Single Cell Resolution of Human Hematoendothelial Cells Defines Transcriptional Signatures of Hemogenic Endothelium

Endothelial‐to‐hematopoietic transition (EHT) is an important stage in definitive hematopoietic development. However, the genetic mechanisms underlying human EHT remain poorly characterized. We performed single cell RNA‐seq using 55 hemogenic endothelial cells (HECs: CD31+CD144+CD41–CD43–CD45–CD73–R...

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Published inStem cells (Dayton, Ohio) Vol. 36; no. 2; pp. 206 - 217
Main Authors Angelos, Mathew G., Abrahante, Juan E., Blum, Robert H., Kaufman, Dan S.
Format Journal Article
LanguageEnglish
Published United States Oxford University Press 01.02.2018
Subjects
Antigens, Surface - metabolism
Biomarkers
CD34 antigen
CD43 antigen
CD45 antigen
CD73 antigen
Cell Differentiation - genetics
Cell Differentiation - physiology
Cells (biology)
Cells, Cultured
Computational Biology
Developmental stages
Embryo cells
Embryonic Stem Cells - cytology
Embryonic Stem Cells - metabolism
Embryos
Endothelial cells
Endothelium
Flow Cytometry
Hemangioblasts - cytology
Hemangioblasts - metabolism
Hematopoiesis - genetics
Hematopoiesis - physiology
Hematopoietic stem cells
Hematopoietic Stem Cells - cytology
Hematopoietic Stem Cells - metabolism
Hemogenic endothelium
Human embryonic stem cells
Human hematopoiesis
Humans
Pluripotency
Pluripotent Stem Cells - cytology
Pluripotent Stem Cells - metabolism
Progenitor cells
Ribonucleic acid
RNA
Single cell RNA‐seq
Stem cells
Tissue inhibitor of metalloproteinase 3
Transcription
Human embryonic stem cells
Human hematopoiesis
Single cell RNA-seq
Hemogenic endothelium
Online AccessGet full text
ISSN1066-5099
1549-4918
1549-4918
DOI10.1002/stem.2739

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Abstract Endothelial‐to‐hematopoietic transition (EHT) is an important stage in definitive hematopoietic development. However, the genetic mechanisms underlying human EHT remain poorly characterized. We performed single cell RNA‐seq using 55 hemogenic endothelial cells (HECs: CD31+CD144+CD41–CD43–CD45–CD73–RUNX1c+), 47 vascular endothelial cells without hematopoietic potential (non‐HE: CD31+CD144+CD41–CD43–CD45–CD73–RUNX1c–), and 35 hematopoietic progenitor cells (HPCs: CD34+CD43+RUNX1c+) derived from human embryonic stem cells (hESCs). HE and HP were enriched in genes implicated in hemogenic endothelial transcriptional networks, such as ERG, GATA2, and FLI. We found transcriptional overlap between individual HECs and HPCs; however, these populations were distinct from non‐HE. Further analysis revealed novel biomarkers for human HEC/HPCs, including TIMP3, ESAM, RHOJ, and DLL4. Collectively, we demonstrate that hESC‐derived HE and HP share a common developmental pathway, while non‐HE are more heterogeneous and transcriptionally distinct. Our findings provide a novel strategy to test new genetic targets and optimize the production of definitive hematopoietic cells from human pluripotent stem cells. Stem Cells 2018;36:206–217 Using a RUNX1c reporter hESC line, we derived functional hemogenic endothelium, non‐hemogenic endothelium, and early hematopoietic progenitor cells from embryoid bodies. With single‐cell RNA‐sequencing, we characterized the genetic signatures distinct to each population and further identified novel genetic targets that may be used to optimize production of definitive human hematopoietic cells from pluripotent stem cells.
