尽管神经外科手术已经得到发展，但从头部外伤性脑损伤（TBI）中恢复功能的治疗选择很少。神经干/祖细胞（NS/PCs）可能对神经恢复产生长期影响。虽然诱导多能干细胞（iPSCs）可以克服在临床应用中用于人类胚胎或胎儿来源的组织的道德和实际问题，但iPSCs源从恶性肿瘤中仍然存在障碍。本文提出了一种使用表达酵母胞嘧啶脱氨酶-尿嘧啶磷酸核糖转移酶（yCD-UPRT）酶-药物基因的iPSCs治疗TBI的新策略。由人类iPSCs衍生的NS/PCs经过CRISPR/Cas9介导的基因组编辑后，表达yCD-UPRT的转基因表达稳定而高。体内生物发光成像和组织病理学分析表明，NS/PCs在TBI小鼠模型的损伤皮质周围浓集。在亚急性期期间，相对于对照组，移植基因组编辑的iPSCs衍生的NS/PCs的治疗组的射束步行测试和加速转子测试表现显著改善。埃文斯蓝外漏可视化的损伤区在治疗组中比对照组小，表明防止了继发性脑损伤。在慢性期间，治疗组的脑萎缩和脑室扩大明显减少。此外，在5-氟胞嘧啶（5-FC）给药后，由5-FC转化而来的5-氟尿嘧啶有选择地消除不成熟的NS/PCs，同时保留邻近的神经结构。表达yCD-UPRT的NS/PCs可以应用于安全的再生医学，不必担心肿瘤发生。 ©2023作者（们）。由牛津大学出版社出版。版权所有。请发送电子邮件至journals.permissions@oup.com获取授权。

Despite developing neurosurgical procedures, few treatment options have achieved functional recovery from traumatic brain injury (TBI). Neural stem/progenitor cells (NS/PCs) may produce a long-term effect on neurological recovery. Although induced pluripotent stem cells (iPSCs) can overcome ethical and practical issues of human embryonic or fetal-derived tissues in clinical applications, the tumorigenicity of iPSC-derived populations remains an obstacle to their safe use in regenerative medicine. We herein established a novel treatment strategy for TBI using iPSCs expressing the enzyme-prodrug gene yeast cytosine deaminase-uracil phosphoribosyl transferase (yCD-UPRT). NS/PCs derived from human iPSCs displayed stable and high transgene expression of yCD-UPRT following CRISPR/Cas9-mediated genome editing. In vivo bioluminescent imaging and histopathological analysis demonstrated that NS/PCs concentrated around the damaged cortex of the TBI mouse model. During the subacute phase, performances in both beam walking test and accelerating rotarod test were significantly improved in the treatment group transplanted with genome-edited iPSC-derived NS/PCs compared with the control group. The injury area visualized by extravasation of Evans blue was smaller in the treatment group compared with the control group, suggesting the prevention of secondary brain injury. During the chronic phase, cerebral atrophy and ventricle enlargement were significantly less evident in the treatment group. Furthermore, after 5-fluorocytosine (5-FC) administration, 5-fluorouracil converted from 5-FC selectively eliminated undifferentiated NS/PCs while preserving the adjacent neuronal structures. NS/PCs expressing yCD-UPRT can be applied for safe regenerative medicine without the concern for tumorigenesis.© The Author(s) 2023. Published by Oxford University Press. All rights reserved. For permissions, please email: journals.permissions@oup.com.