Alcohol-induced suppression of KDM6B dysregulates the mineralization potential in dental pulp stem cells.
Epigenetic changes, such as alteration of DNA methylation patterns, have been proposed as a molecular mechanism underlying the effect of alcohol on the maintenance of adult stem cells. We have performed genome-wide gene expression microarray and DNA methylome analysis to identify molecular alterations via DNA methylation changes associated with exposure of human dental pulp stem cells (DPSCs) to ethanol (EtOH). By combined analysis of the gene expression and DNA methylation, we have found a significant number of genes that are potentially regulated by EtOH-induced DNA methylation. As a focused approach, we have also performed a pathway-focused RT-PCR array analysis to examine potential molecular effects of EtOH on genes involved in epigenetic chromatin modification enzymes, fibroblastic markers, and stress and toxicity pathways in DPSCs. We have identified and verified that lysine specific demethylase 6B (KDM6B) was significantly dysregulated in DPSCs upon EtOH exposure. EtOH treatment during odontogenic/osteogenic differentiation of DPSCs suppressed the induction of KDM6B with alterations in the expression of differentiation markers. Knockdown of KDM6B resulted in a marked decrease in mineralization from implanted DPSCs in vivo. Furthermore, an ectopic expression of KDM6B in EtOH-treated DPSCs restored the expression of differentiation-related genes. Our study has demonstrated that EtOH-induced inhibition of KDM6B plays a role in the dysregulation of odontogenic/osteogenic differentiation in the DPSC model. This suggests a potential molecular mechanism for cellular insults of heavy alcohol consumption that can lead to decreased mineral deposition potentially associated with abnormalities in dental development and also osteopenia/osteoporosis, hallmark features of fetal alcohol spectrum disorders.
Laboratory of Stem Cell & Cancer Epigenetic Research, School of Dentistry, UCLA, 10833 Le Conte Avenue, 73-041 CHS, Los Angeles, CA 90095, USA.
Michael
M
Hoang
Laboratory of Stem Cell & Cancer Epigenetic Research, School of Dentistry, UCLA, 10833 Le Conte Avenue, 73-041 CHS, Los Angeles, CA 90095, USA.
Jeffrey J
JJ
Kim
Laboratory of Stem Cell & Cancer Epigenetic Research, School of Dentistry, UCLA, 10833 Le Conte Avenue, 73-041 CHS, Los Angeles, CA 90095, USA.
Yiyoung
Y
Kim
Laboratory of Stem Cell & Cancer Epigenetic Research, School of Dentistry, UCLA, 10833 Le Conte Avenue, 73-041 CHS, Los Angeles, CA 90095, USA.
Elizabeth
E
Tong
Department of Biology, California State University at Northridge, 18111 Nordhoff Street, Northridge, CA 91330, USA.
Benjamin
B
Trammell
Department of Anatomy and Cell Biology, University of Pennsylvania School of Dental Medicine, 240 South 40th Street, Philadelphia, PA 19104, USA.
Yao
Y
Liu
Department of Anatomy and Cell Biology, University of Pennsylvania School of Dental Medicine, 240 South 40th Street, Philadelphia, PA 19104, USA.
Songtao
S
Shi
Section of Orthodontics, School of Dentistry, UCLA, 10833 Le Conte Avenue, 63-041 CHS, Los Angeles, CA 90095, USA.
Chang-Ryul
CR
Lee
Section of Orthodontics, School of Dentistry, UCLA, 10833 Le Conte Avenue, 63-041 CHS, Los Angeles, CA 90095, USA.
Christine
C
Hong
Laboratory of Molecular Signaling, Division of Oral Biology and Medicine, School of Dentistry, UCLA, 10833 Le Conte Avenue, 33-030 CHS, Los Angeles, CA 90095, USA; UCLA Broad Stem Cell Research Center, Box 957357, Los Angeles, CA 90095, USA.
Cun-Yu
CY
Wang
Laboratory of Stem Cell & Cancer Epigenetic Research, School of Dentistry, UCLA, 10833 Le Conte Avenue, 73-041 CHS, Los Angeles, CA 90095, USA; UCLA Broad Stem Cell Research Center, Box 957357, Los Angeles, CA 90095, USA; UCLA's Jonsson Comprehensive Cancer Center, 8-684 Factor Building, Box 951781, Los Angeles, CA 90095, USA. Electronic address: thadyk@ucla.edu.
Yong
Y
Kim
D000328:Adult; D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D019175:DNA Methylation / Q000187:drug effects; D003782:Dental Pulp / Q000166:cytology; D000431:Ethanol / Q000494:pharmacology*; D015870:Gene Expression / Q000187:drug effects*; D020869:Gene Expression Profiling; D006801:Humans; D056484:Jumonji Domain-Containing Histone Demethylases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008819:Mice, Nude; D009805:Odontogenesis / Q000187:drug effects; D010012:Osteogenesis / Q000187:drug effects; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*
D034741:RNA, Small Interfering; D000431:Ethanol; D056484:Jumonji Domain-Containing Histone Demethylases; C524220:KDM6B protein, human; D000469:Alkaline Phosphatase
Alcohol; DNA methylation; Dental pulp stem cells (DPSCs); Epigenetics; Mineralization; Odontogenic differentiation; Osteogenic differentiation