title: Induced DNA demethylation by targeting Ten-Eleven Translocation 2 to the human ICAM-1 promoter.
abstract: Increasing evidence indicates that active DNA demethylation is involved in several processes in mammals, resulting in developmental stage-specificity and cell lineage-specificity. The recently discovered Ten-Eleven Translocation (TET) dioxygenases are accepted to be involved in DNA demethylation by initiating 5-mC oxidation. Aberrant DNA methylation profiles are associated with many diseases. For example in cancer, hypermethylation results in silencing of tumor suppressor genes. Such silenced genes can be re-expressed by epigenetic drugs, but this approach has genome-wide effects. In this study, fusions of designer DNA binding domains to TET dioxygenase family members (TET1, -2 or -3) were engineered to target epigenetically silenced genes (ICAM-1, EpCAM). The effects on targeted CpGs' methylation and on expression levels of the target genes were assessed. The results indicated demethylation of targeted CpG sites in both promoters for targeted TET2 and to a lesser extent for TET1, but not for TET3. Interestingly, we observed re-activation of transcription of ICAM-1. Thus, our work suggests that we provided a mechanism to induce targeted DNA demethylation, which facilitates re-activation of expression of the target genes. Furthermore, this Epigenetic Editing approach is a powerful tool to investigate functions of epigenetic writers and erasers and to elucidate consequences of epigenetic marks.
authors.affiliation: Epigenetic Editing, Department of Pathology and Medical Biology, University Medical Center Groningen, University of Groningen, Hanzeplein1, 9713 GZ Groningen, The Netherlands, The State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, 320 Yueyang Road, Shanghai 200031, China and Synvolux Therapeutics Inc., LJ. Zielstraweg 1, 9713 GX Groningen, The Netherlands.
authors.forename: Hui
authors.initials: H
authors.lastname: Chen
authors.forename: Hinke G
authors.initials: HG
authors.lastname: Kazemier
authors.forename: Marloes L
authors.initials: ML
authors.lastname: de Groote
authors.forename: Marcel H J
authors.initials: MH
authors.lastname: Ruiters
authors.forename: Guo-Liang
authors.initials: GL
authors.lastname: Xu
authors.forename: Marianne G
authors.initials: MG
authors.lastname: Rots
mesh_terms: D000818:Animals; D000951:Antigens, Neoplasm / Q000235:genetics; D015815:Cell Adhesion Molecules / Q000235:genetics; D045744:Cell Line, Tumor; D004247:DNA / Q000378:metabolism*; D019175:DNA Methylation; D004268:DNA-Binding Proteins / Q000378:metabolism*; D049308:Dioxygenases / Q000378:metabolism*; D044127:Epigenesis, Genetic; D000071858:Epithelial Cell Adhesion Molecule; D057809:HEK293 Cells; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000096:biosynthesis / Q000235:genetics*; D051379:Mice; D011401:Promoter Regions, Genetic; D011518:Proto-Oncogene Proteins / Q000378:metabolism*; D015533:Transcriptional Activation; D016335:Zinc Fingers
chemical_list: D000951:Antigens, Neoplasm; D015815:Cell Adhesion Molecules; D004268:DNA-Binding Proteins; D000071858:Epithelial Cell Adhesion Molecule; D011518:Proto-Oncogene Proteins; D018799:Intercellular Adhesion Molecule-1; D004247:DNA; D049308:Dioxygenases; C551914:Tet2 protein, mouse