title: KDM6B epigenetically regulates odontogenic differentiation of dental mesenchymal stem cells.
abstract: Mesenchymal stem cells (MSCs) have been identified and isolated from dental tissues, including stem cells from apical papilla, which demonstrated the ability to differentiate into dentin-forming odontoblasts. The histone demethylase KDM6B (also known as JMJD3) was shown to play a key role in promoting osteogenic commitment by removing epigenetic marks H3K27me3 from the promoters of osteogenic genes. Whether KDM6B is involved in odontogenic differentiation of dental MSCs, however, is not known. Here, we explored the role of KDM6B in dental MSC fate determination into the odontogenic lineage. Using shRNA-expressing lentivirus, we performed KDM6B knockdown in dental MSCs and observed that KDM6B depletion leads to a significant reduction in alkaline phosphate (ALP) activity and in formation of mineralized nodules assessed by Alizarin Red staining. Additionally, mRNA expression of odontogenic marker gene SP7 (osterix, OSX), as well as extracellular matrix genes BGLAP (osteoclacin, OCN) and SPP1 (osteopontin, OPN), was suppressed by KDM6B depletion. When KDM6B was overexpressed in KDM6B-knockdown MSCs, odontogenic differentiation was restored, further confirming the facilitating role of KDM6B in odontogenic commitment. Mechanistically, KDM6B was recruited to bone morphogenic protein 2 (BMP2) promoters and the subsequent removal of silencing H3K27me3 marks led to the activation of this odontogenic master transcription gene. Taken together, our results demonstrated the critical role of a histone demethylase in the epigenetic regulation of odontogenic differentiation of dental MSCs. KDM6B may present as a potential therapeutic target in the regeneration of tooth structures and the repair of craniofacial defects.
authors.affiliation: 1] Department of Stomatology, Chinese People's Liberation Army General Hospital, Beijing, China [2] Division of Oral Biology and Medicine, School of Dentistry, University of California, Los Angeles, USA.
authors.forename: Juan
authors.initials: J
authors.lastname: Xu
authors.forename: Bo
authors.initials: B
authors.lastname: Yu
authors.forename: Christine
authors.initials: C
authors.lastname: Hong
authors.forename: Cun-Yu
authors.initials: CY
authors.lastname: Wang
mesh_terms: D000469:Alkaline Phosphatase / Q000032:analysis; D055396:Bone Morphogenetic Protein 2 / Q000235:genetics; D055415:Bone Morphogenetic Protein 4 / Q000235:genetics; D002113:Calcification, Physiologic / Q000235:genetics; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000235:genetics; D019070:Cell Lineage; D003771:Dental Papilla / Q000166:cytology*; D044127:Epigenesis, Genetic / Q000235:genetics*; D055785:Gene Knockdown Techniques; D018398:Homeodomain Proteins / Q000235:genetics; D006801:Humans; D056484:Jumonji Domain-Containing Histone Demethylases / Q000235:genetics*; D059630:Mesenchymal Stem Cells / Q000502:physiology*; D009804:Odontoblasts / Q000502:physiology; D009805:Odontogenesis / Q000235:genetics*; D015675:Osteocalcin / Q000032:analysis; D053495:Osteopontin / Q000032:analysis; D011401:Promoter Regions, Genetic / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics; D000076182:Sp7 Transcription Factor; D014157:Transcription Factors / Q000032:analysis / Q000235:genetics; D015533:Transcriptional Activation / Q000235:genetics
chemical_list: C525718:BMP2 protein, human; C525809:BMP4 protein, human; D055396:Bone Morphogenetic Protein 2; D055415:Bone Morphogenetic Protein 4; C088334:Distal-less homeobox proteins; D018398:Homeodomain Proteins; D034741:RNA, Small Interfering; C506895:SPP1 protein, human; D000076182:Sp7 Transcription Factor; C478163:SP7 protein, human; D014157:Transcription Factors; D015675:Osteocalcin; D053495:Osteopontin; D056484:Jumonji Domain-Containing Histone Demethylases; C524220:KDM6B protein, human; D000469:Alkaline Phosphatase