Pathogenic Mechanism Of Sp4 In Cleft Palate

This study aims to identify potential pathogenic transcription factors (TFs) and loci through integrated analysis of ATAC-seq and RNA-seq. A series of in vivo and in vitro experiments were conducted to elucidate the regulatory mechanisms of candidate TFs on downstream targets, clarify their pathogenic roles, and ultimately reveal their contributions to cleft palate (CP) development. These findings aim to provide theoretical foundations for the diagnosis, treatment, and intervention of CP. ATAC-seq was conducted to screen differentially accessible chromatin regions between E14.5 fetal mice with cleft palate and normal, and motif prediction was used to identify potentially key transcription factors (TFs) involved in CP. RNA-seq was performed to screen differentially expressed transcription. Cell and mouse cleft lip and palate models were used to detect the function of transcription factor, and zebrafish transgenic model was constructed to observe the cleft palate phenotype. Through the joint analysis of omics data, we systematically mined the potential risk transcription factor of CP. Taking Sp4 as the research object, we found that this transcription factor played a positive regulatory role in the process of palatal development, causing the expression of downstream protein Gdf5 to be up-regulated, and the latter changed the distribution of ion channel protein Slc39a8 in the cytoplasm. Transcription factor Sp4 up regulates the expression of downstream gene Gdf5, which changes the localization of Slc39a8 in cytoplasm, activates iron death in palatal mesenchymal cells, and ultimately affects the normal process of palatal development.