Study of Selective Cell and System Vulnerability in Alzheimer's Disease

项目摘要/摘要 阿尔茨海默病(AD)是老年人痴呆症的主要原因，最常见的年龄段- 与神经退行性疾病相关的疾病，影响全球约5600万人。尽管它的遗传性很高 (估计在60%-80%之间)和许多遗传变异(存在于整个基因组的数十个基因座上) 从全基因组关联研究(GWAS)中，我们对潜在的遗传机制的知识仍然存在 有限的。揭示阿尔茨海默病的病理生理机制被证明是非常具有挑战性的。至 推进我们对AD的机械性理解，我们将首先获取和协调各种内部、保护 和公开数据，包括散装和单细胞RNA-seq数据、全球气候变化综合统计数据、数组 基因分型和全基因组测序数据，以及功能基因组数据。然后我们将分析 他们使用一套计算方法和生物信息学工具来产生特定细胞类型 机械论假说。这些假设将通过实验技术得到验证。我们的 将在IPSC来源的神经细胞(特别是兴奋性神经元和小胶质细胞)中进行验证，因为 以及IPSC衍生的脑有机体，涉及不同类型的细胞，包括神经元、星形胶质细胞和 小胶质细胞。在这些IPSC派生的细胞和有机体模型中，我们将利用CRISPRi以及敲击 扰乱最有希望的假定因果调控DNA元件的实验技术，以及 通过测量包括基因表达在内的一系列分子和细胞表型来评估影响 AD相关生理表型。

PROJECT SUMMARY/ABSTRACT Alzheimer’s disease (AD) is a leading cause of dementia among the elderly and the most prevalent age- related neurodegenerative disease, affecting ~56 million individuals worldwide. Despite of its high heritability (estimates ranging 60-80%) and many genetic variants (residing in tens of loci across the genome) identified from genome-wide association studies (GWAS), our knowledge of the underlying genetic mechanisms remains limited. Uncovering pathophysiological mechanisms underlying AD proves to be highly challenging. To advance our mechanistic understanding of AD, we will first acquire and harmonize various in-house, protected and public data, encompassing bulk and single cell RNA-seq data, GWAS summary statistics, array genotyping and whole genome sequencing data, as well as functional genomic data. We will then analyze them using a suite of computational methods and bioinformatics tools to generate cell-type-specific mechanistic hypotheses. These hypotheses will be validated through experimental technologies. Our validations will be carried out in iPSC-derived neural cells (particularly excitatory neurons and microglia), as well as in iPSC-derived brain organoids involving diverse cell types including neurons, astrocytes, and microglia. In these iPSC-derived cells and organoids models, we will leverage CRISPRi as well as knock-in experimental technologies to perturb the most promising putatively causal regulatory DNA elements, and evaluate the impact by measuring a cascade of molecular and cellular phenotypes including gene expression and AD related physiological phenotypes.