Mapping Cellular Resolution Connectopathies in Aging and Alzheimer's Disease

绘制衰老和阿尔茨海默氏病的细胞分辨率连接病图谱

• 批准号: 10621814
• 负责人: Hong-Wei Dong
• 金额: $ 288.84万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-15 至 2027-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10621814
• 关键词: 3-Dimensional; Affect; Age; Aging; Alzheimer' s Disease; Alzheimer' s disease model; Alzheimer' s disease pathology; Amyloid; Amyloid beta-Protein; Anatomy; Atlases; Axon; BRAIN initiative; Biological Assay; Cause of Death; Cell Nucleus; Cells; Chromatin; Collection; Computer software; Consensus; Data; Data Analyses; Dementia; Elderly; Female; Genetic; Genetic Models; In Situ Hybridization; Informatics; Knock-in; Knock-in Mouse; Label; Late Onset Alzheimer Disease; Maps; Modeling; Molecular; Molecular Profiling; Morphology; Mus; Neuroglia; Neurons; Online Systems; Output; Pathologic; Pathology; Pathway Analysis; Process; Production; Resolution; Resources; Source; Synapses; System; Techniques; Testing; United States; Viral; Visualization; age related; amyloid pathology; analysis pipeline; cell cortex; cell type; cloud based; connectome; connectome data; data portal; data visualization; digital; disease mechanisms study; effective therapy; entorhinal cortex; epigenomics; genome-wide; hippocampal pyramidal neuron; in vivo; male; methylomics; molecular phenotype; mouse genetics; mouse model; multidisciplinary; multimodality; multiple omics; mutant; neurotechnology; new technology; next generation; normal aging; novel; reconstruction; response; scale up; sex; shape analysis; single nucleus RNA-sequencing; tau Proteins; therapeutic candidate; transcriptomics

PROJECT SUMMARY/ABSTRACT The proposed project, “Mapping Cellular Resolution Connectopathies in Aging and Alzheimer's Disease,” will systematically and comprehensively characterize cell-type specific anatomical and molecular phenotypes across aging and Alzheimer’s disease (AD) in the entorhinal cortex (ENT): the ground zero of AD pathology. We will map cellular resolution, age-dependent morpho-molecular phenotypes of ENT projection neurons by performing single-nucleus RNA-sequencing and methylomic analysis in 2m, 9m, 18m APPSAA-(KI/KI) male and female mice. Novel genetic sparse labeling will be used to label and characterize morphology of ENT pyramidal neurons in different cortical layers in APPSAA-(KI/KI)/MORF3/Cux2-CreER and APPSAA-(KI/KI)/MORF3/Etv1-CreER mice. These studies will provide comprehensive data on how molecularly defined ENT neuronal cell types interact with age-, sex- and Aβ pathology to confer progressive transcriptomic/epigenomic, morphological, and synaptic deficits in vivo. In addition, we will map the age-dependent morpho-molecular phenotypes of ENT projection neurons in humanized Tau models, MAPT(H1)-GR*N279K and their MAPT(H1) controls. A combined single- nucleus transcriptomics and genome-wide chromatin accessibility assays will be applied to MAPT(H1)- GR*N279K and MAPT(H1) male and female mice at 2m, 9m, and 18m to define integrated transcriptomic/epigenomic ENT neuronal cell types, and to identify neuronal subsets undergoing age-dependent multi-modal molecular dysregulation in mutant “humanized” Tau mouse models. RNAscope multiplex in situ hybridization and GeoMX digital spatial profiling analyses will be performed to identify morpho-molecular types of neurons in ENT that are most affected in MAPT(H1)-GR*N279K knock-in mice compared to MAPT(H1) mice during aging. To identify age-related connectional vulnerabilities and to map connectivity disruptions in AD, we will systematically quantify changes of axonal outputs arising from genetically and connectionally defined ENT cell types using 2m, 9m, 18m male and female Cux2-CreER and Etv1-CreER mice. Cell-type specific connectivity disruptions also will be examined in two next generation AD mouse models, APP knock-in (APPSAA-KI/KI with wildtype controls) and MAPT(H1)-GR*N279K [with MAPT(H1) controls], across ages and in both sexes. Novel viral sparse labeling will be used to characterize age- and AD-related axonal dystrophy, while genetic sparse labeling in newly generated MORF3 mouse lines will help to identify local morphological changes in ENT cell types. Age- and AD-related morphological compromises to ENT input neurons will be studied along with their synaptic disruptions onto different ENT cell types. Finally, we will establish a cloud-based visualization platform to map the integrated molecular-anatomic circuit deficits of aging and AD to the Allen Common Coordinate Framework to facilitate dissemination and analysis of the data. Although the focus of the current project is the ENT, the pipelines established for data production, collection, and analysis can be scaled up to identify brainwide cell-type specific anatomic-molecular deficits in aging and other late-onset AD mouse models.

