Amyotrophic lateral sclerosis overlapping with frontotemporal dementia (ALS/FTD) is a fatal and currently untreatable disease characterized by rapid cognitive decline and paralysis. Elucidating initial cellular pathologies is central to therapeutic target development, but obtaining samples from presymptomatic patients is not feasible. Here, we report the development of a cerebral organoid slice model derived from human induced pluripotent stem cells (iPSCs) that recapitulates mature cortical architecture and displays early molecular pathology of C9ORF72 ALS/FTD. Using a combination of single-cell RNA sequencing and biological assays, we reveal distinct transcriptional, proteostasis and DNA repair disturbances in astroglia and neurons. We show that astroglia display increased levels of the autophagy signaling protein P62 and that deep layer neurons accumulate dipeptide repeat protein poly(GA), DNA damage and undergo nuclear pyknosis that could be pharmacologically rescued by GSK2606414. Thus, patient-specific iPSC-derived cortical organoid slice cultures are a reproducible translational platform to investigate preclinical ALS/FTD mechanisms as well as novel therapeutic approaches.

与额颞叶痴呆 (ALS/FTD) 重叠的肌萎缩侧索硬化症是一种致命的、目前无法治愈的疾病，其特征是认知能力迅速下降和瘫痪。阐明最初的细胞病理学是治疗靶标开发的核心，但从症状前的患者身上获取样本是不可行的。在这里，我们报告了源自人类诱导多能干细胞 (iPSC) 的大脑类器官切片模型的开发，该模型概括了成熟的皮质结构并显示了C9ORF72的早期分子病理学肌萎缩侧索硬化症/FTD。我们结合使用单细胞 RNA 测序和生物测定，揭示了星形胶质细胞和神经元中明显的转录、蛋白质稳态和 DNA 修复障碍。我们发现星形胶质细胞显示自噬信号蛋白 P62 水平升高，深层神经元积累二肽重复蛋白聚 (GA)、DNA 损伤并发生核固缩，这可以通过 GSK2606414 进行药理学拯救。因此，患者特异性 iPSC 衍生的皮质类器官切片培养物是一个可重复的转化平台，用于研究临床前 ALS/FTD 机制以及新的治疗方法。