title: An autonomous CEBPA enhancer specific for myeloid-lineage priming and neutrophilic differentiation.
abstract: Neutrophilic differentiation is dependent on CCAAT enhancer-binding protein α (C/EBPα), a transcription factor expressed in multiple organs including the bone marrow. Using functional genomic technologies in combination with clustered regularly-interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 genome editing and in vivo mouse modeling, we show that CEBPA is located in a 170-kb topological-associated domain that contains 14 potential enhancers. Of these, 1 enhancer located +42 kb from CEBPA is active and engages with the CEBPA promoter in myeloid cells only. Germ line deletion of the homologous enhancer in mice in vivo reduces Cebpa levels exclusively in hematopoietic stem cells (HSCs) and myeloid-primed progenitor cells leading to severe defects in the granulocytic lineage, without affecting any other Cebpa-expressing organ studied. The enhancer-deleted progenitor cells lose their myeloid transcription program and are blocked in differentiation. Deletion of the enhancer also causes loss of HSC maintenance. We conclude that a single +42-kb enhancer is essential for CEBPA expression in myeloid cells only.
authors.affiliation: Department of Hematology.
authors.forename: Roberto
authors.initials: R
authors.lastname: Avellino
authors.affiliation: Department of Hematology.
authors.forename: Marije
authors.initials: M
authors.lastname: Havermans
authors.affiliation: Department of Hematology.
authors.forename: Claudia
authors.initials: C
authors.lastname: Erpelinck
authors.affiliation: Department of Hematology.
authors.forename: Mathijs A
authors.initials: MA
authors.lastname: Sanders
authors.affiliation: Department of Hematology.
authors.forename: Remco
authors.initials: R
authors.lastname: Hoogenboezem
authors.affiliation: Cancer Computational Biology Center, Department of Cell Biology, and Department of Urology, Erasmus University Medical Center, Rotterdam, The Netherlands;
authors.forename: Harmen J G
authors.identifier: 0000-0002-9794-1477
authors.initials: HJ
authors.lastname: van de Werken
authors.affiliation: Department of Hematology.
authors.forename: Elwin
authors.initials: E
authors.lastname: Rombouts
authors.affiliation: Department of Hematology.
authors.forename: Kirsten
authors.initials: K
authors.lastname: van Lom
authors.affiliation: Department of Hematology.
authors.forename: Paulina M H
authors.initials: PM
authors.lastname: van Strien
authors.affiliation: Department of Internal Medicine III and Regensburg Centre for Interventional Immunology, University Hospital Regensburg, Regensburg, Germany;
authors.forename: Claudia
authors.initials: C
authors.lastname: Gebhard
authors.affiliation: Department of Internal Medicine III and Regensburg Centre for Interventional Immunology, University Hospital Regensburg, Regensburg, Germany;
authors.forename: Michael
authors.initials: M
authors.lastname: Rehli
authors.affiliation: Prince of Wales Clinical Schools and Lowy Cancer Research Centre, The University of New South Wales, Sydney, NSW, Australia;
authors.forename: John
authors.initials: J
authors.lastname: Pimanda
authors.affiliation: Prince of Wales Clinical Schools and Lowy Cancer Research Centre, The University of New South Wales, Sydney, NSW, Australia;
authors.forename: Dominik
authors.initials: D
authors.lastname: Beck
authors.affiliation: Department of Hematology, Department of Immunology and.
authors.forename: Stefan
authors.initials: S
authors.lastname: Erkeland
authors.affiliation: Department of Viroscience, Erasmus University Medical Center, Rotterdam, The Netherlands; and.
authors.forename: Thijs
authors.initials: T
authors.lastname: Kuiken
authors.affiliation: Department of Hematology.
authors.forename: Hans
authors.initials: H
authors.lastname: de Looper
authors.affiliation: Department of Hematology, Department of Translational Oncology, National Center for Tumor Diseases, Deutsches Krebsforschungszentrum (DKFZ), Heidelberg, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Gröschel
authors.affiliation: Department of Hematology.
authors.forename: Ivo
authors.initials: I
authors.lastname: Touw
authors.affiliation: Department of Hematology.
authors.forename: Eric
authors.initials: E
authors.lastname: Bindels
authors.affiliation: Department of Hematology.
authors.forename: Ruud
authors.initials: R
authors.lastname: Delwel
mesh_terms: D000818:Animals; D022763:CCAAT-Enhancer-Binding Protein-alpha / Q000235:genetics / Q000378:metabolism*; D002454:Cell Differentiation / Q000235:genetics*; D045744:Cell Line, Tumor; D019070:Cell Lineage / Q000235:genetics*; D004742:Enhancer Elements, Genetic; D018507:Gene Expression Regulation, Developmental; D057809:HEK293 Cells; D018922:HL-60 Cells; D006367:HeLa Cells; D056945:Hep G2 Cells; D006801:Humans; D019169:Jurkat Cells; D020014:K562 Cells; D051379:Mice; D018345:Mice, Knockout; D022423:Myeloid Cells / Q000502:physiology*; D038042:Myelopoiesis / Q000235:genetics*; D009504:Neutrophils / Q000502:physiology*; D020298:U937 Cells
chemical_list: D022763:CCAAT-Enhancer-Binding Protein-alpha