Gcm1 Orchestrates Lef1 Expression in Folate Deficiency-Induced Neural Tube Defects

Neural tube defects (NTDs) are severe congenital anomalies with limited therapeutic options. Increased expression of the transcription factor glial cell missing 1 (Gcm1) has been linked to NTDs, yet the mechanisms underlying this association remain elusive. Herein, folate deficiency upregulated Gcm1 expression through H4 acetylation (H4ac) enrichment in the promoter. ChIP-qPCR and Co-IP experiments revealed that folate deficiency enhanced H4 acetylation at the Gcm1 promoter via CREB-binding protein (CBP), subsequently resulting in increased Gcm1 transcription. Furthermore, Gcm1-ChIP-seq revealed lymphoid enhancer binding factor 1 (Lef1) as a potential downstream target of Gcm1 in mESCs without folate supplementation and the likely activator of the Wnt/β-catenin pathway for aberrant neurodevelopment. Immunohistochemistry and immunofluorescence analyses revealed significantly higher Gcm1 and Lef1 expression in a low-folate NTD mouse model than in controls. Finally, NanoString analysis of human low-folate NTD samples confirmed a positive correlation between upregulated Gcm1 expression and Lef1 expression. Collectively, these findings indicate that the CBP-dependent Gcm1 regulation of Lef1 expression is crucial in folate-deficient NTDs, revealing the potential mechanism by which NTDs are induced by low folate concentrations and providing a promising target for therapeutic intervention.