Zbtb20 Regulates Tooth Development By Modulating Eda Pathway

Transcription factor ZBTB20 is essential for the development of multiple organ systems. Mutations in the ZBTB20 gene cause Primrose Syndrome in humans, characterized by multiple developmental defects, including in teeth. This study aims to elucidate the role of ZBTB20 in tooth development. ZBTB20 expression was assessed by immunofluorescence staining, and epithelium-specific Zbtb20 gene knockout mice were generated. The dental phenotype was analyzed using Micro-CT and histological analysis. Cell proliferation and migration were evaluated using BrdU and EdU incorporation assays. Transcriptional profile was analyzed using RNA-seq and differentially expressed genes were validated by qRT-PCR, in situ hybridization, and immunofluorescence. CUT&Tag assay was employed to identify genome-wide DNA binding sites of ZBTB20. ZBTB20 protein was first detected in labial cervical loop, the epithelial stem cell niche of the incisor, at E14.5, and its expression persisted into postnatal and adult stages. Zbtb20 deletion resulted in defects in incisor development, including enamel hypoplasia characterized by a chalky appearance, reduced enamel volume and delayed mineralization revealed by Micro-CT analysis . Moreover, Zbtb20-deficient mice exhibited increased proliferation rate and delayed differentiation in incisor epithelial cells. Mechanistically, loss of ZBTB20 resulted in upregulation of EDA expression and its downstream NF-κB pathway as revealed by RNA-seq, confirmed by qRT-PCR and in situ hybridization. Furthermore, ZBTB20 occupied the promoter region of EDA gene in vivo demonstrated by CUT&Tag assays . ZBTB20 regulates the development of dental epithelial stem cells via repressing EDA/NF-κB signaling pathway.