title: C/EBPα poises B cells for rapid reprogramming into induced pluripotent stem cells.
abstract: CCAAT/enhancer binding protein-α (C/EBPα) induces transdifferentiation of B cells into macrophages at high efficiencies and enhances reprogramming into induced pluripotent stem (iPS) cells when co-expressed with the transcription factors Oct4 (Pou5f1), Sox2, Klf4 and Myc (hereafter called OSKM). However, how C/EBPα accomplishes these effects is unclear. Here we find that in mouse primary B cells transient C/EBPα expression followed by OSKM activation induces a 100-fold increase in iPS cell reprogramming efficiency, involving 95% of the population. During this conversion, pluripotency and epithelial-mesenchymal transition genes become markedly upregulated, and 60% of the cells express Oct4 within 2 days. C/EBPα acts as a 'path-breaker' as it transiently makes the chromatin of pluripotency genes more accessible to DNase I. C/EBPα also induces the expression of the dioxygenase Tet2 and promotes its translocation to the nucleus where it binds to regulatory regions of pluripotency genes that become demethylated after OSKM induction. In line with these findings, overexpression of Tet2 enhances OSKM-induced B-cell reprogramming. Because the enzyme is also required for efficient C/EBPα-induced immune cell conversion, our data indicate that Tet2 provides a mechanistic link between iPS cell reprogramming and B-cell transdifferentiation. The rapid iPS reprogramming approach described here should help to fully elucidate the process and has potential clinical applications.
authors: 1] Gene Regulation, Stem Cells and Cancer Programme, Centre for Genomic Regulation (CRG), Dr Aiguader 88, 08003 Barcelona, Spain [2] Universitat Pompeu Fabra (UPF), Dr Aiguader 88, 08003 Barcelona, Spain.
authors: Bruno
authors: B
authors: Di Stefano
authors: 1] Gene Regulation, Stem Cells and Cancer Programme, Centre for Genomic Regulation (CRG), Dr Aiguader 88, 08003 Barcelona, Spain [2] Universitat Pompeu Fabra (UPF), Dr Aiguader 88, 08003 Barcelona, Spain [3].
authors: Jose Luis
authors: JL
authors: Sardina
authors: 1] Gene Regulation, Stem Cells and Cancer Programme, Centre for Genomic Regulation (CRG), Dr Aiguader 88, 08003 Barcelona, Spain [2] Universitat Pompeu Fabra (UPF), Dr Aiguader 88, 08003 Barcelona, Spain [3].
authors: Chris
authors: C
authors: van Oevelen
authors: 1] Ecole Normale Supérieure, Institut de Biologie de l'ENS, 45 Rue d'Ulm, Paris F-75005, France [2] Inserm, U1024, Paris F-75005, France [3] CNRS, UMR 8197, Paris F-75005, France.
authors: Samuel
authors: S
authors: Collombet
authors: 1] Gene Regulation, Stem Cells and Cancer Programme, Centre for Genomic Regulation (CRG), Dr Aiguader 88, 08003 Barcelona, Spain [2] Universitat Pompeu Fabra (UPF), Dr Aiguader 88, 08003 Barcelona, Spain [3] Memorial Sloan Kettering Cancer Center, 1275 York Avenue, New York 10065, USA.
authors: Eric M
authors: EM
authors: Kallin
authors: 1] Gene Regulation, Stem Cells and Cancer Programme, Centre for Genomic Regulation (CRG), Dr Aiguader 88, 08003 Barcelona, Spain [2] Universitat Pompeu Fabra (UPF), Dr Aiguader 88, 08003 Barcelona, Spain.
authors: Guillermo P
authors: GP
authors: Vicent
authors: Yale Cancer Center, Yale University School of Medicine, 333 Cedar Street, New Haven, Connecticut 06510, USA.
authors: Jun
authors: J
authors: Lu
authors: 1] Ecole Normale Supérieure, Institut de Biologie de l'ENS, 45 Rue d'Ulm, Paris F-75005, France [2] Inserm, U1024, Paris F-75005, France [3] CNRS, UMR 8197, Paris F-75005, France.
authors: Denis
authors: D
authors: Thieffry
authors: 1] Gene Regulation, Stem Cells and Cancer Programme, Centre for Genomic Regulation (CRG), Dr Aiguader 88, 08003 Barcelona, Spain [2] Universitat Pompeu Fabra (UPF), Dr Aiguader 88, 08003 Barcelona, Spain.
authors: Miguel
authors: M
authors: Beato
authors: 1] Gene Regulation, Stem Cells and Cancer Programme, Centre for Genomic Regulation (CRG), Dr Aiguader 88, 08003 Barcelona, Spain [2] Universitat Pompeu Fabra (UPF), Dr Aiguader 88, 08003 Barcelona, Spain [3] Institució Catalana de Recerca i Estudis Avançats (ICREA), Pg Lluis Companys 23, 08010 Barcelona, Spain.
authors: Thomas
authors: T
authors: Graf
mesh_terms: D000818:Animals; D001402:B-Lymphocytes / Q000166:cytology* / Q000378:metabolism*; D022763:CCAAT-Enhancer-Binding Protein-alpha / Q000235:genetics / Q000378:metabolism*; D054338:Cell Transdifferentiation; D002478:Cells, Cultured; D065150:Cellular Reprogramming / Q000235:genetics; D002843:Chromatin / Q000235:genetics / Q000378:metabolism; D003596:Cytosine / Q000378:metabolism; D019175:DNA Methylation; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D003850:Deoxyribonuclease I / Q000378:metabolism; D049308:Dioxygenases; D058750:Epithelial-Mesenchymal Transition / Q000235:genetics; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism*; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors / Q000235:genetics / Q000378:metabolism; D051379:Mice; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics / Q000378:metabolism; D055748:SOXB1 Transcription Factors / Q000235:genetics / Q000378:metabolism; D015854:Up-Regulation / Q000235:genetics
chemical_list: D022763:CCAAT-Enhancer-Binding Protein-alpha; D002843:Chromatin; D004268:DNA-Binding Proteins; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors; D050814:Octamer Transcription Factor-3; D011518:Proto-Oncogene Proteins; D016271:Proto-Oncogene Proteins c-myc; D055748:SOXB1 Transcription Factors; D003596:Cytosine; D049308:Dioxygenases; C551914:Tet2 protein, mouse; D003850:Deoxyribonuclease I