Elucidate the roles of Alzheimer's disease risk genes and variants in gene expression and AD-related phenotypes

项目摘要/摘要 AD是一种与年龄相关的神经退行性疾病，影响着65岁以上人口的10%。后来的遗传学- 发病AD(负荷)是复杂的。多个常见变量通常以较小的效果大小影响载荷。 这些预测的AD风险基因是否以及如何在AD发病中起作用，以及哪些变异会影响它们的 表情在很大程度上仍有待于定性。拟议研究的首要目标是 为AD风险基因的角色提供全面的注释，并确定AD偶然变量对 AD通过调节AD风险基因的表达而发挥作用。我们将：(1)确定生物后果并制定基准 应用CRISPRi和单细胞RNA-seq对IPSC来源的AD风险基因进行遗传分析 兴奋性神经元、海马齿状颗粒细胞和小胶质细胞。我们还将使用Function 细胞和器官中AD相关表型的分析，(2)增强剂和AD相关变异体的特征 对于IPSC来源的海马DG神经元和海马器类中的AD危险基因，(3)阐明其作用 IPSC来源的兴奋性神经元、小胶质细胞和脑器官中增强剂和AD变体的研究 小胶质细胞。拟议的工作将提供AD风险基因的全面注释，并揭开AD的神秘面纱。 影响阿尔茨海默病发病相关基因表达网络和细胞功能的因果变异。这个 我们建议的成功将是朝着更好的风险预测和新的治疗目标迈出的重要一步 广告。

Project Summary/Abstract AD is an age-related neurodegenerative disorder affecting 10% of the population over 65. The genetics of Late- onset AD (LOAD) is complex. Multiple common variants usually influence LOAD with smaller effect sizes. Whether and how these predicted AD risk genes contribute to AD pathogenesis and which variant affect their expression remain, for the most part, to be characterized. The overarching goal of the proposed study is to provide a comprehensive annotation of roles for AD risk genes and determine AD casual variants contribute to AD via modulating AD-risk gene expression. We will: (1) determine and benchmark the biological consequences of putative AD risk genes identified by genetic analysis using CRISPRi and single-cell RNA-seq in iPSC-derived excitatory neurons, hippocampal dentate granule cells, and microglia. We will also follow up using functional assays in cells and organoids for AD-related phenotypes, (2) characterize enhancers and AD-associated variants for AD risk genes in iPSC-derived hippocampal DG neurons and hippocampal organoids, (3) elucidate the roles of enhancers and AD variants in iPSC-derived excitatory neurons, microglia, and cerebral organoids with microglia. The proposed work will provide a comprehensive annotation of AD risk genes and demystify AD- causal variants affecting gene expression networks and cellular functions related to AD pathogenesis. The success of our proposal will be an essential step towards better risk prediction and new therapeutic targets in AD.