Whole-transcriptome analysis of endothelial to hematopoietic stem cell transition reveals a requirement for Gpr56 in HSC generation

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]),...

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Published inThe Journal of experimental medicine Vol. 212; no. 1; pp. 93 - 106
Main Authors Solaimani Kartalaei, Parham, Yamada-Inagawa, Tomoko, Vink, Chris S, de Pater, Emma, van der Linden, Reinier, Marks-Bluth, Jonathon, van der Sloot, Anthon, van den Hout, Mirjam, Yokomizo, Tomomasa, van Schaick-Solernó, M Lucila, Delwel, Ruud, Pimanda, John E, van IJcken, Wilfred F J, Dzierzak, Elaine
Format Journal Article
LanguageEnglish
Published United States The Rockefeller University Press 12.01.2015
Subjects
Animals
Cell Line
Cell Transdifferentiation - genetics
Cells, Cultured
Cercopithecus aethiops
CHO Cells
COS Cells
Cricetinae
Cricetulus
Embryo, Mammalian - cytology
Embryo, Mammalian - metabolism
Endothelial Cells - cytology
Endothelial Cells - metabolism
Female
Gene Expression Profiling - methods
Gene Ontology
Hematopoietic Stem Cells - cytology
Hematopoietic Stem Cells - metabolism
Humans
In Situ Hybridization
Mice, Inbred C57BL
Microscopy, Confocal
Oligonucleotide Array Sequence Analysis
Receptors, G-Protein-Coupled - genetics
Reverse Transcriptase Polymerase Chain Reaction
Sequence Analysis, RNA - methods
Up-Regulation
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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.
AbstractList Using highly sensitive RNAseq to examine the whole transcriptome of enriched aortic hematopoietic stem cells and endothelial cells, the authors find G-protein–coupled receptor, Gpr56, is required to generate the first HSCs during endothelial to hematopoietic cell transition. 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.
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.
Using highly sensitive RNAseq to examine the whole transcriptome of enriched aortic hematopoietic stem cells and endothelial cells, the authors find G-protein-coupled receptor, Gpr56, is required to generate the first HSCs during endothelial to hematopoietic cell transition. 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.
Author Solaimani Kartalaei, Parham
Dzierzak, Elaine
van der Linden, Reinier
de Pater, Emma
van IJcken, Wilfred F J
Marks-Bluth, Jonathon
Vink, Chris S
van der Sloot, Anthon
Yokomizo, Tomomasa
Delwel, Ruud
van den Hout, Mirjam
Pimanda, John E
van Schaick-Solernó, M Lucila
Yamada-Inagawa, Tomoko
AuthorAffiliation 4 Lowy Cancer Research Centre and Prince of Wales Clinical School, University of New South Wales, Sydney, New South Wales 2052, Australia
1 Erasmus MC Stem Cell Institute, Department of Cell Biology , 2 Center for Biomics , and 3 Department of Hematology, Erasmus University Medical Center, 3000 CA Rotterdam, Netherlands
5 Cancer Science Institute of Singapore, National University of Singapore, Singapore 117599
AuthorAffiliation_xml – name: 1 Erasmus MC Stem Cell Institute, Department of Cell Biology , 2 Center for Biomics , and 3 Department of Hematology, Erasmus University Medical Center, 3000 CA Rotterdam, Netherlands
– name: 5 Cancer Science Institute of Singapore, National University of Singapore, Singapore 117599
– name: 4 Lowy Cancer Research Centre and Prince of Wales Clinical School, University of New South Wales, Sydney, New South Wales 2052, Australia
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  givenname: Parham
  surname: Solaimani Kartalaei
  fullname: Solaimani Kartalaei, Parham
  organization: Erasmus MC Stem Cell Institute, Department of Cell Biology, Center for Biomics, and Department of Hematology, Erasmus University Medical Center, 3000 CA Rotterdam, Netherlands
– sequence: 2
  givenname: Tomoko
  surname: Yamada-Inagawa
  fullname: Yamada-Inagawa, Tomoko
  organization: Erasmus MC Stem Cell Institute, Department of Cell Biology, Center for Biomics, and Department of Hematology, Erasmus University Medical Center, 3000 CA Rotterdam, Netherlands
– sequence: 3
  givenname: Chris S
  surname: Vink
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  givenname: Emma
  surname: de Pater
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  organization: Erasmus MC Stem Cell Institute, Department of Cell Biology, Center for Biomics, and Department of Hematology, Erasmus University Medical Center, 3000 CA Rotterdam, Netherlands
