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

项目概要/摘要 AD是一种与年龄相关的神经退行性疾病，影响10%的65岁以上人口。遗传学晚期- AD（LOAD）是一种复杂的疾病。多个常见变量通常影响LOAD，但效应量较小。 这些预测的AD风险基因是否以及如何促进AD发病机制，以及哪些变体影响其 在大多数情况下，表达仍有待定性。拟议研究的总体目标是 为AD风险基因的作用提供全面的注释，并确定AD偶然变异有助于 AD通过调节AD风险基因表达。我们将：（1）确定和基准的生物后果 通过使用CRISPRi和单细胞RNA-seq在iPSC来源的细胞中进行遗传分析， 兴奋性神经元、海马齿状颗粒细胞和小胶质细胞。我们还将使用函数 在细胞和类器官中测定AD相关表型，（2）表征增强子和AD相关变体 AD风险基因在iPSC衍生的海马DG神经元和海马类器官中的作用，（3）阐明 iPSC衍生的兴奋性神经元、小胶质细胞和脑类器官中的增强子和AD变体， 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。 影响基因表达网络和与AD发病机制相关的细胞功能的因果变异。的 我们的建议的成功将是朝着更好的风险预测和新的治疗靶点迈出的重要一步。 AD.

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.