Otx2基因通过调节H3K27me3对Wnt通路的影响

doi:10.19586/j.2095-2341.2024.0178

生物技术进展 ›› 2025, Vol. 15 ›› Issue (3): 526-534.DOI: 10.19586/j.2095-2341.2024.0178

Otx2基因通过调节H3K27me3对Wnt通路的影响

朱媛智¹^,²(), 裴培², 刘帆², 何学佳¹, 王怡³, 王珊¹^,²()

^1.北京大学首都儿科研究所教学医院，北京 100020
^2.首都儿科研究所附属儿童医院，北京 100020
^3.首都医科大学附属北京儿童医院，北京 100045

收稿日期:2024-11-12 接受日期:2024-12-27 出版日期:2025-05-25 发布日期:2025-07-01
通讯作者: 王珊
作者简介:朱媛智 E-mail: zhuyuanzhizyz@139.com；
基金资助:
国家自然科学基金面上项目(82071690);北京市自然科学基金面上项目(7242014);北京市财政局第四批试点项目(BMR2021-3);北京市医院管理中心第四批“登峰”计划(DFL20221102)

The Otx2 Gene Regulates the Effect of H3K27me3 on the Wnt Pathway

Yuanzhi ZHU¹^,²(), Pei PEI², Fan LIU², Xuejia HE¹, Yi WANG³, Shan WANG¹^,²()

^1.Capital Institute of Pediatrics-Peking University Teaching Hospital，Beijing 100020，China
^2.Children's Hospital Capital Institute of Pediatrics，Beijing 100020，China
^3.Beijing Children's Hospital，Capital Medical University，Beijing 100045，China

Received:2024-11-12 Accepted:2024-12-27 Online:2025-05-25 Published:2025-07-01
Contact: Shan WANG

摘要/Abstract

摘要：

Otx2与Wnt通路都参与神经系统的发育过程，H3K27me3富集状态的改变也会影响神经系统的发育。为探究Otx2是否可以通过改变组蛋白H3K27me3状态调控Wnt通路，采用CRISPR/Cas9系统构建稳定敲除Otx2基因的C17.2小鼠神经干细胞株，利用ChIP-Seq技术分析H3K27me3修饰特异性结合的DNA片段，通过qPCR技术检测Otx2基因敲除后Wnt基因的表达改变。经GO富集分析与KEGG富集分析结果显示，Otx2敲除组与对照组的ChIP-Seq差异富集峰所对应的差异基因主要集中于神经系统发育与功能相关的通路。Wnt通路中的Wnt5a、Wnt2、Wnt16基因位点发生H3K27me3富集情况改变。qPCR结果显示，相比于对照组，Otx2基因敲除组的Wnt5a、Wnt2、Wnt5b基因的表达量下降。结果表明，Otx2基因通过调节组蛋白H3K27me3的富集位点影响Wnt通路，可为神经系统发育异常相关研究提供理论参考。

关键词: Otx2基因, H3K27me3, Wnt通路

Abstract:

Both Otx2 and the Wnt pathway are involved in nervous system development， and changes in H3K27me3 enrichment status are known to influence this process. This study investigated whether Otx2 regulates the Wnt pathway by modulating histone H3K27me3 modification. A stable C17.2 mouse neural stem cell line with Otx2 knockout was generated using CRISPR/Cas9. ChIP-Seq analysis identified DNA fragments bound by H3K27me3， and qPCR was used to assess Wnt-related gene expression changes after Otx2 knockout. GO and KEGG enrichment analyses revealed that differential genes associated with ChIP-Seq peaks between Otx2-knockout and control cells were predominantly enriched in pathways linked to nervous system development and function. Further analysis showed H3K27me3 enrichment at loci of Wnt5a， Wnt2， and Wnt16 genes in the Wnt pathway. Notably， Wnt5a， Wnt2， and Wnt5b expression levels were significantly reduced in Otx2-knockout cells compared to controls. These findings suggested that Otx2 regulates the Wnt pathway by altering H3K27me3 enrichment sites， providing mechanistic insights into neurological developmental abnormalities.

Key words: Otx2, H3K27me3, Wnt pathway

中图分类号:

Q189

朱媛智, 裴培, 刘帆, 何学佳, 王怡, 王珊. Otx2基因通过调节H3K27me3对Wnt通路的影响[J]. 生物技术进展, 2025, 15(3): 526-534.

Yuanzhi ZHU, Pei PEI, Fan LIU, Xuejia HE, Yi WANG, Shan WANG. The Otx2 Gene Regulates the Effect of H3K27me3 on the Wnt Pathway[J]. Current Biotechnology, 2025, 15(3): 526-534.

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Otx2基因通过调节H3K27me3对Wnt通路的影响

The Otx2 Gene Regulates the Effect of H3K27me3 on the Wnt Pathway

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