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  organization: Erasmus MC Stem Cell Institute, Department of Cell Biology, Center for Biomics, and Department of Hematology, Erasmus University Medical Center, 3000 CA Rotterdam, Netherlands
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  surname: Marks-Bluth
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  organization: Lowy Cancer Research Centre and Prince of Wales Clinical School, University of New South Wales, Sydney, New South Wales 2052, Australia
– sequence: 7
  givenname: Anthon
  surname: van der Sloot
  fullname: van der Sloot, Anthon
  organization: Erasmus MC Stem Cell Institute, Department of Cell Biology, Center for Biomics, and Department of Hematology, Erasmus University Medical Center, 3000 CA Rotterdam, Netherlands
– sequence: 8
  givenname: Mirjam
  surname: van den Hout
  fullname: van den Hout, Mirjam
  organization: Erasmus MC Stem Cell Institute, Department of Cell Biology, Center for Biomics, and Department of Hematology, Erasmus University Medical Center, 3000 CA Rotterdam, Netherlands
– sequence: 9
  givenname: Tomomasa
  surname: Yokomizo
  fullname: Yokomizo, Tomomasa
  organization: Cancer Science Institute of Singapore, National University of Singapore, Singapore 117599
– sequence: 10
  givenname: M Lucila
  surname: van Schaick-Solernó
  fullname: van Schaick-Solernó, M Lucila
  organization: Erasmus MC Stem Cell Institute, Department of Cell Biology, Center for Biomics, and Department of Hematology, Erasmus University Medical Center, 3000 CA Rotterdam, Netherlands
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  givenname: Ruud
  surname: Delwel
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  organization: Lowy Cancer Research Centre and Prince of Wales Clinical School, University of New South Wales, Sydney, New South Wales 2052, Australia
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  givenname: Wilfred F J
  surname: van IJcken
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  organization: Erasmus MC Stem Cell Institute, Department of Cell Biology, Center for Biomics, and Department of Hematology, Erasmus University Medical Center, 3000 CA Rotterdam, Netherlands
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  givenname: Elaine
  surname: Dzierzak
  fullname: Dzierzak, Elaine
  email: e.dzierzak@erasmusmc.nl, Elaine.Dzierzak@ed.ac.uk
  organization: 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
BackLink https://www.ncbi.nlm.nih.gov/pubmed/25547674$$D View this record in MEDLINE/PubMed
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content type line 23
E. Dzierzak’s present address is University of Edinburgh/Medical Research Council Centre for Inflammation Research, Queens Medical Research Institute, Edinburgh EH16 4TJ, Scotland, UK.
P. Solaimani Kartalaei and T. Yamada-Inagawa contributed equally to this paper.
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SSID ssj0014456
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Snippet Hematopoietic stem cells (HSCs) are generated via a natural transdifferentiation process known as endothelial to hematopoietic cell transition (EHT). Because...
Using highly sensitive RNAseq to examine the whole transcriptome of enriched aortic hematopoietic stem cells and endothelial cells, the authors find...
SourceID pubmedcentral
proquest
crossref
pubmed
SourceType Open Access Repository
Aggregation Database
Index Database
StartPage 93
SubjectTerms Animals
Cell Line
Cell Transdifferentiation - genetics
Cells, Cultured
Cercopithecus aethiops
CHO Cells
COS Cells
Cricetinae
Cricetulus
Embryo, Mammalian - cytology
Embryo, Mammalian - metabolism
Endothelial Cells - cytology
Endothelial Cells - metabolism
Female
Gene Expression Profiling - methods
Gene Ontology
Hematopoietic Stem Cells - cytology
Hematopoietic Stem Cells - metabolism
Humans
In Situ Hybridization
Mice, Inbred C57BL
Microscopy, Confocal
Oligonucleotide Array Sequence Analysis
Receptors, G-Protein-Coupled - genetics
Reverse Transcriptase Polymerase Chain Reaction
Sequence Analysis, RNA - methods
Up-Regulation
Title Whole-transcriptome analysis of endothelial to hematopoietic stem cell transition reveals a requirement for Gpr56 in HSC generation
URI https://www.ncbi.nlm.nih.gov/pubmed/25547674
https://search.proquest.com/docview/1660028027
https://search.proquest.com/docview/1808699949
https://pubmed.ncbi.nlm.nih.gov/PMC4291529
Volume 212
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