title: The regulatory role of histone deacetylase inhibitors in Fgf4 expression is dependent on the differentiation state of pluripotent stem cells.
abstract: The identity of embryonic stem cells (ESCs) is controlled by a set of pluripotency genes, including Oct4, Sox2, Nanog, and Fgf4. How their expression is repressed during differentiation and reactivated during reprogramming is largely unknown. Here, using mouse ESCs as well as F9 and P19 cells (mouse embryonal carcinoma cell lines, P19 being considered further differentiated than F9 cells) as models, we found that HDAC inhibitors elevated Fgf4 expression in P19 cells, but reduced it in F9 cells. We also observed that HDAC inhibitors enhanced the expression of Fgf4 and a subset of pluripotency genes in differentiated ESCs, but reduced their expression in undifferentiated and less differentiated ESCs. Mechanistically, we observed more HDAC1 recruitment and a weaker association of histone 4 lysine 5 acetylation at the Fgf4 enhancer in P19 cells compared to F9 cells. Additionally, we demonstrated the interaction between Sox2 and HDAC1 both in vitro and in vivo, implicating a possible role for Sox2 in the recruitment of HDAC1 to the Fgf4 enhancer. We also found that Nanog bound to the Fgf4 enhancer, and this binding was stronger in F9 cells, indicating the involvement of Nanog in the regulation of Fgf4 expression in undifferentiated and less differentiated pluripotent stem cells. This study uncovers an important role of HDAC1 and histone modifications in the repression of Fgf4 and perhaps other pluripotency genes during ESC differentiation. Our results also suggest that HDAC inhibitors may promote reprogramming partially through activating pluripotency genes at some intermediate stages.
authors.affiliation: Key Laboratory of Stem Cell Biology, Institute of Health Sciences, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences/Shanghai JiaoTong University School of Medicine, Shanghai, China.
authors.forename: Guilai
authors.initials: G
authors.lastname: Shi
authors.forename: Furong
authors.initials: F
authors.lastname: Gao
authors.forename: Ying
authors.initials: Y
authors.lastname: Jin
mesh_terms: D000107:Acetylation; D000818:Animals; D001665:Binding Sites; D002454:Cell Differentiation; D002460:Cell Line; D042002:Chromatin Assembly and Disassembly / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D054278:Embryonal Carcinoma Stem Cells / Q000187:drug effects* / Q000201:enzymology; D053595:Embryonic Stem Cells / Q000187:drug effects* / Q000201:enzymology; D004742:Enhancer Elements, Genetic; D051519:Fibroblast Growth Factor 4 / Q000235:genetics / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D056284:Histone Deacetylase 1 / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology*; D006657:Histones / Q000378:metabolism; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D006877:Hydroxamic Acids / Q000494:pharmacology; D051379:Mice; D000071317:Nanog Homeobox Protein; D039904:Pluripotent Stem Cells / Q000187:drug effects* / Q000201:enzymology; D034622:RNA Interference; D055748:SOXB1 Transcription Factors / Q000378:metabolism; D013997:Time Factors; D014162:Transfection; D014635:Valproic Acid / Q000494:pharmacology
chemical_list: C496446:Fgf4 protein, mouse; D051519:Fibroblast Growth Factor 4; D056572:Histone Deacetylase Inhibitors; D006657:Histones; D018398:Homeodomain Proteins; D006877:Hydroxamic Acids; D000071317:Nanog Homeobox Protein; C475540:Nanog protein, mouse; D055748:SOXB1 Transcription Factors; C527783:Sox2 protein, mouse; C012589:trichostatin A; D014635:Valproic Acid; C484389:Hdac1 protein, mouse; D056284:Histone Deacetylase 1