Valproic acid (VPA) is an effective and widely used anti-seizure medication but is teratogenic when used during pregnancy, affecting brain and spinal cord development for reasons that remain largely unclear. Here we designed a genetic recombinase-based SOX10 reporter system in human pluripotent stem cells that enables tracking and lineage tracing of Neural Crest cells (NCCs) in a human organoid model of the developing neural tube. We found that VPA induces extensive cellular senescence and promotes mesenchymal differentiation of human NCCs. We next show that the clinically approved drug Rapamycin inhibits senescence and restores aberrant NCC differentiation trajectory after VPA exposure in human organoids and in developing zebrafish, highlighting the therapeutic promise of this approach. Finally, we identify the pioneer factor AP1 as a key element of this process. Collectively our data reveal cellular senescence as a central driver of VPA-associated neurodevelopmental teratogenicity and identifies a new pharmacological strategy for prevention. These results exemplify the power of genetically modified human stem cell-derived organoid models for drug discovery.

丙戊酸 (VPA) 是一种有效且广泛使用的抗癫痫药物，但在怀孕期间使用会致畸，影响大脑和脊髓发育，其原因尚不清楚。在这里，我们在人类多能干细胞中设计了一种基于基因重组酶的SOX10报告系统，该系统能够在发育中的神经管的人类类器官模型中跟踪和谱系追踪神经嵴细胞 (NCC)。我们发现 VPA 诱导广泛的细胞衰老并促进人类 NCC 的间质分化。接下来，我们表明，在人类类器官和发育中的斑马鱼中暴露于 VPA 后，临床批准的药物雷帕霉素可以抑制衰老并恢复异常的 NCC 分化轨迹，这凸显了这种方法的治疗前景。最后，我们将先锋因素 AP1 确定为该过程的关键要素。总的来说，我们的数据揭示了细胞衰老是 VPA 相关神经发育致畸性的核心驱动因素，并确定了一种新的预防药理学策略。这些结果证明了转基因人类干细胞衍生的类器官模型在药物发现方面的力量。