Multidimensional mapping of vulnerable cell types in humanized Alzheimer's disease mouse models

人性化阿尔茨海默病小鼠模型中脆弱细胞类型的多维图谱

• 批准号: 10667216
• 负责人: Inma Cobos
• 金额: $ 235.44万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2028-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10667216
• 关键词: ATAC-seq; Aging; Alzheimer' s Disease; Alzheimer' s disease brain; Alzheimer' s disease model; Alzheimer' s disease pathology; Alzheimer' s disease related dementia; Amygdaloid structure; Amyloid; Amyloid beta-Protein; Astrocytes; Axon; Bar Codes; Behavior; Behavioral; Brain; Brain region; Cell Communication; Cell Nucleus; Cells; Cessation of life; Chronic; Cognitive deficits; Custom; Data; Data Set; Dimensions; Disease; Disease Progression; Early identification; Electroencephalography; Electrophysiology (science); Genes; Genetic Transcription; Goals; Haplotypes; Heterogeneity; Hippocampus; Human; Impairment; Inflammation; Interneurons; Knock-in; Knock-in Mouse; Limbic System; Machine Learning; Maps; Methods; Microglia; Modeling; Molecular; Morphology; Motor Cortex; Multiomic Data; Mus; Mutation; Neocortex; Nerve Degeneration; Neurons; Oligodendroglia; Pathogenicity; Pathology; Physiology; Population; Predisposition; Prefrontal Cortex; Property; Proteomics; Proto-Oncogene Protein c-kit; Receptor Protein-Tyrosine Kinases; Research; Resistance; Resolution; Senile Plaques; Sex Differences; Somatosensory Cortex; Synapses; Tauopathies; Testing; Visual Cortex; Work; abeta accumulation; behavioral phenotyping; behavioral study; brain cell; cell type; epigenome; epigenomics; familial Alzheimer disease; human data; human disease; humanized mouse; in vivo; mouse model; multiple omics; nano-string; neocortical; network dysfunction; neurofibrillary tangle formation; neuronal cell body; novel; optogenetics; presynaptic; progressive neurodegeneration; protein aggregation; protein expression; response; single-cell RNA sequencing; tau Proteins; tau aggregation; transcriptome; transcriptomics

ABSTRACT Alzheimer’s disease (AD) is characterized by progressive neurodegeneration and the aggregation of amyloid-b (Ab) and tau. The selective vulnerability of different brain regions and some cell types to AD pathology has been established. However, much remains unknown regarding the disease-relevant mechanisms underlying this differential response. We have previously used single-cell transcriptomics to perform an unbiased characterization of vulnerable and resistant neuronal subtypes in the human AD brain (19 excitatory and 24 inhibitory subtypes; ~490,000 nuclei, multi-region dataset). This characterization revealed early transcriptional changes in inhibitory interneurons, particularly in a population expressing the receptor tyrosine kinase c-Kit. Using our novel method to isolate by FACS and profile neuronal somas with tau aggregates, we also quantified the susceptibility of 20 major neocortical neuronal subtypes to the formation of neurofibrillary tangles (NFTs). Although interneurons proved generally resistant to NFT formation, they were not spared from death. Our work in the human brain highlights the existence of shared and distinct Ab- and tau-associated pathogenic mechanisms as well as the need for a multidimensional approach to characterizing vulnerability in AD. This proposal seeks to further characterize cell type-specific signatures of vulnerability to Ab and tau proteinopathies in newly developed knock-in (KI) humanized mouse models of AD. We will test the hypothesis that early changes in specific populations of GABAergic inhibitory interneurons, including c-Kit cells, are associated with network dysfunction, early protein aggregation, and cognitive deficits in humanized AD mouse models. To model pathogenic interactions between Ab and tau, we will use mouse models expressing humanized Ab without or with familial AD (FAD) mutations and mouse models expressing human MAPT without or with a mutation associated with tauopathy. In Aim 1, we will apply combined single-cell RNA- and ATAC-seq to tau-bearing and tau-free somas from mice characterized behaviorally and electrophysiologically by chronic EEG/EMG recordings and by standard and machine learning-analyzed behavior. In Aim 2, we will use spatial multiomics with single- cell and subcellular resolution to map cell-type-specific vulnerabilities and cell-cell interactions in relation to Ab and tau proteinopathies. In Aim 3, we will determine if Ab and/or tau alter the molecular, cellular, and circuit properties of vulnerable c-Kit interneurons. In all aims, we will integrate our multiomics and functional data and compare our mouse and previously-generated human data to identify evolutionarily conserved or species- specific cell type behaviors. The completion of these aims will provide a human disease-relevant, large-scale multiomics dataset instrumental to unravelling the mechanisms of neurodegeneration associated with Ab and tau proteinopathies.

抽象的 阿尔茨海默氏病（AD）的特征是进行性神经变性和淀粉样蛋白B的聚集 （AB）和Tau。不同大脑区域的选择性脆弱性和某些细胞类型已成为AD病理学 已确立的。但是，关于这一点的基于疾病的机制，尚不清楚 差异响应。我们以前已经使用了单细胞转录组学来执行公正 人类AD大脑中脆弱和抗性神经元亚型的表征（19兴和24 抑制亚型； 〜490,000个核，多区域数据集）。这种特征揭示了早期转录 抑制性神经元的变化，特别是在表达受体酪氨酸激酶C-KIT的种群中。 使用我们的新方法通过FACS和轮廓神经元与Tau聚集体分离，我们还量化了 20个主要的新皮质神经元亚型对神经原纤维缠结（NFTS）形成的敏感性。 尽管中间神经元通常证明对NFT形成具有抵抗力，但它们并未免于死亡。我们的工作 在人的大脑中，强调了共享和独特的AB和TAU相关的致病性的存在 机制以及对表征AD脆弱性的多维方法的需求。这 提案旨在进一步表征针对AB和TAU蛋白质病的脆弱性特异性特异 在新开发的AD的敲入（KI）人源化小鼠模型中。我们将测试早期变化的假设 在特定的GABA能抑制性中间神经元（包括C-KIT细胞）中，网络与网络有关 功能障碍，早期蛋白质聚集和认知在人源化AD小鼠模型中定义。建模 AB和Tau之间的致病相互作用，我们将使用表达人性化AB的小鼠模型，而无需或 使用家庭AD（FAD）突变和小鼠模型表达人类的MATT，而无需或具有突变 与tauopathy有关。在AIM 1中，我们将将组合的单细胞RNA-和ATAC-SEQ应用于tau-bearing和 小鼠的无tau somas在行为和电生理上以慢性脑电图/EMG记录在行为和电生理上表征 并按照标准和机器学习分析的行为。在AIM 2中，我们将使用单个空间多组学 与AB相关的细胞和亚细胞分辨率 和tau蛋白质。在AIM 3中，我们将确定AB和/或TAU是否改变了分子，细胞和电路 弱势c-kit中间神经元的特性。总体而言，我们将整合我们的多组学和功能数据，以及 比较我们的小鼠和先前生成的人类数据，以识别进化构成或物种 - 特定的细胞类型行为。这些目标的完成将提供与人类疾病相关的大规模 多组学数据集有助于阐明与AB和AB和 tau蛋白质。