title: 5-azacytidine affects TET2 and histone transcription and reshapes morphology of human skin fibroblasts.
abstract: Phenotype definition is controlled by epigenetic regulations that allow cells to acquire their differentiated state. The process is reversible and attractive for therapeutic intervention and for the reactivation of hypermethylated pluripotency genes that facilitate transition to a higher plasticity state. We report the results obtained in human fibroblasts exposed to the epigenetic modifier 5-azacytidine (5-aza-CR), which increases adult cell plasticity and facilitates phenotype change. Although many aspects controlling its demethylating action have been widely investigated, the mechanisms underlying 5-aza-CR effects on cell plasticity are still poorly understood. Our experiments confirm decreased global methylation, but also demonstrate an increase of both Formylcytosine (5fC) and 5-Carboxylcytosine (5caC), indicating 5-aza-CR ability to activate a direct and active demethylating effect, possibly mediated via TET2 protein increased transcription. This was accompanied by transient upregulation of pluripotency markers and incremented histone expression, paralleled by changes in histone acetylating enzymes. Furthermore, adult fibroblasts reshaped into undifferentiated progenitor-like phenotype, with a sparse and open chromatin structure. Our findings indicate that 5-aza-CR induced somatic cell transition to a higher plasticity state is activated by multiple regulations that accompany the demethylating effect exerted by the modifier.
authors.affiliation: Laboratory of Biomedical Embryology, Centre for Stem Cell Research; Università degli Studi di Milano, Milan, 20133, Italy.
authors.forename: Elena F M
authors.initials: EF
authors.lastname: Manzoni
authors.affiliation: Laboratory of Biomedical Embryology, Centre for Stem Cell Research; Università degli Studi di Milano, Milan, 20133, Italy.
authors.forename: Georgia
authors.initials: G
authors.lastname: Pennarossa
authors.affiliation: Department of Biotechnology and Life Sciences; Università degli Studi dell'Insubria, Varese, 21100, Italy.
authors.forename: Magda
authors.initials: M
authors.lastname: deEguileor
authors.affiliation: Department of Biotechnology and Life Sciences; Università degli Studi dell'Insubria, Varese, 21100, Italy.
authors.forename: Gianluca
authors.initials: G
authors.lastname: Tettamanti
authors.affiliation: Laboratory of Biomedical Embryology, Centre for Stem Cell Research; Università degli Studi di Milano, Milan, 20133, Italy.
authors.forename: Fulvio
authors.initials: F
authors.lastname: Gandolfi
authors.affiliation: Laboratory of Biomedical Embryology, Centre for Stem Cell Research; Università degli Studi di Milano, Milan, 20133, Italy.
authors.forename: Tiziana A L
authors.initials: TA
authors.lastname: Brevini
mesh_terms: D000328:Adult; D001374:Azacitidine / Q000494:pharmacology*; D002478:Cells, Cultured; D002843:Chromatin / Q000737:chemistry / Q000378:metabolism; D019175:DNA Methylation / Q000187:drug effects; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D049308:Dioxygenases; D015536:Down-Regulation / Q000187:drug effects; D044127:Epigenesis, Genetic / Q000187:drug effects; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D051548:Histone Acetyltransferases / Q000235:genetics / Q000378:metabolism; D056284:Histone Deacetylase 1 / Q000235:genetics / Q000378:metabolism; D006657:Histones / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008854:Microscopy, Electron; D008875:Middle Aged; D011518:Proto-Oncogene Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects*; D015854:Up-Regulation / Q000187:drug effects*
chemical_list: D002843:Chromatin; D004268:DNA-Binding Proteins; D006657:Histones; D011518:Proto-Oncogene Proteins; D034741:RNA, Small Interfering; D014157:Transcription Factors; D049308:Dioxygenases; C541081:TET2 protein, human; D051548:Histone Acetyltransferases; C110880:histone acetyltransferase type B complex; C110550:HDAC1 protein, human; D056284:Histone Deacetylase 1; D001374:Azacitidine