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

基本信息

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

项目摘要

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.

抽象的阿尔茨海默病 (AD) 的特点是进行性神经变性和淀粉样蛋白 B 的聚集 (Ab) 和 tau 蛋白对 AD 病理学的不同大脑区域和某些细胞类型的选择性脆弱性进行了研究。然而，关于其背后的疾病相关机制仍不清楚。我们之前使用单细胞转录组学进行了公正的反应。人类 AD 大脑中脆弱和抵抗神经亚型的特征（19 种兴奋性和 24 种抑制亚型；~490,000 个细胞核，多区域数据集）。抑制性中间神经元的变化，特别是在表达受体酪氨酸激酶 c-Kit 的群体中。使用我们的新方法通过 FACS 进行分离并用 tau 聚集体分析神经体，我们还量化了 20 种主要新皮质神经元亚型对神经原纤维缠结 (NFT) 形成的易感性。尽管中间神经元普遍抵抗 NFT 的形成，但它们也未能幸免于我们的工作。人脑中的这一发现凸显了共同且独特的 Ab 和 tau 相关致病性的存在机制以及需要采用多维方法来描述 AD 中的脆弱性。该提案旨在进一步表征对 Ab 和 tau 蛋白病易感性的细胞类型特异性特征在新开发的 AD 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群体中，包括 c-Kit 细胞，与网络相关人源化 AD 小鼠模型中的功能障碍、早期蛋白质聚集和认知缺陷。 Ab 和 tau 之间的致病性相互作用，我们将使用表达人源化 Ab 的小鼠模型，而无需或具有家族性 AD (FAD) 突变的小鼠模型以及表达人类 MAPT（不带或带突变）的小鼠模型在目标 1 中，我们将结合单细胞 RNA 和 ATAC-seq 来研究 tau 蛋白携带和通过慢性脑电图/肌电图记录来表征小鼠的无 tau 体细胞的行为和电生理学特征根据标准和机器学习分析的行为，在目标 2 中，我们将使用空间多组学和单组学。细胞和亚细胞分辨率，以绘制与抗体相关的细胞类型特异性脆弱性和细胞间相互作用在目标 3 中，我们将确定 Ab 和/或 tau 蛋白是否会改变分子、细胞和回路。脆弱的 c-Kit 中间神经元的特性在所有目标中，我们将整合我们的多组学和功能数据并比较我们的小鼠和之前生成的人类数据，以确定进化上保守的或物种- 这些目标的完成将提供与人类疾病相关的大规模研究。多组学数据集有助于揭示与 Ab 和相关的神经变性机制 tau 蛋白病。