title: Early dynamic fate changes in haemogenic endothelium characterized at the single-cell level.
abstract: Haematopoietic stem cells (HSCs) are the founding cells of the adult haematopoietic system, born during ontogeny from a specialized subset of endothelium, the haemogenic endothelium (HE) via an endothelial-to-haematopoietic transition (EHT). Although recently imaged in real time, the underlying mechanism of EHT is still poorly understood. We have generated a Runx1 +23 enhancer-reporter transgenic mouse (23GFP) for the prospective isolation of HE throughout embryonic development. Here we perform functional analysis of over 1,800 and transcriptional analysis of 268 single 23GFP(+) HE cells to explore the onset of EHT at the single-cell level. We show that initiation of the haematopoietic programme occurs in cells still embedded in the endothelial layer, and is accompanied by a previously unrecognized early loss of endothelial potential before HSCs emerge. Our data therefore provide important insights on the timeline of early haematopoietic commitment.
authors: MRC Molecular Haematology Unit, Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine, John Radcliffe Hospital, University of Oxford, Oxford OX3 9DS, UK.
authors: Gemma
authors: G
authors: Swiers
authors: MRC Molecular Haematology Unit, Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine, John Radcliffe Hospital, University of Oxford, Oxford OX3 9DS, UK.
authors: Claudia
authors: C
authors: Baumann
authors: MRC Molecular Haematology Unit, Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine, John Radcliffe Hospital, University of Oxford, Oxford OX3 9DS, UK.
authors: John
authors: J
authors: O'Rourke
authors: Computational Biology Research Group, Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine, John Radcliffe Hospital, University of Oxford, Oxford OX3 9DS, UK.
authors: Eleni
authors: E
authors: Giannoulatou
authors: Computational Biology Research Group, Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine, John Radcliffe Hospital, University of Oxford, Oxford OX3 9DS, UK.
authors: Stephen
authors: S
authors: Taylor
authors: Department of Haematology, Cambridge Institute for Medical Research and Wellcome Trust & MRC Cambridge Stem Cell Institute, Cambridge CB2 OXY, UK.
authors: Anagha
authors: A
authors: Joshi
authors: Department of Haematology, Cambridge Institute for Medical Research and Wellcome Trust & MRC Cambridge Stem Cell Institute, Cambridge CB2 OXY, UK.
authors: Victoria
authors: V
authors: Moignard
authors: MRC Molecular Haematology Unit, Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine, John Radcliffe Hospital, University of Oxford, Oxford OX3 9DS, UK.
authors: Cristina
authors: C
authors: Pina
authors: MRC Molecular Haematology Unit, Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine, John Radcliffe Hospital, University of Oxford, Oxford OX3 9DS, UK.
authors: Thomas
authors: T
authors: Bee
authors: Research Unit Stem Cell Dynamics, Helmholtz Center Munich-German Research Center for Environmental Health, Ingolstaedter Landstrasse 1 85764 Neuherberg, Germany.
authors: Konstantinos D
authors: KD
authors: Kokkaliaris
authors: Wells Center for Pediatric Research, Department of Pediatrics, Indiana University School of Medicine, Indianapolis, Indiana 46202, USA.
authors: Momoko
authors: M
authors: Yoshimoto
authors: Wells Center for Pediatric Research, Department of Pediatrics, Indiana University School of Medicine, Indianapolis, Indiana 46202, USA.
authors: Mervin C
authors: MC
authors: Yoder
authors: Institute for Biomedical Research, College of Medical and Dental Sciences, University of Birmingham, Birmingham B15 2TT, UK.
authors: Jon
authors: J
authors: Frampton
authors: Research Unit Stem Cell Dynamics, Helmholtz Center Munich-German Research Center for Environmental Health, Ingolstaedter Landstrasse 1 85764 Neuherberg, Germany.
authors: Timm
authors: T
authors: Schroeder
authors: MRC Molecular Haematology Unit, Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine, John Radcliffe Hospital, University of Oxford, Oxford OX3 9DS, UK.
authors: Tariq
authors: T
authors: Enver
authors: Department of Haematology, Cambridge Institute for Medical Research and Wellcome Trust & MRC Cambridge Stem Cell Institute, Cambridge CB2 OXY, UK.
authors: Berthold
authors: B
authors: Göttgens
authors: MRC Molecular Haematology Unit, Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine, John Radcliffe Hospital, University of Oxford, Oxford OX3 9DS, UK.
authors: Marella F T R
authors: MFTR
authors: de Bruijn
mesh_terms: D000818:Animals; D050676:Core Binding Factor Alpha 2 Subunit / Q000235:genetics; D004622:Embryo, Mammalian / Q000166:cytology*; D004742:Enhancer Elements, Genetic; D005260:Female; D018507:Gene Expression Regulation, Developmental; D049452:Green Fluorescent Proteins / Q000235:genetics; D055018:Hemangioblasts / Q000166:cytology* / Q000502:physiology*; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D011247:Pregnancy; D059010:Single-Cell Analysis / Q000379:methods*
chemical_list: D050676:Core Binding Factor Alpha 2 Subunit; C493726:Runx1 protein, mouse; D049452:Green Fluorescent Proteins