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

基本信息

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
项目类别：
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&apos s Disease Alzheimer&apos s disease model Alzheimer&apos 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

项目摘要

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.

项目摘要/摘要拟议的项目“绘制衰老和阿尔茨海默氏病中的细胞分辨率连接路径”，将会系统，全面地表征细胞类型的特定解剖学和分子表型内嗅皮层（ENT）中的衰老和阿尔茨海默氏病（AD）：AD病理的零地面。我们将 MAP细胞分辨率，通过执行ENT投影神经元的年龄依赖性形态分子表型在2M，9M，18M AppSAA-（Ki/ki）雄性和雌性小鼠中进行单核RNA测序和甲基分析。新型的遗传稀疏标记将用于标记和表征ENT金字塔神经元的形态 AppSAA-（KI/KI）/MORF3/CUX2-CREER和APPSAA-（KI/KI）/MORF3/ETV1-CREER小鼠中的不同皮层。这些研究将提供有关分子定义的ENT神经元细胞类型如何相互作用的全面数据年龄，性别和Aβ病理学以赋予进行性转录组/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模型中的神经元MAPT（H1）-gr*N279K及其MAPT（H1）对照。一个组合的单个细胞核转录组学和全基因组染色质可及性测定将用于MAPT（H1） - Gr*N279K和Mapt（H1）男性和雌性小鼠在2m，9m和18m处定义整合转录组/表观基因质ENT神经元细胞类型，并识别受年龄依赖性的神经元子集突变体“人性化” tau小鼠模型中的多模式分子失调。原位rnascope多路复用将进行杂交和GEOMX数字空间分析分析以识别形态分子类型与MAPT（H1）小鼠相比在衰老期间。为了识别与年龄相关的连接漏洞并绘制AD中的连接性破坏，我们将系统地量化由遗传和连接的ENT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。细胞类型的特定连接性在两个下一代AD鼠标模型，App knopk-In（AppSaa-ki/ki）中也将检查中断 WildType对照）和MAPT（H1）-gr*N279K [带有MAPT（H1）对照]，跨越年龄和男女。小说病毒稀疏标记将用于表征年龄和广告相关的轴突营养不良，而基因稀疏在新生成的MORF3鼠标线上的标记将有助于识别ENT细胞中的局部形态变化类型。年龄和广告相关的形态学对ENT输入神经元的妥协及其突触破坏不同的ENT细胞类型。最后，我们将建立一个基于云的可视化平台绘制综合分子 - 动态电路定义了衰老和AD到Allen common坐标促进数据传播和分析的框架。尽管当前项目的重点是 ENT，为数据生产，收集和分析建立的管道可以进行缩放以识别脑海细胞类型特异性解剖 - 分子在衰老和其他晚期AD小鼠模型中定义。