Growth & Development
NAMRATA LANDGE RANA, BDS, MSD (she/her/hers)
Pediatric Dental Resident
UNIVERSITY OF ALABAMA AT BIRMINGHAM
UNIVERSITY OF ALABAMA AT BIRMINGHAM
San Diego, California, United States
Yanfang Zhao Zhao, BDS, PhD
University of Alabama at Birmingham
Greg Harber, MS
University of Alabama at Birmingham
Amjad Javed, PhD
University of Alabama at Birmingham
Janice G. Jackson, D.M.D
Program Director
University of Alabama at Birmingham
Birmingham, Alabama, United States
Ping Zhang, DDS, PhD
Professor and Director of Research
University of Alabama at Birmingham
University of Alabama at Birmingham
Birmingham, Alabama, United States
Purpose: DPY30 is a core subunit of SET1/MII complexes, the major histone H3K4 methyltransferases in mammals. It plays an important role in regulating fundamental processes including the determination of cell lineage, growth, and differentiation. Histone-based epigenetic mechanisms are involved in odontoblast differentiation and function. This study aimed to determine the role of DPY30 in the epigenetic regulation of odontoblast differentiation in vitro.
Methods: Primary human dental pulp cells (hDPCs) were used to induce odontoblast differentiation. Odontoblast differentiation and mineralization were determined by gene expression and staining with alkaline phosphate (ALP), and alizarin red (ARS). The effect of DPY30 on odontoblast differentiation and mineralization was determined by the siRNA knockdown of DPY30. The gene expression was analyzed at days 0, 3, 7, 14, and 21 of differentiation by quantitative PCR.
Results: During hDPCs differentiation, the expression of DPY30 increased from day 3 to day 21, along with a progressive increase in the expression of both early (RUNX2, COL1A, DSPP) and late (DMP1, OCN) marker genes. Differentiation of hDPCs to odontoblasts was further confirmed by ALP and ARS staining. Successful knockdown of DPY30 was achieved and confirmed by qPCR. DPY30 knockdown in hDPCs led to a marked reduction in ALP activity and odontoblast differentiation. In addition, the expression of odontogenic marker genes RUNX2, COL1A, DSPP, DMP1, and OCN was significantly reduced by DPY30 depletion.
Conclusion: DPY30 mediates H3K4 methylation and promotes odontoblast differentiation and mineralization. Our results demonstrate the critical role of DPY30 in the epigenetic regulation of odontogenic differentiation of hDPCs.