Chen Liping, Li Ying, Yu Yan, Cai Mingze, Li Hao, Huang Minghe, Yang Guochao, Guo Jiageng, Wang Huailin, Song Zhihong, Shen Wei, Jiang Huihui, Wu Haitao
Department of Neurobiology, Beijing Institute of Basic Medical Sciences, Beijing, 100850, China.
Key Laboratory of Neuroregeneration, Co-innovation Center of Neuroregeneration, Nantong University, Nantong, Jiangsu Province, 226019, China.
Adv Sci (Weinh). 2025 Aug;12(32):e04200. doi: 10.1002/advs.202504200. Epub 2025 May 31.
Elongation Factor Tu GTP-Binding Domain Containing 2 (EFTUD2), a core spliceosomal GTPase associated with Mandibulofacial Dysostosis with Microcephaly (MFDM), plays a mechanistically undefined role in cerebral development. To investigate its pathophysiological contributions, murine models are generated through conditional Eftud2 ablation and in utero electroporation of human pathogenic EFTUD2 variants into cortical neural stem cells (NSCs). Embryonic NSC-specific Eftud2 knockout resulted in cortical disorganization and microcephaly, while pathogenic variants led to significant neuronal loss. Integrative transcriptomic and immunofluorescence analyses revealed that Eftud2 deficiency triggers apoptotic pathways, contributing to cortical malformations. Mechanistic studies using RNA co-immunoprecipitation and full-length transcriptome sequencing demonstrated that Eftud2 directly interacts with Caspase3 and Aifm1 transcripts, regulating their alternative splicing to generate pro-apoptotic isoforms. Splicing assays functionally validated this regulatory mechanism, showing its role in activating cell death pathways and disrupting neurodevelopmental homeostasis. These findings elucidate EFTUD2's critical role in maintaining apoptotic balance during corticogenesis and identify defective splicing regulation as the molecular basis of MFDM. This study provides insights for advancing diagnostic frameworks and therapeutic strategies for neurodevelopmental disorders.
含延伸因子Tu GTP结合结构域2（EFTUD2）是一种与小头畸形下颌面骨发育不全（MFDM）相关的核心剪接体GTP酶，在大脑发育中发挥着机制不明的作用。为了研究其病理生理学贡献，通过条件性Eftud2基因敲除以及将人类致病性EFTUD2变体在子宫内电穿孔导入皮质神经干细胞（NSC）来构建小鼠模型。胚胎期NSC特异性Eftud2基因敲除导致皮质结构紊乱和小头畸形，而致病性变体则导致显著的神经元丢失。综合转录组学和免疫荧光分析表明，Eftud2缺乏会触发凋亡途径，导致皮质畸形。使用RNA免疫共沉淀和全长转录组测序的机制研究表明，Eftud2直接与Caspase3和Aifm1转录本相互作用，调节它们的可变剪接以产生促凋亡异构体。剪接分析从功能上验证了这一调节机制，表明其在激活细胞死亡途径和破坏神经发育稳态中的作用。这些发现阐明了EFTUD2在皮质发生过程中维持凋亡平衡的关键作用，并确定剪接调控缺陷是MFDM的分子基础。本研究为推进神经发育障碍的诊断框架和治疗策略提供了见解。