AbstractList Endothelial-to-hematopoietic transition (EHT) is an important stage in definitive hematopoietic development. However, the genetic mechanisms underlying human EHT remain poorly characterized. We performed single cell RNA-seq using 55 hemogenic endothelial cells (HECs: CD31+ CD144+ CD41- CD43- CD45- CD73- RUNX1c+ ), 47 vascular endothelial cells without hematopoietic potential (non-HE: CD31+ CD144+ CD41- CD43- CD45- CD73- RUNX1c- ), and 35 hematopoietic progenitor cells (HPCs: CD34+ CD43+ RUNX1c+ ) derived from human embryonic stem cells (hESCs). HE and HP were enriched in genes implicated in hemogenic endothelial transcriptional networks, such as ERG, GATA2, and FLI. We found transcriptional overlap between individual HECs and HPCs; however, these populations were distinct from non-HE. Further analysis revealed novel biomarkers for human HEC/HPCs, including TIMP3, ESAM, RHOJ, and DLL4. Collectively, we demonstrate that hESC-derived HE and HP share a common developmental pathway, while non-HE are more heterogeneous and transcriptionally distinct. Our findings provide a novel strategy to test new genetic targets and optimize the production of definitive hematopoietic cells from human pluripotent stem cells. Stem Cells 2018;36:206-217.Endothelial-to-hematopoietic transition (EHT) is an important stage in definitive hematopoietic development. However, the genetic mechanisms underlying human EHT remain poorly characterized. We performed single cell RNA-seq using 55 hemogenic endothelial cells (HECs: CD31+ CD144+ CD41- CD43- CD45- CD73- RUNX1c+ ), 47 vascular endothelial cells without hematopoietic potential (non-HE: CD31+ CD144+ CD41- CD43- CD45- CD73- RUNX1c- ), and 35 hematopoietic progenitor cells (HPCs: CD34+ CD43+ RUNX1c+ ) derived from human embryonic stem cells (hESCs). HE and HP were enriched in genes implicated in hemogenic endothelial transcriptional networks, such as ERG, GATA2, and FLI. We found transcriptional overlap between individual HECs and HPCs; however, these populations were distinct from non-HE. Further analysis revealed novel biomarkers for human HEC/HPCs, including TIMP3, ESAM, RHOJ, and DLL4. Collectively, we demonstrate that hESC-derived HE and HP share a common developmental pathway, while non-HE are more heterogeneous and transcriptionally distinct. Our findings provide a novel strategy to test new genetic targets and optimize the production of definitive hematopoietic cells from human pluripotent stem cells. Stem Cells 2018;36:206-217.
Endothelial‐to‐hematopoietic transition (EHT) is an important stage in definitive hematopoietic development. However, the genetic mechanisms underlying human EHT remain poorly characterized. We performed single cell RNA‐seq using 55 hemogenic endothelial cells (HECs: CD31+CD144+CD41–CD43–CD45–CD73–RUNX1c+), 47 vascular endothelial cells without hematopoietic potential (non‐HE: CD31+CD144+CD41–CD43–CD45–CD73–RUNX1c–), and 35 hematopoietic progenitor cells (HPCs: CD34+CD43+RUNX1c+) derived from human embryonic stem cells (hESCs). HE and HP were enriched in genes implicated in hemogenic endothelial transcriptional networks, such as ERG, GATA2, and FLI. We found transcriptional overlap between individual HECs and HPCs; however, these populations were distinct from non‐HE. Further analysis revealed novel biomarkers for human HEC/HPCs, including TIMP3, ESAM, RHOJ, and DLL4. Collectively, we demonstrate that hESC‐derived HE and HP share a common developmental pathway, while non‐HE are more heterogeneous and transcriptionally distinct. Our findings provide a novel strategy to test new genetic targets and optimize the production of definitive hematopoietic cells from human pluripotent stem cells. Stem Cells 2018;36:206–217 Using a RUNX1c reporter hESC line, we derived functional hemogenic endothelium, non‐hemogenic endothelium, and early hematopoietic progenitor cells from embryoid bodies. With single‐cell RNA‐sequencing, we characterized the genetic signatures distinct to each population and further identified novel genetic targets that may be used to optimize production of definitive human hematopoietic cells from pluripotent stem cells.
