Liu Pei-Pei, Han Xiao, Li Xiao, Dai Shang-Kun, Xu Ya-Jie, Jiao Lin-Fei, Du Hong-Zhen, Zhao Li-Hua, Li Rong-Feng, Teng Zhao-Qian, Yang Yun-Gui, Liu Chang-Mei
Key Laboratory of Organ Regeneration and Reconstruction, State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, 100101, China.
University of Chinese Academy of Sciences, Beijing, 100049, China.
Adv Sci (Weinh). 2025 Jan;12(1):e2403591. doi: 10.1002/advs.202403591. Epub 2024 Nov 5.
EED (embryonic ectoderm development) is a core subunit of the polycomb repressive complex 2 (PRC2), which senses the trimethylation of histone H3 lysine 27 (H3K27). However, its biological function in cerebellar development remains unknown. Here, we show that EED deletion from neural stem cells (NSCs) or cerebellar granule cell progenitors (GCPs) leads to reduced GCPs proliferation, cell death, cerebellar hypoplasia, and motor deficits in mice. Joint profiling of transcripts and ChIP-seq analysis in cerebellar granule cells reveals that EED regulates bunches of genes involved in cerebellar development. EED ablation exhibits overactivation of a developmental repressor long non-coding RNA H19. Importantly, an obvious H3K27ac enrichment is found at Ctcf, a trans-activator of H19, and H3K27me3 enrichment at the H19 imprinting control region (ICR), suggesting that EED regulates H19 in an H3K27me3-dependent manner. Intriguingly, H19 deletion reduces EED expression and the reprogramming of EED-mediated H3K27me3 profiles, resulting in increased proliferation, differentiation, and decreased apoptosis of GCPs. Finally, molecular and genetic evidence provides that increased H19 expression is responsible for cerebellar hypoplasia and motor defects in EED mutant mice. Thus, this study demonstrates that EED, H19 forms a negative feedback loop, which plays a crucial role in cerebellar morphogenesis and controls cerebellar development.
胚胎外胚层发育蛋白(EED)是多梳抑制复合物2(PRC2)的核心亚基,可感知组蛋白H3赖氨酸27(H3K27)的三甲基化。然而,其在小脑发育中的生物学功能仍不清楚。在此,我们表明从小鼠神经干细胞(NSCs)或小脑颗粒细胞祖细胞(GCPs)中缺失EED会导致GCPs增殖减少、细胞死亡、小脑发育不全和运动缺陷。对小脑颗粒细胞中的转录本进行联合分析和ChIP-seq分析表明,EED调节一系列参与小脑发育的基因。EED缺失导致发育抑制性长链非编码RNA H19过度激活。重要的是,在H19的反式激活因子Ctcf处发现明显的H3K27ac富集,在H19印记控制区(ICR)处发现H3K27me3富集,这表明EED以H3K27me3依赖的方式调节H19。有趣的是,H19缺失会降低EED表达以及EED介导的H3K27me3图谱的重编程,导致GCPs增殖增加、分化增加和凋亡减少。最后,分子和遗传学证据表明,H19表达增加是EED突变小鼠小脑发育不全和运动缺陷的原因。因此,本研究表明EED、H19形成一个负反馈环,在小脑形态发生中起关键作用并控制小脑发育。