The Role of ApoE in Neuroplasticity and A-beta; Clearance using iPS Cell-Derived Astrocytes

项目3：载脂蛋白E在iPS细胞来源的星形胶质细胞的神经可塑性和A？清除中的作用 项目摘要/摘要 这项建议旨在利用患者iPS细胞来源的星形胶质细胞来研究载脂蛋白E的作用 (APOE)在分子水平上对AD风险的影响。虽然小鼠模型极大地提高了我们对AD的理解 载脂蛋白E影响人类神经功能的分子和细胞机制及发病机制 退化在很大程度上仍是未知的。ADRC患者来源的星形胶质细胞将用于确定APOE 基因型影响星形胶质细胞的功能，并使用一种新型的人类星形胶质细胞/神经元共培养，确定其作用 载脂蛋白E亚型对突触形成/可塑性的影响，并研究突触对A和tau的易感性 受到了影响。最近的研究表明，载脂蛋白E也可能影响星形胶质细胞介导的A？清除。我们会 因此，确定不同的载脂蛋白E亚型是否改变了人类星形胶质细胞吞噬A的速率 并影响tau蛋白的神经元-神经元传递。拟议的研究将利用12个系列的患者- 由ADRC iPS细胞核心提供的衍生iPS细胞(6-APOE？3/？3和6-APOE？4/？4)。与 在iPS细胞核心，我们还将使用CRISPR介导的基因组编辑来转化APOE？4/？4 iPS细胞系 以使我们能够比较不同基因同源的人类星形胶质细胞 只有载脂蛋白E等位基因。最后，iPS细胞来源的星形胶质细胞将被移植到异种移植中- 亲和的Rag-5xfAD和Rag-tau小鼠，以检测apoE对体内淀粉样蛋白和tau病理的影响。

Project 3: The Role of ApoE in Neuroplasticity and A¿ Clearance using iPS Cell-Derived Astrocytes Project Summary/Abstract This proposal aims to utilize patient iPS cell-derived astrocytes to investigate the role of apolipoprotein E (apoE) on AD risk at the molecular level. While mouse models have greatly improved our understanding of AD pathogenesis, the molecular and cellular mechanisms by which apoE influences human neuronal function and degeneration remain largely unknown. ADRC patient-derived astrocyte will be used to determine how APOE genotype affects astrocyte function and using a novel human astrocyte/neuronal co-culture, determine the role of apoE isoforms on synapse formation/plasticity and to investigate how synaptic vulnerability to A¿ and tau is influenced. Recent studies suggest that apoE may also influence astrocyte-mediated A¿ clearance. We will therefore determine whether the different apoE isoforms alter the rate of A¿ phagocytosis by human astrocytes and influence the neuron-neuron transmission of tau. The proposed studies will utilize 12 lines of patient- derived iPS cells provided by the ADRC iPS Cell Core (6-APOE ¿3/¿3 and 6-APOE ¿4/¿4). In collaboration with the iPS Cell Core, we will also use CRISPR-mediated genome editing to convert an APOE ¿4/¿4 iPS cell line into an isogenic APOE ¿3/¿3 line to allow us to compare genetically homogenous human astrocytes that differs only by the APOE allele. Lastly, iPS cell-derived astrocytes will be transplanted into xenotransplantation- compatible Rag-5xfAD and Rag-tau mice to examine the effects of apoE on amyloid and tau pathology in vivo.