; Seizures; Signal Transduction; Structure; Synapses; System; Techniques; Testing; Therapeutic; Vertebral column; Work; abeta toxicity; base; behavior test; beta amyloid pathology; career; cognitive testing; density; excitotoxicity; extracellular; familial Alzheimer disease; gabazine; hyperphosphorylated tau; immunocytochemistry; in vivo; inhibitor/antagonist; insight; member; mouse model; multi-electrode arrays; novel strategies; novel therapeutic intervention; osmotic minipump; overexpression; postsynaptic; prevent; relating to nervous system; skills; tau Proteins; tau interaction; therapeutic target; therapy development; trafficking

Project Summary/Abstract Alzheimer’s disease (AD) is the leading cause of dementia worldwide and its impact will increase exponentially as the population ages. New therapeutic approaches are desperately needed to treat AD. The microtubule- associated protein Tau is has been heavily studies because it aggregates into neurofibrillary tangles within neurons, one of the hallmarks of AD. Genetic knockout of Tau is protective in several models of AD, highlighting its potential as a therapeutic target for AD. Interestingly, Tau reduction also prevents network hyperexcitability, which occurs early in AD and may contribute to neurodegeneration, in these models. Similarly, in a primary neuron culture system, Tau reduction prevents amyloid-β (Aβ) toxicity and glutamate or NMDA-induced excitotoxicity. In short, Tau reduction is protective in a variety of systems, but the mechanism by which it does so is currently unknown. Tau’s central proline-rich region, which is hyperphosphorylated in AD, has several PxxP motifs that mediate binding with SH3 domain–containing proteins including the nonreceptor tyrosine kinase Fyn. Fyn is also an important mediator of network hyperexcitability, as it phosphorylates AMPA and NMDA receptors to strengthen their signaling and regulates dendritic spine dynamics. Exogenous Aβ activates Fyn at the postsynaptic density, leading to NMDAR phosphorylation and excitotoxicity in neurons. Diverse evidence indicates that Tau’s interaction with Fyn could be a critical mediator of Aβ toxicity. Genetic knockout of either Tau or Fyn prevents Aβ toxicity in primary neurons, hyperphosphorylated Tau has a higher affinity for Fyn, and Tau mediates trafficking of Fyn to dendrites in vivo, leading to Aβ-induced cognitive deficits and network hyperexcitability. However, the idea that the Tau-Fyn interaction is a critical mediator of Aβ toxicity has not been directly tested, which is the goal of this project. My overarching hypothesis is that the Tau-Fyn interaction is a critical mediator of Aβ-induced structural and functional abnormalities. I will test this hypothesis using a peptide inhibitor of the Tau-Fyn interaction, Tau-PxxP5/6, developed by previous members of the Roberson lab. I developed a proximity ligation-based assay to confirm that Tau-PxxP5/6 inhibits endogenous Tau-Fyn interaction in situ and found that it prevents Aβ-induced neurite degeneration and membrane trafficking dysfunction. Here, I will determine if Tau-PxxP5/6 prevents Aβ-induced deficits in dendritic spine morphology using 3D morphometric analysis and Aβ- and gabazine-induced network hyperexcitability using multi-electrode arrays. I will also determine if the Fyn-binding region of Tau is the critical region of Tau that mediates Aβ toxicity by transducing Tau knockout neurons with different mutated Tau constructs to prevent Fyn binding. In addition to in vitro studies, I will determine if Tau-PxxP5/6 prevents cognitive deficits, epileptiform activity and seizure susceptibility in vivo using the hAPPJ20 mouse model of AD. The proposed work will provide insights into the molecular mechanisms of Aβ toxicity and could provide a promising therapeutic strategy to treat AD.

项目摘要/摘要 阿尔茨海默病(AD)是世界范围内导致痴呆的主要原因，其影响将呈指数级增长 随着人口老龄化。迫切需要新的治疗方法来治疗AD。微管- 相关蛋白Tau is已经得到了大量的研究，因为它聚集在 神经元，阿尔茨海默病的标志之一。Tau基因敲除在几种AD模型中具有保护性，强调 它有可能成为AD的治疗靶点。有趣的是，降低Tau还可以防止网络过度兴奋， 在这些模型中，这发生在AD的早期，并可能导致神经退行性变。类似地，在初级数据库中 神经元培养系统，Tau降低可防止淀粉样蛋白-β(A-β)毒性和谷氨酸或N-甲基-D-天冬氨酸诱导 兴奋性毒性。简而言之，牛磺酸还原在各种系统中都是有保护作用的，但它的机制 做到这一点目前尚不清楚。在阿尔茨海默病中过度磷酸化的tau的中央富含脯氨酸的区域 几个PxxP基序介导与含SH3结构域的蛋白结合，包括非受体 酪氨酸激酶Fyn。Fyn也是网络超兴奋性的重要介质，因为它使AMPA磷酸化 和NMDA受体，以加强其信号转导和调节树突棘的动力学。外源Aβ 在突触后密度激活Fyn，导致NMDAR的磷酸化和神经元的兴奋性毒性。 不同的证据表明，Tau与Fyn的相互作用可能是Aβ毒性的关键调节因子。遗传 敲除Tau或Fyn可防止原代神经元中的β毒性，过度磷酸化的Tau具有更高的 Fyn的亲和力和Tau介导Fyn在体内向树突的运输，导致β诱导的认知障碍 和网络的过度兴奋。然而，认为Tau-Fyn相互作用是β毒性的关键媒介的想法 还没有直接测试，这是这个项目的目标。我最重要的假设是Tau-Fyn 相互作用是Aβ诱导的结构和功能异常的关键介质。我要测试一下这个 使用Tau-Fyn相互作用的多肽抑制剂Tau-PxxP5/6的假说，由前成员开发 罗伯逊实验室。我建立了一种基于邻近连接的分析方法来证实Tau-PxxP5/6抑制内源性 Tau-Fyn相互作用的原位研究及发现它可阻止β诱导的轴突变性和膜转运 功能障碍。在这里，我将确定Tau-PxxP5/6是否能阻止Aβ诱导的树突棘形态缺陷 多电极三维形态计量学分析及β和Gabazine诱发的网络超兴奋性 数组。我还将确定Tau的FYN结合区是否是介导β毒性的Tau的关键区域 通过转导具有不同突变Tau结构的Tau基因敲除神经元来防止Fyn结合。此外 对于体外研究，我将确定Tau-PxxP5/6是否可以预防认知障碍、癫痫样活动和癫痫发作 体内易感性采用hAPPJ20小鼠AD模型。拟议的工作将提供对 Aβ毒性的分子机制，为AD的治疗提供了一种有前景的治疗策略。