The beneficial effect of global O-GlcNAcylation on odontogenic differentiation of human dental pulp cells via mTORC1 pathway

Abstract

ObjectiveTo investigate whether and how global O-linked N-Acetylglucosamine modification (O-GlcNAcylation), a prevalent nutrient-sensitive post-translation modification, regulates odontogenic differentiation and mineralization in human dental pulp cells (hDPCs). DesignFirst, immunostaining assays on sections of dental pulp tissue were performed to detect the distributions of O-GlcNAcylation and its exclusive enzyme set O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA). Then global O-GlcNAcylation was determined by anti O-linked N-Acetylglucosamine (RL2) Western blot during odontogenesis of hDPCs. Further, inhibition or knockdown of OGT and OGA were achieved by specific inhibitors or siRNA in vitro, respectively. The odonto-induction effect of O-GlcNAcylation ex vivo was investigated by a subcutaneous transplantation experiment. Moreover, the O-GlcNAc modification of RAPTOR was confirmed by immunoprecipitation. Odontogenic differentiation assays also investigated the indispensable role of RAPTOR during enhanced global O-GlcNAcylation. ResultsThe signals of O-GlcNAc became more enriched in the odontoblasts compared to pulp fibroblasts. During odontogenesis of hDPCs, global O-GlcNAcylation was significantly increased. An increase or decrease of O-GlcNAcylation significantly boosted or blunted odontogenic differentiation, respectively. The fluctuation of O-GlcNAcylation continuously impacted the downstream targets of mTORC1. Consistently, RAPTOR was modified by O-GlcNAcylation, which was necessary for inducing odontogenesis. ConclusionsGlobal O-GlcNAcylation participated in and affected the odontogenic differentiation of hDPCs, which was mediated by the mTORC1 pathway. Thus, targeting O-GlcNAcylation might be a potential therapeutic intervention for pulp repair and regeneration.