Resolving selective vulnerability and disease progression in human Alzheimer's brain via single-cell RNA-seq

通过单细胞 RNA-seq 解决人类阿尔茨海默病大脑的选择性脆弱性和疾病进展

• 批准号: 10407487
• 负责人: Inma Cobos
• 金额: $ 39.13万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-15 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10407487
• 关键词: Affect; Age; Alzheimer' s Disease; Alzheimer' s disease brain; Alzheimer' s disease pathology; Alzheimer' s disease therapy; Astrocytes; Autopsy; Biological; Biological Assay; Blood Vessels; Brain; Brain region; Cell Death; Cell Nucleus; Cells; Cerebral cortex; Cerebrum; Cognitive deficits; Collection; Communities; Custom; Data; Detection; Development; Disease; Disease Progression; Disease model; Event; Formalin; Freezing; Functional disorder; Gene Expression; Gene Expression Profiling; Human; Individual; Lead; Link; Mediating; Memory Loss; Methods; Microfluidics; Microglia; Molecular; Mus; Nerve Degeneration; Neurofibrillary Tangles; Neuroglia; Neurons; Oligodendroglia; Paraffin Embedding; Pathogenesis; Pathogenicity; Pathologic; Pathology; Pathway interactions; Predisposition; Publishing; Reproducibility; Research; Resolution; Resources; Sampling; Small Nuclear RNA; Tauopathies; Technology; Testing; Thalamic structure; Time; Tissue-Specific Gene Expression; Tissues; Transcript; United States National Institutes of Health; area striata; base; cell type; cohort; cost effective; design; disorder control; drug discovery; effective therapy; human RNA sequencing; human data; human tissue; hyperphosphorylated tau; improved; innovative technologies; insight; nano-string; neuroimaging; neuropathology; novel; novel strategies; novel therapeutics; sensory cortex; single-cell RNA sequencing; symptomatic improvement; tau Proteins; tau aggregation; tau-1; transcriptome; transcriptome sequencing; transcriptomics

ABSTRACT Effective therapies for Alzheimer’s disease (AD) are urgently needed. The existing approaches have largely failed to improve symptoms or modify disease progression, in part because of our incomplete understanding of the disease pathogenesis. Previous neuropathology and neuroimaging studies have been instrumental in establishing the histopathological hallmarks of AD and the involvement of specific brain regions. However, the selective vulnerability of distinct neuronal and non-neuronal cell types to AD and the underlying molecular mechanisms remain largely unknown. We hypothesize that subpopulations of neuronal and non-neuronal cell types contribute distinctively to AD and propose the use of single-cell transcriptomics in human brain to identify these cell types and the underlying molecular mechanisms leading to AD pathology. Although applying this innovative technology to human brain is challenging, human AD brain is crucial for understanding the contributions of distinct cell types to disease, identifying the earliest pathogenic events, uncovering neuroprotective pathways, and defining the spread of pathology. We will study early- and late-affected cerebral cortical regions, including entorhinal, association, and primary sensory cortices, from subjects encompassing the full spectrum of disease progression (Braak stages I–VI) and age-matched healthy controls. This strategy will provide a comprehensive landscape of the vulnerable and resilient cell types, their transcriptome changes, and the spread of changes over time and across cortical regions. In Aim 1 we will use single-nucleus RNA- sequencing for the unbiased identification and transcriptome profiling of neurons, glia (microglia, astrocytes, and oligodendrocytes), and blood vessel cells. In Aim 2 we will test the hypothesis that distinct molecularly defined subpopulations of cortico-cortical and cortico-thalamic projection neurons are selectively vulnerable to degeneration. In Aim 3 we will examine the molecular changes associated with tau pathology using our newly developed assay for purifying and profiling single neurons bearing neurofibrillary tangles. These studies will generate the first single-cell transcriptome profiling of human neurons with tangles and provide insight into the tau-mediated mechanisms of neurodegeneration. Together, our studies will provide an unbiased and robust identification of the vulnerable and resilient cell types in AD and insight into the molecular mechanisms underlying the selective vulnerabilities. These data will provide a valuable resource to the scientific community for improved cell-type-based disease modeling and drug discovery.

抽象的 迫切需要针对阿尔茨海默病（AD）的有效疗法。现有的方法很大程度上 未能改善症状或改变疾病进展，部分原因是我们对 本病发病机制。先前的神经病理学和神经影像学研究有助于 建立 AD 的组织病理学标志和特定大脑区域的参与。然而， 不同神经元和非神经元细胞类型对 AD 的选择性脆弱性及其潜在分子 机制仍然很大程度上未知。我们假设神经元和非神经元细胞亚群 类型对 AD 有独特的贡献，并建议在人脑中使用单细胞转录组学来识别 这些细胞类型和导致 AD 病理的潜在分子机制。虽然应用这个 创新技术对人类大脑具有挑战性，人类AD大脑对于理解AD至关重要 不同细胞类型对疾病的贡献，识别最早的致病事件，揭示 神经保护途径，并定义病理学的传播。我们将研究早期和晚期受影响的大脑 皮质区域，包括内嗅皮质、关联皮质和初级感觉皮质，来自受试者 全面的疾病进展（Braak I-VI 期）和年龄匹配的健康对照。这个策略 将提供脆弱和有弹性的细胞类型及其转录组变化的全面概况， 以及变化随时间的推移和跨皮质区域的传播。在目标 1 中，我们将使用单核 RNA- 对神经元、神经胶质细胞（小胶质细胞、星形胶质细胞和星形胶质细胞）进行公正的鉴定和转录组分析的测序 少突胶质细胞）和血管细胞。在目标 2 中，我们将检验以下假设：不同的分子定义 皮质-皮质和皮质-丘脑投射神经元的亚群选择性地易受 退化。在目标 3 中，我们将使用我们的新方法来检查与 tau 病理学相关的分子变化。 开发了纯化和分析带有神经原纤维缠结的单个神经元的分析方法。这些研究将 生成第一个人类神经元缠结的单细胞转录组分析，并提供对缠结的深入了解 tau 介导的神经变性机制。总之，我们的研究将提供公正和稳健的 识别 AD 中脆弱和有弹性的细胞类型并深入了解潜在的分子机制 选择性漏洞。这些数据将为科学界提供宝贵的资源，以改进 基于细胞类型的疾病建模和药物发现。