Project 2: Co-pathogenic Interactions between ApoE Isoforms and Abeta in Neural Network Dysfunction of Alzheimer's Disease

项目 2：ApoE 同工型和 Abeta 在阿尔茨海默病神经网络功能障碍中的共致病相互作用

• 批准号: 10271127
• 负责人: Jorge J Palop
• 金额: $ 92.2万
• 依托单位: J. DAVID GLADSTONE INSTITUTES
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-15 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10271127
• 关键词: Aging; Alzheimer' s Disease; Alzheimer' s Disease Pathway; Alzheimer' s disease model; Amyloid beta-Protein; Amyloid beta-Protein Precursor; Apolipoprotein E; Astrocytes; Behavior; Behavioral; Brain; Candidate Disease Gene; Cells; Clinical Trials; Cognitive; Cognitive deficits; Collaborations; Complex; Dependence; Disease; Disease Progression; Electroencephalography; Electrophysiology (science); Etiology; Functional disorder; Gene Expression; Genotype; Histopathology; Human; Human Amyloid Precursor Protein; Impairment; Interneurons; Knock-in Mouse; MAPT gene; Machine Learning; Microglia; Molecular; Mus; Mutation; Neurons; Parvalbumins; Pathogenesis; Pathogenicity; Pathologic; Pathology; Pathway interactions; Physiological; Physiology; Protein Isoforms; Proteins; Somatostatin; Testing; Therapeutic; Transgenes; Wireless Technology; apolipoprotein E-4; cell type; circadian; cognitive function; in vivo; morris water maze; mouse model; network dysfunction; neural network; novel; optogenetics; overexpression; resilience; single-cell RNA sequencing; tau Proteins; transcriptome; transcriptomics

PROJECT 2 – SUMMARY Numerous lines of evidence suggest that human amyloid precursor protein/amyloid b (APP/Ab), apoE4, and the microtubule-associated protein tau contribute to Alzheimer’s disease (AD), but their pathogenic interactions are largely unknown. Reducing Ab accumulation in the brain was considered the most reasonable therapeutic strategy for AD, but multiple clinical trials of this approach have failed, suggesting that the pathophysiology of AD is much more complex than anticipated and that the pathogenic interactions of AD-relevant proteins need to be better understood. To decode the multifactorial etiology of AD at physiological levels of expression, we will study newly developed knock-in (KI) mouse models of late-onset (LOAD) and familial (FAD) AD without transgene overexpression and focus on the pathogenic interactions between Ab, apoE, and tau. To simulate LOAD, we will use KI mice that express humanized wildtype Ab without FAD mutations (ApphAβWT/hAβWT; referred to as Aβ mice), human apoE isoforms (APOEE2/E2, APOEE3/E3, and APOEE4/E4; E2, E3, and E4 mice), and human wildtype tau (MAPTWT/WT, TAUWT mice). To simulate FAD, we will use KI mice that express humanized wildtype Ab with the Swedish and Iberian FAD mutations (AppNL-F/NL-F; ↑Ab mice), human apoE isoforms (E3 and E4), and human wildtype tau (TAUWT). Thus, we propose to study the physiological and endogenous interactions of human Ab, apoE isoforms, and tau in vivo that contribute to AD-related abnormalities in neural network activity, cognitive functions, gene expression, and histopathology using state-of-the-art in vivo electrophysiological, optogenetics, and behavioral approaches. We will focus on mechanisms of altered neural network dysfunction, since they closely relate to brain and cognitive functions and are disrupted early in AD pathogenesis. In Aim 1, we will determine pathogenic interactions of Ab, apoE isoforms, and tau contributing to altered neural network activity (Aim 1a) and behavioral deficits (Aim 1b) in LOAD and FAD KI mice during disease progression using wireless long-term EEG/EMG recordings and standard and machine learning behavioral approaches. In Aim 2, we will determine pathogenic interactions of Ab and apoE4 contributing to cell and circuit function impairments in LOAD and FAD KI mice in vivo (Aim 2a, b) and if optogenetic activation of specific interneuron cell types reverses AD-related abnormalities (Aim 2c). We will use in vivo LFP and multi-unit recordings and optogenetic approaches in behaving mice to identify cell type and circuit-level mechanisms of network dysfunction. In Aim 3, we will determine pathogenic interactions of Ab, apoE isoforms, and tau contributing to AD-related pathology (Aim 3a) and scRNA-seq transcriptome changes (Aim 3b) in LOAD and FAD KI mice. We will perform AD-related pathological and scRNA-seq transcriptomics analyses in functionally characterized mice (in vivo physiology and behavior) from Aims 1 and 2 to identify pathological and transcriptome changes associated with cognitive vulnerability and resilience. We aim to identify human validated and behaviorally relevant candidate genes/pathways of AD pathogenesis and define cell-type-specific disease mechanisms.

