C/EBPα creates elite cells for iPSC reprogramming by upregulating Klf4 and increasing the levels of Lsd1 and Brd4

Bruno Di Stefano; Samuel Collombet; Janus Schou Jakobsen; Michael Wierer; Jose Luis Sardina; Andreas Lackner; Ralph Stadhouders; Carolina Segura-Morales; Mirko Francesconi; Francesco Limone; Matthias Mann; Bo Porse; Denis Thieffry; Thomas Graf

doi:10.1038/ncb3326

C/EBPα creates elite cells for iPSC reprogramming by upregulating Klf4 and increasing the levels of Lsd1 and Brd4

Nat Cell Biol. 2016 Apr;18(4):371-81. doi: 10.1038/ncb3326. Epub 2016 Mar 14.

Authors

Bruno Di Stefano^{1

2}, Samuel Collombet³, Janus Schou Jakobsen^{4

5

6}, Michael Wierer⁷, Jose Luis Sardina^{1

2}, Andreas Lackner^{1

2}, Ralph Stadhouders^{1

2}, Carolina Segura-Morales^{1

2}, Mirko Francesconi^{1

2}, Francesco Limone^{1

2}, Matthias Mann⁷, Bo Porse^{4

5

6}, Denis Thieffry³, Thomas Graf^{1

2}

Affiliations

¹ Centre for Genomic Regulation (CRG), The Barcelona Institute of Science and Technology, Dr. Aiguader 88, Barcelona 08003, Spain.
² Universitat Pompeu Fabra (UPF), Barcelona 08003, Spain.
³ Institut de Biologie de l'Ecole Normale Supérieure (IBENS), CNRS UMR8197, INSERM U1024, Ecole Normale Supérieure, PSL Research University, F-75005 Paris, France.
⁴ The Finsen Laboratory, Rigshospitalet, Faculty of Health Sciences, University of Copenhagen, Ole Maaløes Vej 5, Copenhagen 2200, Denmark.
⁵ Biotech Research and Innovation Centre (BRIC), University of Copenhagen, Ole Maaløes Vej 5, Copenhagen 2200, Denmark.
⁶ Danish Stem Cell Center (DanStem), Faculty of Health Sciences, University of Copenhagen, 3B Blegdamsvej, Copenhagen 2200, Denmark.
⁷ Max Planck Institute of Biochemistry, 82152 Munich, Germany.

PMID: 26974661
DOI: 10.1038/ncb3326

Abstract

Reprogramming somatic cells into induced pluripotent stem cells (iPSCs) is typically inefficient and has been explained by elite-cell and stochastic models. We recently reported that B cells exposed to a pulse of C/EBPα (Bα' cells) behave as elite cells, in that they can be rapidly and efficiently reprogrammed into iPSCs by the Yamanaka factors OSKM. Here we show that C/EBPα post-transcriptionally increases the abundance of several hundred proteins, including Lsd1, Hdac1, Brd4, Med1 and Cdk9, components of chromatin-modifying complexes present at super-enhancers. Lsd1 was found to be required for B cell gene silencing and Brd4 for the activation of the pluripotency program. C/EBPα also promotes chromatin accessibility in pluripotent cells and upregulates Klf4 by binding to two haematopoietic enhancers. Bα' cells share many properties with granulocyte/macrophage progenitors, naturally occurring elite cells that are obligate targets for leukaemic transformation, whose formation strictly requires C/EBPα.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
B-Lymphocytes / metabolism
Blotting, Western
CCAAT-Enhancer-Binding Protein-alpha / genetics*
CCAAT-Enhancer-Binding Protein-alpha / metabolism
Cell Line
Cells, Cultured
Cellular Reprogramming / genetics*
Female
Gene Expression Profiling / methods
Gene Ontology
HEK293 Cells
Histone Demethylases / genetics*
Histone Demethylases / metabolism
Humans
Induced Pluripotent Stem Cells / metabolism*
Kruppel-Like Factor 4
Kruppel-Like Transcription Factors / genetics*
Kruppel-Like Transcription Factors / metabolism
Male
Mice
Mice, Inbred C57BL
Mouse Embryonic Stem Cells / metabolism
Nuclear Proteins / genetics*
Nuclear Proteins / metabolism
Proteomics / methods
Reverse Transcriptase Polymerase Chain Reaction
Transcription Factors / genetics*
Transcription Factors / metabolism
Up-Regulation

Substances

Brd4 protein, mouse
CCAAT-Enhancer-Binding Protein-alpha
KLF4 protein, human
Klf4 protein, mouse
Kruppel-Like Factor 4
Kruppel-Like Transcription Factors
Nuclear Proteins
Transcription Factors
Histone Demethylases
KDM1a protein, mouse