title: Whole-transcriptome analysis of endothelial to hematopoietic stem cell transition reveals a requirement for Gpr56 in HSC generation.
abstract: Hematopoietic stem cells (HSCs) are generated via a natural transdifferentiation process known as endothelial to hematopoietic cell transition (EHT). Because of small numbers of embryonal arterial cells undergoing EHT and the paucity of markers to enrich for hemogenic endothelial cells (ECs [HECs]), the genetic program driving HSC emergence is largely unknown. Here, we use a highly sensitive RNAseq method to examine the whole transcriptome of small numbers of enriched aortic HSCs, HECs, and ECs. Gpr56, a G-coupled protein receptor, is one of the most highly up-regulated of the 530 differentially expressed genes. Also, highly up-regulated are hematopoietic transcription factors, including the "heptad" complex of factors. We show that Gpr56 (mouse and human) is a target of the heptad complex and is required for hematopoietic cluster formation during EHT. Our results identify the processes and regulators involved in EHT and reveal the surprising requirement for Gpr56 in generating the first HSCs.
authors.affiliation: Erasmus MC Stem Cell Institute, Department of Cell Biology, Center for Biomics, and Department of Hematology, Erasmus University Medical Center, 3000 CA Rotterdam, Netherlands.
authors.forename: Parham
authors.initials: P
authors.lastname: Solaimani Kartalaei
authors.affiliation: Erasmus MC Stem Cell Institute, Department of Cell Biology, Center for Biomics, and Department of Hematology, Erasmus University Medical Center, 3000 CA Rotterdam, Netherlands.
authors.forename: Tomoko
authors.initials: T
authors.lastname: Yamada-Inagawa
authors.affiliation: Erasmus MC Stem Cell Institute, Department of Cell Biology, Center for Biomics, and Department of Hematology, Erasmus University Medical Center, 3000 CA Rotterdam, Netherlands.
authors.forename: Chris S
authors.initials: CS
authors.lastname: Vink
authors.affiliation: Erasmus MC Stem Cell Institute, Department of Cell Biology, Center for Biomics, and Department of Hematology, Erasmus University Medical Center, 3000 CA Rotterdam, Netherlands.
authors.forename: Emma
authors.initials: E
authors.lastname: de Pater
authors.affiliation: Erasmus MC Stem Cell Institute, Department of Cell Biology, Center for Biomics, and Department of Hematology, Erasmus University Medical Center, 3000 CA Rotterdam, Netherlands.
authors.forename: Reinier
authors.initials: R
authors.lastname: van der Linden
authors.affiliation: Lowy Cancer Research Centre and Prince of Wales Clinical School, University of New South Wales, Sydney, New South Wales 2052, Australia.
authors.forename: Jonathon
authors.initials: J
authors.lastname: Marks-Bluth
authors.affiliation: Erasmus MC Stem Cell Institute, Department of Cell Biology, Center for Biomics, and Department of Hematology, Erasmus University Medical Center, 3000 CA Rotterdam, Netherlands.
authors.forename: Anthon
authors.initials: A
authors.lastname: van der Sloot
authors.affiliation: Erasmus MC Stem Cell Institute, Department of Cell Biology, Center for Biomics, and Department of Hematology, Erasmus University Medical Center, 3000 CA Rotterdam, Netherlands.
authors.forename: Mirjam
authors.initials: M
authors.lastname: van den Hout
authors.affiliation: Cancer Science Institute of Singapore, National University of Singapore, Singapore 117599.
authors.forename: Tomomasa
authors.initials: T
authors.lastname: Yokomizo
authors.affiliation: Erasmus MC Stem Cell Institute, Department of Cell Biology, Center for Biomics, and Department of Hematology, Erasmus University Medical Center, 3000 CA Rotterdam, Netherlands.
authors.forename: M Lucila
authors.initials: ML
authors.lastname: van Schaick-Solernó
authors.affiliation: Erasmus MC Stem Cell Institute, Department of Cell Biology, Center for Biomics, and Department of Hematology, Erasmus University Medical Center, 3000 CA Rotterdam, Netherlands.
authors.forename: Ruud
authors.initials: R
authors.lastname: Delwel
authors.affiliation: Lowy Cancer Research Centre and Prince of Wales Clinical School, University of New South Wales, Sydney, New South Wales 2052, Australia.
authors.forename: John E
authors.initials: JE
authors.lastname: Pimanda
authors.affiliation: Erasmus MC Stem Cell Institute, Department of Cell Biology, Center for Biomics, and Department of Hematology, Erasmus University Medical Center, 3000 CA Rotterdam, Netherlands.
authors.forename: Wilfred F J
authors.initials: WF
authors.lastname: van IJcken
authors.affiliation: Erasmus MC Stem Cell Institute, Department of Cell Biology, Center for Biomics, and Department of Hematology, Erasmus University Medical Center, 3000 CA Rotterdam, Netherlands e.dzierzak@erasmusmc.nl Elaine.Dzierzak@ed.ac.uk.
authors.forename: Elaine
authors.initials: E
authors.lastname: Dzierzak
mesh_terms: D000818:Animals; D016466:CHO Cells; D019556:COS Cells; D002460:Cell Line; D054338:Cell Transdifferentiation / Q000235:genetics*; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D006224:Cricetinae; D003412:Cricetulus; D004622:Embryo, Mammalian / Q000166:cytology / Q000378:metabolism; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism*; D005260:Female; D020869:Gene Expression Profiling / Q000379:methods*; D063990:Gene Ontology; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D017403:In Situ Hybridization; D008810:Mice, Inbred C57BL; D018613:Microscopy, Confocal; D020411:Oligonucleotide Array Sequence Analysis; D043562:Receptors, G-Protein-Coupled / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017423:Sequence Analysis, RNA / Q000379:methods; D015854:Up-Regulation
chemical_list: D043562:Receptors, G-Protein-Coupled; C118925:ADGRG1 protein, human; C524781:Adgrg1 protein, mouse