Role of protein synthesis in Alzheimers disease-associated impairments of synaptic plasticity and memory

蛋白质合成在阿尔茨海默病相关的突触可塑性和记忆损伤中的作用

基本信息

批准号：
10180826
负责人：
Tao Ma
金额：
$ 46.43万
依托单位：
WAKE FOREST UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-08-01 至 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10180826
关键词：
AD transgenic mice APP-PS1 Alzheimer&apos s Disease Alzheimer&apos s disease brain Alzheimer&apos s disease model Amino Acids Amyloid beta-Protein Biochemical Biological Assay Brain Cognition Disorders Cognitive Cognitive deficits Data Defect Dementia Deterioration Development Disease Electrophysiology (science)Etiology Event Failure Foundations Frontotemporal Dementia Functional disorder Future Genetic Genetic Translation Goals Hippocampus (Brain)Human Imaging Techniques Impairment Knock-out Knowledge Learning Location Long-Term Potentiation Mass Spectrum Analysis Mediating Memory Memory impairment Messenger RNA Methods Modeling Molecular Mus Neurodegenerative Disorders Neurosciences Pathogenesis Peptide Elongation Factor 2 Pharmacology Phase Phosphorylation Phosphotransferases Prion Diseases Protein Biosynthesis Proteins Proteomics Reporting Repression Research Ribosomes Role Sampling Signal Transduction Site Slice Structure Surface Synapses Synaptic plasticity Syndrome Testing Therapeutic Translations Work base behavior test confocal imaging design diagnostic biomarker effective therapy experimental study genetic approach genetic testing improved inhibitor/antagonist knock-down morris water maze mouse genetics mouse model novel novel diagnostics peptidyl-tRNA prevent restoration small molecule inhibitor synaptic failure synaptic function therapeutic target water maze

项目摘要

The basic cellular/molecular signaling mechanisms underlying Alzheimer’s disease (AD) pathophysiology are not well understood; this gap in knowledge is hampering our ability to find any effective therapies. Accumulating evidence indicates impaired synaptic function as a key event in AD pathogenesis. However, the molecular mechanisms underlying AD-associated synaptic dysfunction/failure remain elusive. We recently reported hyperphosphorylation of mRNA translational factor eukaryotic elongation factor 2 (eEF2) in AD brains. Phosphorylation of eEF2 by its (only known) kinase eEF2K results in repression of de novo protein synthesis, which is essential for long-lasting forms of synaptic plasticity and memory. Driven by the preliminary data, the central hypothesis to be tested in this application is that restoration of the capacity for de novo protein synthesis, via inhibition of eEF2K and thus eEF2 phosphorylation, will alleviate AD-associated synaptic failure and memory impairments. Three specific aims have been designed to test this hypothesis. Aim 1 seeks to determine whether restoration of normal eEF2 phosphorylation, via suppressing eEF2K activity, can rescue AD-associated impairments in hippocampal long-term synaptic plasticity. Aim 2 is to determine whether inhibition of eEF2K activity improves learning and memory deficits in AD mouse model. Aim 3 is to determine whether AD-associated impairments of de novo protein synthesis can be mitigated by inhibiting eEF2 kinase activity. The project proposes in-depth analyses using multiple state-of-art methods in neuroscience, including synaptic electrophysiology, confocal imaging, mouse genetics, and behavioral tests. We will also employ two new types of non-radioactive methods to assess de novo protein synthesis in brain slices: surface sensing of translation (SUnSET) and bioorthogonal noncanonical amino acid tagging (BONCAT). These novel methods will be combined with mass spectrometry/proteomics approach to reveal identities of proteins in AD brains whose synthesis is dysregulated because of abnormal eEF2K/eEF2 signaling. Findings from this project will contribute important data regarding the cellular/molecular signaling mechanisms underlying AD pathogenesis. Future studies will build on the results from this project and our other research findings on AD-related protein synthesis dysregulation to inform eventual development of novel diagnostic markers and better therapeutic strategies for AD-related cognitive syndromes, for which no effective treatments exist.

阿尔茨海默氏病（AD）病理生理的基本细胞/分子信号传导机制是不太了解；知识差距正在阻碍我们找到任何有效疗法的能力。积累的证据表明突触功能受损是AD发病机理的关键事件。但是，与AD相关的突触功能障碍/失效的分子机制仍然难以捉摸。我们最近报道了AD大脑中mRNA转移因子真核伸长因子2（EEF2）的热磷酸化。 EEF2通过其（仅已知的）激酶EEF2K磷酸化导致从头蛋白质合成的表达，这对于突触可塑性和记忆的持久形式至关重要。由初步数据驱动在此应用中要测试的中心假设是恢复从头蛋白的能力通过抑制EEF2K并因此，合成将减轻与AD相关的突触衰竭和记忆障碍。已经设计了三个特定目标来检验这一假设。目标1寻求确定是否通过抑制EEF2K活性来恢复正常的EEF2磷酸化是否可以营救海马长期突触可塑性中与广告相关的损伤。目标2是确定是否 EEF2K活性的抑制可改善学习和记忆在AD鼠标模型中定义。目标3是确定通过抑制EEF2激酶，可以减轻从头蛋白合成的AD相关损伤活动。该项目建议使用神经科学中的多种最新方法进行深入分析，包括突触电生理学，共聚焦成像，小鼠遗传学和行为测试。我们还将雇用两个新型的非放射性活性方法评估脑切片中从头蛋白质合成的新类型：表面敏感性翻译（日落）和生物正交的非规范氨基酸标记（Boncat）。这些新颖的方法将与质谱/蛋白质组学方法结合使用，以揭示AD大脑中蛋白质的身份由于EEF2K/EEF2信号传导异常，其合成失调。这个项目的发现将关于AD发病机理的细胞/分子信号传导机制的重要数据。未来的研究将基于该项目的结果以及我们关于广告相关蛋白的其他研究结果合成失调，以告知新型诊断标记和更好治疗的最终发展与广告相关的认知综合征的策略，不存在有效治疗。