Characterizing the landscape of cell-type specific changes associated with Alzheimer's disease before death with single-cell genomics

用单细胞基因组学描述死亡前与阿尔茨海默病相关的细胞类型特异性变化的情况

• 批准号: 10680438
• 负责人: Tushar Vinod Kamath
• 金额: $ 5.04万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10680438
• 关键词: Alleles; Alzheimer' s Disease; Alzheimer' s disease brain; Alzheimer' s disease model; Alzheimer' s disease pathology; Alzheimer' s disease risk; Amyloid; Autopsy; Benchmarking; Biopsy; Biopsy Specimen; Brain; Brain Pathology; Cause of Death; Cell Nucleus; Cells; Cellular Assay; Cessation of life; Complex; Data Set; Disease; Fluorescent in Situ Hybridization; Freezing; Gene Expression; Genetic Transcription; Genetic study; Genomics; Heritability; Human; Immunohistochemistry; Impaired cognition; In Situ; Individual; Location; Measures; Memory Loss; Methods; Modeling; Molecular; Molecular Profiling; Morphologic artifacts; Neurofibrillary Tangles; Neuroimmune; Normal Pressure Hydrocephalus; Pathogenesis; Pathologic; Pathology; Pathway interactions; Patients; Population; Process; Protocols documentation; Regression Analysis; Research; Resolution; Resources; Role; Sampling; Senile Plaques; Signal Transduction; Slice; Slide; Techniques; Technology; Tissues; Transcription Alteration; Transgenic Mice; Variant; Work; brain tissue; cell type; cohort; comorbidity; computer framework; differential expression; end stage disease; experimental study; familial Alzheimer disease; frontal lobe; genome-wide; human tissue; insight; meter; mouse model; novel; response; single cell sequencing; single cell technology; single molecule; single nucleus RNA-sequencing; tau Proteins; tau-1; tool

Project Summary Alzheimer’s disease (AD) remains the only illness in the top 10 causes of death with no disease-modifying treatments available. In large part, the dearth of adequate therapies is due to our incomplete understanding of how specific molecular pathways in the brain result in cognitive decline and memory loss. Recent genetic studies implicate multiple neuroimmune populations in the brain as central players in the pathogenesis of AD, yet the precise molecular roles of these cell types remain largely unclear. Advances in single-cell technology have opened up the ability to robustly assay cell states within complex tissues, including the human brain. The ability to measure the precise cellular states in the living brain is key to understanding subtle neuroimmune cell type transcriptional changes that may give rise to AD. To identify molecular changes in these cells in human AD brain tissue, we have deployed single-nuclei RNA-sequencing (snRNA-seq) to profile frontal cortex biopsies from patients with suspected idiopathic normal pressure hydrocephalus (iNPH) and co-morbid AD. These biopsies represent a unique opportunity to measure transcriptional changes associated with AD unconfounded by postmortem artifact and, potentially, prior to the end stage of disease. To date, I have successfully obtained 815,843 single-nuclei profiles from 18 individuals, six of whom have both amyloid and tau pathology, seven with amyloid plaques, and five with no pathology. Here, I propose to employ snRNA-seq and Slide-seq on a total of 62 frontal cortex biopsies, across a range of APOE and amyloid/tau statuses, to uncover molecular alterations specifically associated with AD. Initially, I will determine which cell populations are significantly enriched or depleted in AD pathology and associated with APOE status, identify transcriptional alterations in all cell types, and determine which populations are enriched for heritable risk of AD. With Slide- seq, I will determine how cellular states and transcriptional changes are influenced by the spatial location in relation to amyloid and tau pathology. Finally, using novel computational integration methods, I will benchmark the transcriptional changes seen in a mouse model of AD with those obtained from my human profiling efforts, providing the field a resource of cellular state changes that are recapitulated in this model. If successful, these experiments will provide a comprehensive view of AD before death, nominating new cell state changes and transcriptional pathways associated with the pathogenesis of AD.

项目摘要 阿尔茨海默氏病（AD）仍然是死亡的十大原因中唯一的疾病，没有疾病修改 可用的治疗。在很大程度上，适当疗法的死亡是由于我们对 大脑中的特定分子途径如何导致认知能力下降和记忆力丧失。最近的遗传 研究暗示大脑中的多个神经免疫种群是AD发病机理的中心参与者， 然而，这些细胞类型的精确分子作用在很大程度上仍然不清楚。单细胞技术的进步 已经开放了在包括人脑在内的复杂组织中强稳定的细胞态的能力。这 测量活大脑中精确细胞状态的能力是了解细微的神经免疫细胞的关键 类型转录变化可能导致AD。确定这些细胞中人类中的分子变化 广告脑组织，我们已部署了单核RNA-severing（SnRNA-Seq）到剖面额叶皮层 来自怀疑特发性正常压力脑积水（INPH）和合并AD患者的活检。 这些活检代表了测量与AD相关的转录变化的独特机会 在疾病终结阶段之前，不受尸体伪像的不符合。迄今为止，我有 成功获得了来自18个人的815,843个单核轮廓，其中6个具有淀粉样蛋白， tau病理学，七个带有淀粉样蛋白斑块，没有病理。在这里，我建议使用snrna-seq 以及在一系列APOE和淀粉样蛋白/TAU统计的总共62个额叶皮层活检中的幻灯片序列 发现与AD特别相关的分子改变。最初，我将确定哪些细胞群体 在AD病理学中显着丰富或耗尽，并与APOE状态相关，识别转录 所有细胞类型的改变，并确定哪些种群富含AD的遗传风险。幻灯片 - Seq，我将确定细胞状态和转录变化如何受到空间位置的影响 与淀粉样蛋白和TAU病理学有关。最后，使用新颖的计算集成方法，我将基准测试 从我的人类分析工作中获得的AD鼠标模型中看到的转录变化， 提供该模型中概括的蜂窝状态变化的资源。如果成功，这些 实验将在死亡前提供全面的广告视图，提名新的细胞状态变化，并 与AD发病机理相关的转录途径。