Xu Jing, Mao Xiao, Liu Zhen, Jiang Na, Wong Xin E, Liu Deng, Wang Yuan, Zhan Huaizhe, Liu Shiyi, Yu Jiayao, Yuan Ruiying, Bai Qingran, Bai Xianshu, Huang Wenhui, Xie Ruoxiao, Krenn Veronica, Kirchhoff Frank, Wang Hua, Guo Zhenming, Bian Shan
Institute for Regenerative Medicine, Medical Innovation Center and State Key Laboratory of Cardiovascular Diseases, Shanghai East Hospital, National Stem Cell Translational Resource Center & Ministry of Education Stem Cell Resource Center, Frontier Science Center for Stem Cell Research, School of Life Sciences and Technology, Tongji University, Shanghai, China.
National Health Commission Key Laboratory of Birth Defect Research and Prevention, Hunan Provincial Maternal and Child Health Care Hospital, University of South China, Changsha, China.
EMBO Mol Med. 2025 Sep 8. doi: 10.1038/s44321-025-00302-7.
Primary microcephaly, a rare congenital condition characterized by reduced brain size, occurs due to impaired neurogenesis during brain development. Through whole-exome sequencing, we identified compound heterozygous loss-of-function mutations in CENTRIN 3 (CETN3) in a 5-year-old patient with primary microcephaly. As CETN3 has not been previously linked to microcephaly, we investigated its potential function in neurodevelopment in human pluripotent stem cell-derived cerebral organoids. We showed that CETN3-knockout (KO) organoids successfully recapitulated the microcephaly phenotype of reduced size compared to the control organoids. Through transcriptomic, histological, and protein analyses, we found that CETN3 deficiency directly interferes with neuronal differentiation and reduces proliferative capacity in neural stem/progenitor cells by impairing centrosome assembly required in cell cycle progression, consequently activating apoptosis. Furthermore, our data uncovered previously undocumented indirect effects of CETN3 through interaction with RNA splicing machinery involved in brain development. These findings expand the scope of known regulatory mechanisms of CETN3 in brain development and its etiological roles in human brain malformation.
原发性小头畸形是一种罕见的先天性疾病，其特征是脑容量减小，是由于大脑发育过程中神经发生受损所致。通过全外显子组测序，我们在一名5岁的原发性小头畸形患者中鉴定出中心粒蛋白3（CETN3）的复合杂合功能丧失突变。由于此前CETN3尚未与小头畸形相关联，我们在人多能干细胞衍生的类脑器官中研究了其在神经发育中的潜在功能。我们发现，与对照类脑器官相比，CETN3基因敲除（KO）类脑器官成功再现了小头畸形的体积减小表型。通过转录组学、组织学和蛋白质分析，我们发现CETN3缺乏直接干扰神经元分化，并通过损害细胞周期进程所需的中心体组装来降低神经干/祖细胞的增殖能力，从而激活细胞凋亡。此外，我们的数据揭示了CETN3通过与参与脑发育的RNA剪接机制相互作用产生的此前未被记录的间接影响。这些发现扩展了CETN3在脑发育中的已知调控机制范围及其在人类脑畸形中的病因学作用。
