DLX3 epigenetically regulates odontoblastic differentiation of hDPCs through H19/miR-675 axis

Abstract

ObjectivesA novel mutation (c.533 A > G; Q178R) in DLX3 gene is responsible for Tricho-Dento-Osseous (TDO) syndrome. As one of features of TDO syndrome is dentin hypoplasia, we explored the mechanism regarding dentin defects in TDO syndrome. DesignhDPCs were obtained from the healthy premolars, stably expressing hDPCs were generated using recombinant lentiviruses. Quantitative methylation analysis, DNMT3B activity, CHIP, and evaluation of odonto-differentiation ability of hDPCs assays were performed. ResultsNovel mutant DLX3 (MU-DLX3) significantly inhibited the expression of long non-coding RNA H19 and resulted in hyper-methylation of H19 in MU group, rescue studies showed that up-regulation the expression of H19 and demethylation of H19 in MU group were able to rescue the effect of MU-DLX3. Subsequently, miR-675, encoded by H19, was also able to rescue the above effects of MU-DLX3. Thus, we proposed that MU-DLX3 regulated odontoblastic differentiation of hDPCs through H19/miR-675 axis. Through CHIP and DNMT3B activity assays disclosed the underlying mechanism by which MU-DLX3 altered H19 expression and methylation status in MU group by increasing H3K9me3 enrichment and DNMT3B activity. ConclusionsOur new findings, for the first time, suggest that MU-DLX3 significantly inhibits hDPCs differentiation via H19/miR-675 axis and provides a new mechanism insight into how MU-DLX3 epigenetically alters H19 methylation status and expression contributes to dentin hypoplasia in TDO syndrome.