Endothelial‐to‐hematopoietic transition (EHT) is an important stage in definitive hematopoietic development. However, the genetic mechanisms underlying human EHT remain poorly characterized. We performed single cell RNA‐seq using 55 hemogenic endothelial cells (HECs: CD31+CD144+CD41–CD43–CD45–CD73–RUNX1c+), 47 vascular endothelial cells without hematopoietic potential (non‐HE: CD31+CD144+CD41–CD43–CD45–CD73–RUNX1c–), and 35 hematopoietic progenitor cells (HPCs: CD34+CD43+RUNX1c+) derived from human embryonic stem cells (hESCs). HE and HP were enriched in genes implicated in hemogenic endothelial transcriptional networks, such as ERG, GATA2, and FLI. We found transcriptional overlap between individual HECs and HPCs; however, these populations were distinct from non‐HE. Further analysis revealed novel biomarkers for human HEC/HPCs, including TIMP3, ESAM, RHOJ, and DLL4. Collectively, we demonstrate that hESC‐derived HE and HP share a common developmental pathway, while non‐HE are more heterogeneous and transcriptionally distinct. Our findings provide a novel strategy to test new genetic targets and optimize the production of definitive hematopoietic cells from human pluripotent stem cells. Stem Cells 2018;36:206–217
Endothelial-to-hematopoietic transition (EHT) is an important stage in definitive hematopoietic development. However, the genetic mechanisms underlying human EHT remain poorly characterized. We performed single cell RNA-seq using 55 hemogenic endothelial cells (HECs: CD31 CD144 CD41 CD43 CD45 CD73 RUNX1c ), 47 vascular endothelial cells without hematopoietic potential (non-HE: CD31 CD144 CD41 CD43 CD45 CD73 RUNX1c ), and 35 hematopoietic progenitor cells (HPCs: CD34 CD43 RUNX1c ) derived from human embryonic stem cells (hESCs). HE and HP were enriched in genes implicated in hemogenic endothelial transcriptional networks, such as ERG, GATA2, and FLI. We found transcriptional overlap between individual HECs and HPCs; however, these populations were distinct from non-HE. Further analysis revealed novel biomarkers for human HEC/HPCs, including TIMP3, ESAM, RHOJ, and DLL4. Collectively, we demonstrate that hESC-derived HE and HP share a common developmental pathway, while non-HE are more heterogeneous and transcriptionally distinct. Our findings provide a novel strategy to test new genetic targets and optimize the production of definitive hematopoietic cells from human pluripotent stem cells. Stem Cells 2018;36:206-217.
Endothelial-to-hematopoietic transition (EHT) is an important stage in definitive hematopoietic development. However, the genetic mechanisms underlying human EHT remain poorly characterized. We performed single cell RNA-seq using 55 hemogenic endothelial cells (HECs: CD31+CD144+CD41–CD43–CD45–CD73–RUNX1c+), 47 vascular endothelial cells without hematopoietic potential (non-HE: CD31+CD144+CD41–CD43–CD45–CD73–RUNX1c–), and 35 hematopoietic progenitor cells (HPCs: CD34+CD43+RUNX1c+) derived from human embryonic stem cells (hESCs). HE and HP were enriched in genes implicated in hemogenic endothelial transcriptional networks, such as ERG, GATA2, and FLI. We found transcriptional overlap between individual HECs and HPCs; however, these populations were distinct from non-HE. Further analysis revealed novel biomarkers for human HEC/HPCs, including TIMP3, ESAM, RHOJ, and DLL4. Collectively, we demonstrate that hESC-derived HE and HP share a common developmental pathway, while non-HE are more heterogeneous and transcriptionally distinct. Our findings provide a novel strategy to test new genetic targets and optimize the production of definitive hematopoietic cells from human pluripotent stem cells.
Author Angelos, Mathew G.
Abrahante, Juan E.
Blum, Robert H.
Kaufman, Dan S.
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  surname: Kaufman
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Human hematopoiesis
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Hemogenic endothelium
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Snippet Endothelial‐to‐hematopoietic transition (EHT) is an important stage in definitive hematopoietic development. However, the genetic mechanisms underlying human...
Endothelial-to-hematopoietic transition (EHT) is an important stage in definitive hematopoietic development. However, the genetic mechanisms underlying human...
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SubjectTerms Antigens, Surface - metabolism
Biomarkers
CD34 antigen
CD43 antigen
CD45 antigen
CD73 antigen
Cell Differentiation - genetics
Cell Differentiation - physiology
Cells (biology)
Cells, Cultured
Computational Biology
Developmental stages
Embryo cells
Embryonic Stem Cells - cytology
Embryonic Stem Cells - metabolism
Embryos
Endothelial cells
Endothelium
Flow Cytometry
Hemangioblasts - cytology
Hemangioblasts - metabolism
Hematopoiesis - genetics
Hematopoiesis - physiology
Hematopoietic stem cells
Hematopoietic Stem Cells - cytology
Hematopoietic Stem Cells - metabolism
Hemogenic endothelium
Human embryonic stem cells
Human hematopoiesis
Humans
Pluripotency
Pluripotent Stem Cells - cytology
Pluripotent Stem Cells - metabolism
Progenitor cells
Ribonucleic acid
RNA
Single cell RNA‐seq
Stem cells
Tissue inhibitor of metalloproteinase 3
Transcription
Title Single Cell Resolution of Human Hematoendothelial Cells Defines Transcriptional Signatures of Hemogenic Endothelium
URI https://onlinelibrary.wiley.com/doi/abs/10.1002%2Fstem.2739
https://www.ncbi.nlm.nih.gov/pubmed/29139170
https://www.proquest.com/docview/1990847360
https://www.proquest.com/docview/1964698028
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