Single-cell transcriptomic and epigenomic analysis of brain cell vulnerabilities to tauopathies in early AD impacted brain regions

早期 AD 影响大脑区域脑细胞对 tau 蛋白病变脆弱性的单细胞转录组和表观基因组分析

• 批准号: 10667016
• 负责人: MICHAEL D KOOB
• 金额: $ 209.45万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-01 至 2028-02-29
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10667016
• 关键词: ATAC-seq; Age; Aging; Alzheimer' s Disease; Alzheimer' s disease model; Alzheimer' s disease related dementia; Amyloid beta-Protein; Anatomy; ApoE knockout mouse; Apolipoprotein E; Atlases; Attenuated; Axon; Behavioral; Biological; Biological Assay; Brain; Brain Injuries; Brain region; Cell physiology; Cells; Chromatin; Chromium; Chromosome Mapping; Computer Analysis; DNA; Data; Deposition; Development; Disease; Epigenetic Process; Fluorescent in Situ Hybridization; Gene Expression; Genetic; Genomics; Goals; Hippocampus; Human; Image; Immediate-Early Genes; Individual; Institution; Joints; Knock-in; Literature; MAPT gene; Maps; Measures; Minnesota; Molecular; Molecular Profiling; Multiomic Data; Mus; Neurofibrillary Tangles; Neurons; Pathogenesis; Pathogenicity; Pathologic; Pathology; Phase; Phosphorylation; Physiological; Population; Predisposition; Protein Isoforms; Publishing; RNA; Research; Research Personnel; Resistance; Resolution; Resources; Senile Plaques; Spatial Distribution; Standardization; Tauopathies; Technology; Testing; Tissue Preservation; Transgenic Mice; Universities; Variant; Work; age related; apolipoprotein E-4; brain cell; cell type; cerebral atrophy; computational pipelines; diagnostic tool; entorhinal cortex; epigenomics; gene replacement; hyperphosphorylated tau; insight; locus ceruleus structure; mild traumatic brain injury; model development; molecular pathology; mouse model; multiple omics; neural circuit; neuromechanism; neuron loss; neuronal circuitry; neuronal excitability; neuropathology; next generation; novel; novel therapeutic intervention; response; tau Proteins; transcriptomics; validation studies

Project Summary Alzheimer's disease (AD) is an age-related, progressive neurogenerative disease that leads to loss of brain cells and their connections. The existing literature provides a broad view of AD/ADRD-related molecular changes from heterogeneous cell populations. However, cell-specific and brain-region specific AD/ADRD-related loss of cells and connectivity is not yet resolved, particularly at a mechanistic level. In response to RFA-AG-23-028, we have assembled a strong multi-investigator team across multiple institutions with complementary expertise in single-cell transcriptomics and epigenomics analysis, neural circuit mapping, and next-generation AD mouse model development. We will use multiple complementary lines of tau mouse models, in conjunction with APOE genetic modulation or pathogenic triggers in targeted brain regions. 1) We will use the Tau P301S transgenic mice on either a human APOE4 knock-in background (TE4) or a mouse Apoe knock-out background (TEKO). TE4 knock-in markedly exacerbates tau-mediated neurodegeneration, while TEKO mice show largely attenuated neuronal loss and brain atrophy compared to P301S mice. 2) We will additionally use recently developed novel humanized Tau mouse models that replace the endogenous mouse MAPT gene with either a normal or pathogenic variant of the entire human MAPT gene (MAPT gene replacement, MAPT-GR), which express all isoforms of human tau at physiologic levels and ratios. We hypothesize that vulnerable cell types in early AD- impact brain regions (locus coeruleus, entorhinal cortex, and hippocampal CA1 and subiculum) show early maladaptive gene expression profiles and epigenomic signatures that define their molecular vulnerability during AD/ADRD pathogenesis. To test our hypothesis, in Aim 1 we will apply single-cell epigenomics and transcriptomics technologies to early AD-impacted brain regions in age-matched control and AD mice, creating cell-type-resolved multi-omic maps of gene expression and chromatin accessibility. Tauopathy progression unfolds in an age-dependent manner, thus we will compare control and pathological tau model mice at two different ages each for different mouse lines (4 months, 10 months for TE and TEKO; 6 months, 12 months for MAPT-GR lines) based upon their behavioral and neuropathological characterization. In Aim 2, we will use the multiplexed error-robust fluorescence in situ hybridization (MERFISH) technology to generate single-cell resolution spatial transcriptomic maps for the early AD-impacted brain regions. MERFISH will extend single-cell omics and spatial genomics to map neural circuits and pathologies at high spatial resolution. In Aim 3, we will use perform computational analysis to integrate single-cell multi-omics data, image-based anatomical and molecular gene expression maps from Aims 1-2, acquired from different mouse models at different ages, to fully characterize the neuronal circuits and AD-vulnerability at cellular level. The proposed research is well aligned to the RFA goals and is expected to provide new biological insights into AD/ADRD pathogenesis at unprecedented cellular and spatial resolution.

项目摘要 阿尔茨海默氏病（AD）是一种与年龄相关的，进行性神经发生疾病，导致脑细胞丧失 和他们的联系。现有文献提供了与AD/ADRD相关的分子变化的广泛视图 来自异质细胞群。但是，细胞特异性和大脑区域特异性AD/ADRD相关的损失 细胞和连通性尚未解决，尤其是在机械水平上。为了响应RFA-AG-23-028，我们 已经组建了一个具有互补专业知识的多个机构的强大的多投资者团队 单细胞转录组学和表观基因组学分析，神经电路映射和下一代AD小鼠 模型开发。我们将使用tau鼠标模型的多个互补线和APOE结合 靶向大脑区域中的遗传调节或致病性触发因素。 1）我们将使用tau p301s转基因 在人APOE4敲门背景（TE4）或鼠标APOE敲除背景（TEKO）上的小鼠。 TE4敲入明显加剧了tau介导的神经变性，而Teko小鼠在很大程度上显示 与P301S小鼠相比，神经元丧失和脑萎缩。 2）我们还将使用最近开发的小说 人源化的tau小鼠模型，用正常或 整个人类MAPT基因（MAPT基因替代，MAPT-GR）的致病变异，它表达所有 人tau的同工型在生理水平和比率下。我们假设在早期的AD- 撞击大脑区域（院位，内嗅皮层和海马CA1和下图）显示早期 适应性基因表达谱和表观基因组特征，这些特征定义其分子脆弱性 AD/ADRD发病机理。为了检验我们的假设，在AIM 1中，我们将应用单细胞表观基因组学和 在年龄匹配的控制和AD小鼠中，转录组学技术到早期受ad侵入的大脑区域，创造 基因表达和染色质可及性的细胞类型分辨多摩尼图。 tauopathy的进展 以年龄的方式展开，因此我们将在两个 不同的老鼠线不同年龄（TE和TEKO的4个月，10个月； 6个月，12个月 mapt-gr线）基于其行为和神经病理学的特征。在AIM 2中，我们将使用 多路复用误差荧光原位杂交（Merfish）技术以生成单细胞 分辨率的空间转录组图，用于早期受ad影响的大脑区域。 Merfish将扩展单细胞 幻象和空间基因组学以高空间分辨率绘制神经回路和病理。在AIM 3中，我们将 使用执行计算分析以整合基于图像的解剖学和 来自AIMS 1-2的分子基因表达图，从不同年龄的不同小鼠模型获取到完全 表征神经元电路和细胞水平上的可质合性。拟议的研究非常顺利 达到RFA目标，并有望为AD/ADRD发病机理提供新的生物学见解 前所未有的细胞和空间分辨率。