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: 10461843
• 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/10461843
• 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; 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; wireless

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 – 总结 大量证据表明，人类淀粉样蛋白前体蛋白/淀粉样蛋白 b (APP/Ab)、apoE4 和 微管相关蛋白 tau 会导致阿尔茨海默病 (AD)，但它们的致病相互作用是 很大程度上不为人知。减少大脑中抗体的积累被认为是最合理的治疗方法 AD 的策略，但这种方法的多项临床试验都失败了，这表明 AD 的病理生理学 AD 比预期复杂得多，AD 相关蛋白的致病相互作用需要 得到更好的理解。为了在生理表达水平上解码 AD 的多因素病因，我们将 研究新开发的迟发性 (LOAD) 和家族性 (FAD) AD 敲入 (KI) 小鼠模型，无需 转基因过度表达，重点关注 Ab、apoE 和 tau 之间的致病相互作用。模拟 LOAD，我们将使用表达人源化野生型抗体且无 FAD 突变的 KI 小鼠（ApphAβWT/hAβWT；称为 Aβ 小鼠）、人 apoE 同工型（APOEE2/E2、APOEE3/E3 和 APOEE4/E4；E2、E3 和 E4 小鼠）和人 野生型 tau（MAPTWT/WT、TAUWT 小鼠）。为了模拟 FAD，我们将使用表达人源化野生型的 KI 小鼠 具有瑞典和伊比利亚 FAD 突变的抗体（AppNL-F/NL-F；↑Ab 小鼠）、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 亚型和体内 tau 蛋白会导致 AD 相关的神经网络活动、认知异常 使用最先进的体内电生理学、光遗传学、 和行为方法。我们将重点关注改变神经网络功能障碍的机制，因为它们 与大脑和认知功能密切相关，并且在 AD 发病早期就被破坏。 在目标 1 中，我们将确定 Ab、apoE 同工型和 tau 的致病性相互作用，从而导致神经改变 LOAD 和 FAD KI 小鼠在疾病进展期间的网络活动（目标 1a）和行为缺陷（目标 1b） 使用无线长期脑电图/肌电图记录以及标准和机器学习行为方法。在 目标 2，我们将确定 Ab 和 apoE4 对细胞和回路功能的致病相互作用 LOAD 和 FAD KI 小鼠体内损伤（目标 2a、b）以及特定中间神经元的光遗传学激活 细胞类型可逆转 AD 相关异常（目标 2c）。我们将使用体内 LFP 和多单元录音 行为小鼠的光遗传学方法可识别细胞类型和网络的电路级机制 功能障碍。在目标 3 中，我们将确定 Ab、apoE 同工型和 tau 的致病相互作用，从而导致 LOAD 和 FAD KI 小鼠中 AD 相关病理学 (Aim 3a) 和 scRNA-seq 转录组变化 (Aim 3b)。我们 将在功能特征小鼠中进行 AD 相关病理和 scRNA-seq 转录组学分析 （体内生理学和行为）从目标 1 和 2 识别病理和转录组变化 与认知脆弱性和复原力有关。我们的目标是识别人类经过验证和行为的 AD 发病机制的相关候选基因/途径并定义细胞类型特异性疾病机制。