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.

项目概要 阿尔茨海默病 (AD) 是一种与年龄相关的进行性神经生成疾病，会导致脑细胞损失 以及他们的联系。现有文献提供了 AD/ADRD 相关分子变化的广泛观点 来自异质细胞群。然而，细胞特异性和脑区域特异性 AD/ADRD 相关的丧失 细胞和连接性尚未解决，特别是在机械层面。为了回应 RFA-AG-23-028，我们 已在多个机构组建了一支强大的多研究者团队，在以下领域具有互补的专业知识 单细胞转录组学和表观基因组学分析、神经回路图谱和下一代 AD 小鼠 模型开发。我们将使用多个互补系的 tau 小鼠模型，并结合 APOE 目标大脑区域的基因调节或致病触发因素。 1) 我们将使用 Tau P301S 转基因 人类 APOE4 敲入背景 (TE4) 或小鼠 Apoe 敲除背景 (TEKO) 的小鼠。 TE4 敲入显着加剧 tau 介导的神经变性，而 TEKO 小鼠则表现出很大程度上减弱 与 P301S 小鼠相比，神经元损失和脑萎缩。 2）我们将另外使用最近开发的小说 人源化 Tau 小鼠模型，将内源性小鼠 MAPT 基因替换为正常或 整个人类 MAPT 基因的致病性变异（MAPT 基因置换，MAPT-GR），表达所有 生理水平和比例的人类 tau 亚型。我们假设早期 AD 中的脆弱细胞类型 影响大脑区域（蓝斑、内嗅皮层、海马 CA1 和下托）早期显示 适应不良的基因表达谱和表观基因组特征定义了其分子脆弱性 AD/ADRD 发病机制。为了检验我们的假设，在目标 1 中，我们将应用单细胞表观基因组学和 转录组学技术对年龄匹配的对照小鼠和 AD 小鼠中早期受 AD 影响的大脑区域进行研究，创造了 细胞类型解析的基因表达和染色质可及性的多组学图谱。 tau蛋白病进展 以年龄依赖性方式展开，因此我们将在两个年龄时比较对照和病理性 tau 模型小鼠 不同的小鼠系各有不同的年龄（TE 和 TEKO 为 4 个月、10 个月；TE 和 TEKO 为 6 个月、12 个月） MAPT-GR 系）基于其行为和神经病理学特征。在目标 2 中，我们将使用 多重抗差错荧光原位杂交 (MERFISH) 技术生成单细胞 早期 AD 影响的大脑区域的分辨率空间转录组图。 MERFISH将扩展单细胞 组学和空间基因组学以高空间分辨率绘制神经回路和病理图。在目标 3 中，我们将 使用执行计算分析来整合单细胞多组学数据、基于图像的解剖学和 目标 1-2 的分子基因表达图谱是从不同年龄的不同小鼠模型中获得的，以充分 在细胞水平上表征神经元回路和 AD 脆弱性。拟议的研究非常一致 达到 RFA 目标，并有望为 AD/ADRD 发病机制提供新的生物学见解 前所未有的细胞和空间分辨率。