Altered Copper Neuronal Transport and Alzheimer's Disease

铜神经元运输的改变与阿尔茨海默病

• 批准号: 10038512
• 负责人: Nicholas Sanchez
• 金额: $ 6.86万
• 依托单位: LOMA LINDA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-15 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10038512
• 关键词: Affect; Alleles; Alzheimer' s Disease; Alzheimer' s disease brain; Alzheimer' s disease model; Alzheimer' s disease pathology; Animal Model; Apolipoprotein E; Autopsy; Axon; Biological Assay; Brain; Brain region; Carrier Proteins; Cations; Cerebrospinal Fluid; Cognitive; Copper; Deposition; Diagnostic; Disease; Elements; Exhibits; Fluorescence; Fluorescent Probes; Goals; Hippocampus (Brain); Human; Human Pathology; Image; Imaging technology; Incidence; Investigation; Knock-in Mouse; Lipids; Measures; Memory; Metabolism; Mus; Neurons; Oxidative Stress; Pathogenesis; Pathologic Processes; Pathway interactions; Patients; Play; Positron-Emission Tomography; Production; Proteins; Reactive Oxygen Species; Research; Resources; Role; Specificity; Specimen; Synapses; Synaptosomes; Technology; Tissues; Transition Elements; United States National Institutes of Health; aging population; brain tissue; density; design; emission spectroscopy; innovation; insight; metal metabolism; mouse model; neuroblastoma cell; neuronal transport; neurotoxic; novel; synaptic pruning; tool

Project Summary I hypothesize that apoplioprotein E (APOE)-associated dysregulation of hippocampal copper (Cu) metabolism is a key step in Alzheimer’s disease (AD) pathogenesis. This project will approach this topic from two specific aims: 1) establish the relationship between copper (I) cation distribution and AD pathology, and 2) study the APOE4-alteration effect on hippocampal synaptic pruning and Cu deposition in the mouse hippocampus. This approach is innovative in its use of two newly developed technologies that will be used to investigate hippocampal Cu dysregulation, synaptic density and its relation to the toxic APOE4 allele. The CRISP-17 fluorescent probe is a state-of-the-art tool in histochemical Cu localization, with a specificity for monovalent Cu far beyond other existing probes. CRISP-17 will visualize Cu distribution in post- mortem human AD hippocampal brain tissue. Synaptosome fractions will be isolated and measured for Cu using atomic emissions spectroscopy. Cu transport proteins will be downregulated in SH-SY5Y neuroblastoma cells, with effects on Cu deposition being analyzed using the fluorescent probe and synaptosome fractions being isolated and measured for Cu levels. Synaptic density will be studied in APOE4 knock-in mice and compared with the neuroprotective APOE3 knock-in mice with PET scans targeting the SV2A synaptic protein. Mouse brain tissue will then be taken and histochemically analyzed for monovalent Cu using CRISP-17, verifying this relationship in the animal model. We expect to find similarly that APOE4 knock-in mice have a greatly reduced synaptic density when compared to the neuroprotective APOE3 knock-in mice, with the mouse hippocampus also exhibiting a decrease in Cu distribution. These aims are designed to further the goals of the NIH by investigating a possible pathway to AD, a disease whose pathogenesis not yet fully defined.

项目摘要 我推测载脂蛋白E(APOE)相关的海马铜调节失调 (铜)代谢是阿尔茨海默病(AD)发病机制中的关键步骤。这个项目将 从两个具体目标来探讨这一主题：1)建立铜与铜之间的关系(I) 阳离子分布与AD病理；2)研究载脂蛋白4改变对海马区的影响 突触修剪与小鼠海马铜沉积。这种方法在以下方面具有创新性 它使用了两项新开发的技术，这两项技术将用于研究海马铜 调节失调、突触密度及其与毒性APOE4等位基因的关系。CRISP-17 荧光探针是一种最先进的组织化学铜定位工具，具有特异性 单价铜远远超过其他现有的探针。CRISP-17将可视化铜在后部的分布 尸检人AD海马区脑组织。突触体组分将被分离并 使用原子发射光谱测量铜的含量。铜转运蛋白将是 SH-SY5Y神经母细胞瘤细胞中铜沉积的影响 用荧光探针和突触体组分分离和测定铜 级别。将研究APOE4基因敲入小鼠的突触密度，并与 针对SV2a突触蛋白的PET扫描的神经保护APOE3敲入小鼠。 然后取小鼠脑组织，用组织化学方法分析单价铜 CRISP-17，在动物模型中验证了这种关系。我们预计会发现类似的情况 与对照组相比，ApoE4基因敲入小鼠的突触密度显著降低 神经保护性APOE3基因敲入小鼠，小鼠海马区也表现出减少 在铜的分布上。这些目标旨在通过调查一项 阿尔茨海默病的可能途径，其发病机制尚未完全确定。