项目概要/摘要 拟议的项目“绘制衰老和阿尔茨海默病中的细胞分辨率连接病”将 系统地、全面地表征跨细胞类型的特定解剖学和分子表型 内嗅皮质 (ENT) 中的衰老和阿尔茨海默病 (AD)：AD 病理学的归零点。我们将 通过执行绘制耳鼻喉投射神经元的细胞分辨率、年龄依赖性形态分子表型 2m、9m、18m APPSAA-(KI/KI) 雄性和雌性小鼠的单核 RNA 测序和甲基组学分析。 新型遗传稀疏标记将用于标记和表征耳鼻喉锥体神经元的形态 APPSAA-(KI/KI)/MORF3/Cux2-CreER 和 APPSAA-(KI/KI)/MORF3/Etv1-CreER 小鼠的不同皮质层。 这些研究将提供关于分子定义的耳鼻喉神经元细胞类型如何与 年龄、性别和 Aβ 病理学赋予渐进的转录组/表观基因组、形态学和突触 体内缺陷。此外，我们将绘制 ENT 投影的年龄依赖性形态分子表型 人源化 Tau 模型、MAPT(H1)-GR*N279K 及其 MAPT(H1) 对照中的神经元。组合单 核转录组学和全基因组染色质可及性测定将应用于 MAPT(H1)- GR*N279K 和 MAPT(H1) 雄性和雌性小鼠在 2m、9m 和 18m 处定义积分 转录组/表观基因组耳鼻喉神经元细胞类型，并鉴定经历年龄依赖性的神经元亚群 突变“人源化”Tau 小鼠模型中的多模式分子失调。 RNAscope 多重原位分析 将进行杂交和 GeoMX 数字空间分析分析来识别形态分子类型 与 MAPT(H1) 小鼠相比，MAPT(H1)-GR*N279K 敲入小鼠中耳鼻喉科中受影响最严重的神经元的数量 老化期间。为了识别与年龄相关的连接漏洞并绘制 AD 中的连接中断图，我们 将系统地量化由遗传和连接定义的耳鼻喉科引起的轴突输出的变化 使用 2m、9m、18m 雄性和雌性 Cux2-CreER 和 Etv1-CreER 小鼠的细胞类型。细胞类型特定连接 还将在两个下一代 AD 小鼠模型、APP 敲入（APPSAA-KI/KI 与 野生型对照）和 MAPT(H1)-GR*N279K [与 MAPT(H1) 对照]，跨年龄和性别。小说 病毒稀疏标记将用于表征年龄和 AD 相关的轴突营养不良，而遗传稀疏标记将用于表征年龄和 AD 相关的轴突营养不良 新生成的 MORF3 小鼠系中的标记将有助于识别 ENT 细胞的局部形态变化 类型。将研究耳鼻喉输入神经元与年龄和 AD 相关的形态学妥协及其 不同耳鼻喉细胞类型的突触破坏。最后，我们将建立一个基于云的可视化平台 将衰老和 AD 的综合分子解剖回路缺陷映射到艾伦通用坐标 促进数据传播和分析的框架。虽然当前项目的重点是 耳鼻喉科，为数据生产、收集和分析而建立的管道可以扩大规模以识别全脑 衰老和其他晚发性 AD 小鼠模型中细胞类型特异性解剖分子缺陷。