High Homocysteine-Thiolactone Leads to Reduced MENIN Protein Expression and an Impaired DNA Damage Response: Implications for Neural Tube Defects

Baoling Bai; Chunlei Wan; Zonghui Xiao; Dan Li; Lingyun Liu; Kexin Zhang; Ting Zhang; Qin Zhang

doi:10.1007/s12035-024-04033-7

High Homocysteine-Thiolactone Leads to Reduced MENIN Protein Expression and an Impaired DNA Damage Response: Implications for Neural Tube Defects

Mol Neurobiol. 2024 Feb 22. doi: 10.1007/s12035-024-04033-7. Online ahead of print.

Authors

Baoling Bai¹, Chunlei Wan², Zonghui Xiao¹, Dan Li², Lingyun Liu¹, Kexin Zhang¹, Ting Zhang³, Qin Zhang⁴

Affiliations

¹ Beijing Municipal Key Laboratory of Child Development and Nutriomics, Capital Institute of Pediatrics, Yabao Road 2, Beijing, 100020, China.
² Department of Pediatrics, Tongzhou Maternal and Child Health Care Hospital, Beijing, 101101, China.
³ Beijing Municipal Key Laboratory of Child Development and Nutriomics, Capital Institute of Pediatrics, Yabao Road 2, Beijing, 100020, China. zhangtingcv@126.com.
⁴ Beijing Municipal Key Laboratory of Child Development and Nutriomics, Capital Institute of Pediatrics, Yabao Road 2, Beijing, 100020, China. maureenzq@hotmail.com.

PMID: 38386135
DOI: 10.1007/s12035-024-04033-7

Abstract

DNA damage is associated with hyperhomocysteinemia (HHcy) and neural tube defects (NTDs). Additionally, HHcy is a risk factor for NTDs. Therefore, this study examined whether DNA damage is involved in HHcy-induced NTDs and investigated the underlying pathological mechanisms involved. Embryonic day 9 (E9) mouse neuroectoderm cells (NE4C) and homocysteine-thiolactone (HTL, active metabolite of Hcy)-induced NTD chicken embryos were studied by Western blotting, immunofluorescence. RNA interference or gene overexpression techniques were employed to investigate the impact of Menin expression changes on the DNA damage. Chromatin immunoprecipitation-quantitative polymerase chain reaction was used to investigate the epigenetic regulation of histone modifications. An increase in γH2AX (a DNA damage indicator) was detected in HTL-induced NTD chicken embryos and HTL-treated NE4C, accompanied by dysregulation of phospho-Atr-Chk1-nucleotide excision repair (NER) pathway. Further investigation, based on previous research, revealed that disruption of NER was subject to the epigenetic regulation of low-expressed Menin-H3K4me3. Overexpression of Menin or supplementation with folic acid in HTL-treated NE4C reversed the adverse effects caused by high HTL. Additionally, by overexpressing the Mars gene, we tentatively propose a mechanism whereby HTL regulates Menin expression through H3K79hcy, which subsequently influences H3K4me3 modifications, reflecting an interaction between histone modifications. Finally, in 10 human fetal NTDs with HHcy, we detected a decrease in the expression of Menin-H3K4me3 and disorder in the NER pathway, which to some extent validated our proposed mechanism. The present study demonstrated that the decreased expression of Menin in high HTL downregulated H3K4me3 modifications, further weakening the Atr-Chk1-NER pathway, resulting in the occurrence of NTDs.

Keywords: DNA damage response (DDR); Histone 3 lysine 4 trimethylation (H3K4me3); Histone 3 lysine 79 homocysteinylation (H3K79hcy); Homocysteine-thiolactone (HTL); Hyperhomocysteinemia (HHcy); Men1/Menin; Neural tube defects.

Abstract

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