慢性钴暴露诱导人大脑类器官神经毒性的ceRNA调控机制

doi:10.3969/j.issn.1004-616x.2025.06.004

癌变·畸变·突变 ›› 2025, Vol. 37 ›› Issue (6): 445-451,457.doi: 10.3969/j.issn.1004-616x.2025.06.004

• 论著 • 上一篇

慢性钴暴露诱导人大脑类器官神经毒性的ceRNA调控机制

郭新华, 黄琰, 陈琪琪, 鲁诗雅, 曾品利, 王志秋, 李灏, 卜迁

四川大学华西公共卫生学院/华西第四医院卫生毒理与病理学系, 四川成都 610041

收稿日期:2025-05-15 修回日期:2025-07-08 发布日期:2025-12-06
通讯作者: 卜迁，E-mail：buqian7978@scu.edu.cn
作者简介:郭新华，E-mail：guoxinhua2022@163.com。
基金资助:
国家自然科学基金原创探索计划项目(T2350007)

ceRNA regulatory mechanisms in neurotoxicity induced by chronic cobalt exposure in cerebral organoids

GUO Xinhua, HUANG Yan, CHEN Qiqi, LU Shiya, ZENG Pinli, WANG Zhiqiu, LI Hao, BU Qian

Department of Health Toxicology and Pathology, West China School of Public Health, Sichuan University/West China Fourth Hospital, Chengdu 610041, Sichuan, China

Received:2025-05-15 Revised:2025-07-08 Published:2025-12-06

摘要/Abstract

摘要： 目的：基于全转录组测序和共表达分析，探究慢性钴暴露条件下非编码RNA(ncRNA)表达谱的改变以及竞争性内源RNA(ceRNA)网络在钴诱导神经毒性中的作用。方法：将培养成熟的人大脑类器官分别于含0、20 mol/L CoCl₂的培养基中培养28 d，采用NovaSeq X Plus平台对人大脑类器官进行全转录组测序，以[log₂FC]≥1，P_校正后<0.05为筛选阈值，识别对照组与钴暴露组间差异基因和差异ncRNA，并进行基因本体(GO)富集分析。根据Spearman相关性系数筛选具有潜在相互作用的ceRNA构建ceRNA网络。并对网络中的部分RNA进行反转录实时荧光定量PCR(RT-qPCR)验证。结果：全转录组测序共筛选出2 968个差异基因、 2 832个差异lncRNA、82个差异miRNA和35个差异circRNA。GO分析表明差异基因和差异ncRNA靶基因均与代谢物和能量产生相关，主要包括ATP生物合成、线粒体呼吸链复合物、电子传递链等。基于iPSC衍生神经元的进一步验证表明，钴暴露可导致细胞内ATP合成减少。最后构建特定于能量代谢基因集的ceRNA调控网络，共包括11个mRNA、10个miRNA、30个lncRNA和2个circRNA。RT-qPCR验证表明丙酮酸激酶和miRNA 17_9658的表达趋势相反，且与测序结果一致。结论：慢性钴暴露可能通过影响代谢通路诱导神经毒性，而ceRNA调控网络可能在重金属钴的神经毒性机制中发挥重要作用。

关键词: 钴, 人大脑类器官, 全转录组测序, ceRNA网络

Abstract: OBJECTIVE：Based on whole transcriptome sequencing and co-expression analysis，this study aimed to explore changes in non-coding RNA (ncRNA) expression profiles under chronic cobalt exposure to cerebral organoids and to investigate the role of competitive endogenous RNA (ceRNA) network in cobalt-induced neurotoxicity. METHODS：Mature cerebral organoids were continuously exposed to 0 and 20 μmol/L CoCl₂ for 28 days. Whole transcriptome sequencing was performed using the NovaSeq X Plus platform. Differential genes and differential ncRNAs between the control group and cobalt exposure group were identified with the screening thresholds of [log₂FC]≥1 and P_adjust<0.05，followed by gene ontology (GO) enrichment analysis. CeRNAs with potential interactions were screened according to Spearman correlation coefficient to construct a ceRNA network. Some RNAs in the network were verified by real-time quantitative PCR (RT-qPCR). RESULTS：A total of 2 968 differential genes，2 832 differential lncRNAs，82 differential miRNAs，and 35 differential circRNAs were identified by whole transcriptome sequencing. GO analysis showed that differential genes and target genes of differential ncRNAs were associated with metabolites and energy production，mainly including ATP biosynthesis，mitochondrial respiratory chain complex，and electron transport chain. Further verification using iPSC-derived neurons indicated that cobalt exposure could reduce intracellular ATP synthesis. Finally，a ceRNA regulatory network specific to the energy metabolism gene set was constructed，including 11 mRNAs，10 miRNAs，30 lncRNAs，and 2 circRNAs. RT-qPCR verification showed that the expression trends of PKM and miRNA 17_9658 were opposite，which was consistent with the sequencing results. CONCLUSION：Chronic cobalt exposure may induce neurotoxicity by affecting metabolic pathways，and the ceRNA regulatory network may play an important role in the neurotoxic mechanism of heavy metal cobalt.

Key words: cobalt, cerebral organoids, whole transcriptome sequencing, ceRNA network

中图分类号:

R3
R994.6

郭新华, 黄琰, 陈琪琪, 鲁诗雅, 曾品利, 王志秋, 李灏, 卜迁. 慢性钴暴露诱导人大脑类器官神经毒性的ceRNA调控机制[J]. 癌变·畸变·突变, 2025, 37(6): 445-451,457.

GUO Xinhua, HUANG Yan, CHEN Qiqi, LU Shiya, ZENG Pinli, WANG Zhiqiu, LI Hao, BU Qian. ceRNA regulatory mechanisms in neurotoxicity induced by chronic cobalt exposure in cerebral organoids[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2025, 37(6): 445-451,457.

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慢性钴暴露诱导人大脑类器官神经毒性的ceRNA调控机制

ceRNA regulatory mechanisms in neurotoxicity induced by chronic cobalt exposure in cerebral organoids

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