Aggregated α-synuclein (α-SYN) proteins, encoded by the SNCA gene, are hallmarks of Lewy body disease (LBD), affecting multiple brain regions. However, the specific mechanisms underlying α-SYN pathology in cortical neurons, crucial for LBD-associated dementia, remain unclear. Here, we recapitulated α-SYN pathologies in human induced pluripotent stem cells (iPSCs)–derived cortical organoids generated from patients with LBD with SNCA gene triplication. Single-cell RNA sequencing, combined with functional and molecular validation, identified synaptic and mitochondrial dysfunction in excitatory neurons exhibiting high expression of the SNCA gene, aligning with observations in the cortex of autopsy-confirmed LBD human brains. Furthermore, we screened 1280 Food and Drug Administration–approved drugs and identified four candidates (entacapone, tolcapone, phenazopyridine hydrochloride, and zalcitabine) that inhibited α-SYN seeding activity in real-time quaking-induced conversion assays with human brains, reduced α-SYN aggregation, and alleviated mitochondrial dysfunction in SNCA triplication organoids and excitatory neurons. Our findings establish human cortical LBD models and suggest potential therapeutic drugs targeting α-SYN aggregation for LBD.

中文翻译：

---

使用 SNCA 三倍体 iPSC 衍生的皮质类器官对路易体病进行建模并鉴定治疗药物


由 SNCA 基因编码的聚集 α-突触核蛋白 （α-SYN） 蛋白是路易体病 （LBD） 的标志，影响多个大脑区域。然而，皮质神经元中 α-SYN 病理学的具体机制仍不清楚，这对 LBD 相关痴呆至关重要。在这里，我们概括了 SNCA 基因三倍的 LBD 患者产生的人诱导多能干细胞 （iPSC） 衍生的皮质类器官中的 α-SYN 病理。单细胞 RNA 测序与功能和分子验证相结合，在表现出 SNCA 基因高表达的兴奋性神经元中鉴定出突触和线粒体功能障碍，与尸检证实的 LBD 人脑皮层中的观察结果一致。此外，我们筛选了 1280 种美国食品药品监督管理局批准的药物，并确定了四种候选药物 （恩他卡朋、托卡朋、盐酸非那吡啶和扎西他滨），它们在人脑实时震动诱导的转换测定中抑制 α-SYN 接种活性，减少 α-SYN 聚集，并减轻 SNCA 三重复类器官和兴奋性神经元中的线粒体功能障碍。我们的研究结果建立了人类皮层 LBD 模型，并提出了针对 LBD 的 α-SYN 聚集的潜在治疗药物。