项目2--摘要 大量证据表明，人类淀粉样前体蛋白/淀粉样蛋白b(APP/Ab)、apoE4和 微管相关蛋白tau参与阿尔茨海默病(AD)，但它们的致病作用是 很大程度上是未知的。减少抗体在大脑中的积聚被认为是最合理的治疗方法 治疗阿尔茨海默病的策略，但这种方法的多项临床试验都失败了，表明阿尔茨海默病的病理生理学 AD比预期的要复杂得多，AD相关蛋白的致病相互作用需要 更好地理解。为了在生理水平上破译阿尔茨海默病的多因素病因，我们将 研究新发展的晚发性(LOAD)和家族性(FAD)AD敲入(KI)小鼠模型 转基因过表达，并关注抗体、载脂蛋白E和tau之间的致病相互作用。要模拟 加载，我们将使用表达人源化野生型抗体而没有FAD突变的Ki小鼠(ApphAβWT/haβWT；参考 β小鼠)、人载脂蛋白E亚型(载脂蛋白E_2/E_2、载脂蛋白E_3/E_3和载脂蛋白E_4/E_4；E_2、E_3和E_4鼠)和人 野生型tau(MAPTWT/WT，TAUWT小鼠)。为了模拟时尚，我们将使用表达人性化野生型的Ki小鼠 具有瑞典和伊比利亚FAD突变(AppNL-F/NL-F；↑抗体小鼠)的AB，人类apoE亚型(E3和E4)，以及 人类野生型tau(TAUWT)。因此，我们建议研究人类的生理和内源性相互作用。 AB、apoE亚型和tau在体内导致AD相关神经网络活动、认知功能异常 使用最先进的活体电生理学、光遗传学、 和行为方法。我们将专注于改变神经网络功能障碍的机制，因为它们 阿尔茨海默病与大脑和认知功能密切相关，在阿尔茨海默病发病早期就被破坏。 在目标1中，我们将确定导致神经改变的抗体、apoE亚型和tau的致病相互作用。 LOAD和FAD KI小鼠疾病进展过程中的网络活动(目标1a)和行为缺陷(目标1b) 使用无线长期脑电/肌电记录以及标准和机器学习行为方法。在……里面 目的2，我们将确定抗体和载脂蛋白E4对细胞和电路功能的致病作用 LOAD和FAD KI小鼠体内的损伤(目标2a，b)和特定中间神经元的光遗传激活 细胞类型逆转AD相关的异常(目标2c)。我们将使用活体LFP和多单元记录和 行为小鼠识别细胞类型和网络电路级机制的光遗传学方法 功能障碍。在目标3中，我们将确定抗体、载脂蛋白E亚型和tau的致病相互作用。 Load和Fad Ki小鼠的AD相关病理(Aim 3a)和scRNA-seq转录组变化(Aim 3b)。我们 将在具有功能特征的小鼠中进行与AD相关的病理和scRNA-seq转录分析 (体内生理学和行为)从AIMS 1和2识别病理和转录组变化 与认知脆弱性和恢复力有关。我们的目标是识别经过验证的人类和行为 AD发病机制的相关候选基因/途径，并确定特定细胞类型的疾病机制。