Effect of Folate Deficiency on the Changes of Histone H3 Lysine 4 Monomethylation-Marked Enhancers and Its Molecular Exploration in Low Folate-Induced Neural Tube Defects

Qiu XIE; Jin HU; Jianting LI; Ting ZHANG

doi:10.3881/j.issn.1000-503X.16323

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Acta Academiae Medicinae Sinicae ›› 2025, Vol. 47 ›› Issue (5) : 782-791. DOI: 10.3881/j.issn.1000-503X.16323

Effect of Folate Deficiency on the Changes of Histone H3 Lysine 4 Monomethylation-Marked Enhancers and Its Molecular Exploration in Low Folate-Induced Neural Tube Defects

Qiu XIE ¹ ,
Jin HU ¹ ,
Jianting LI ² ,
Ting ZHANG ²

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Abstract

Objective To investigate the effects of folate deficiency on changes in histone H3 lysine 4 (H3K4) mono-methylation (me1)-marked enhancers and the molecular mechanism underpinning the folate deficiency-induced neural tube defects (NTD). Methods Mouse embryonic stem cells (mESCs) were cultured in the folate-free DMEM medium (folate-deficient group) and the DMEM medium containing 4 mg/L folate (normal control group),respectively.Chromatin immunoprecipitation sequencing (ChIP-seq) was performed for H3K4me1. The mouse model of folate-induced NTD was established,and transcriptome sequencing (RNA-seq) was performed for the brain tissue of fetal mice to reveal the differential expression profiles.The results were validated through real-time quantitative polymerase chain reaction (RT-qPCR).The activity of the differential peak regions of H3K4me1 was verified through the luciferase reporter assay. Results The folate content in the mESCs cultured in the folate-free medium reduced compared with that in the normal control group (P=0.008).The H3K4me1-maked enhancers in the mESCs cultured in the folate-free medium induced significant changes in intronic regions,and these changes were concentrated in metabolic and energy metabolism processes (q=9.56×10^-48,P=1.28×10^-47).The differentially expressed genes harboring H3K4me1-marked enhancers in mESCs were mainly enriched in the Wnt signaling pathway (q=0.004,P=0.004 7).ChIP-qPCR results confirmed that H3K4me1 binding decreased in the differential peak regions of the Ldlrap1 gene (P=0.008),Camta1 gene (P=0.002),and Apc2 gene (P=0.012).The H3K4 demethylase inhibitor T-448 effectively reversed the H3K4me1 binding in the differential peak regions of the aforementioned genes (P=0.01).The results of RNA-seq for the brain tissue of NTD fetal mice showed significant enrichment of the differentially expressed genes in the Wnt signaling pathway (P=1.52×10^-5).The enrichment of differential peak regions of H3K4me1-marked enhancers in Apc2,Ldlrap1,and Camta1 genes in the brain tissue also showed significant changes.The differential peak region in Apc2 exhibited transcription factor activity (P=0.020). Conclusion Folate deficiency may affect changes in H3K4me1-marked enhancers to participate in the regulation of neural tube closure genes,thereby inducing the occurrence of NTD.

Key words

histone / folate / embryonic stem cells / neural tube defects / chromatin immunoprecipitation

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Qiu XIE , Jin HU , Jianting LI , et al. Effect of Folate Deficiency on the Changes of Histone H3 Lysine 4 Monomethylation-Marked Enhancers and Its Molecular Exploration in Low Folate-Induced Neural Tube Defects[J]. Acta Academiae Medicinae Sinicae. 2025, 47(5): 782-791 https://doi.org/10.3881/j.issn.1000-503X.16323

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