Glycosylated clusterin species facilitate amyloid beta toxicity in human neurons.

Clusterin (CLU) is one of the most significant genetic risk factors for late onset Alzheimer's disease. Numerous studies have now demonstrated that CLU-AD mutations and amyloid-ß (Aß) treatment alter the trafficking and localisation of glycosylated CLU. iPSCs with altered CLU trafficking were generated following the removal of CLU exon 2 by CRISPR/Cas9 gene editing. Neurons were generated from control, unedited and exon 2 -/- iPSCs and were incubated with aggregated Aß peptides. Changes in cell death and neurite length were quantified to determine if altered CLU protein trafficking influenced neuronal sensitivity to Aß. Overall design: RNA-Seq analysis was performed to identify key transcriptomic differences between exon 2 -/- and CTR neurons.

簇蛋白（Clusterin, CLU）是晚发性阿尔茨海默病最关键的遗传风险因子之一。目前已有大量研究证实，CLU-AD相关突变以及淀粉样β（amyloid-β, Aβ）处理会改变糖基化CLU的运输与定位。本研究通过CRISPR/Cas9基因编辑技术敲除CLU外显子2，成功构建了CLU运输功能异常的诱导多能干细胞（induced pluripotent stem cells, iPSCs）。随后，分别从野生型对照、未编辑以及CLU外显子2纯合敲除的iPSCs中分化获得神经元，并将其与聚集态Aβ肽共孵育。通过量化细胞死亡与神经突长度的变化，以明确CLU蛋白运输异常是否会影响神经元对Aβ的敏感性。整体实验设计：本研究通过RNA测序（RNA-Seq）分析，鉴定出CLU外显子2纯合敲除神经元与对照神经元之间的关键转录组差异。