Human yolk sac-like haematopoiesis generates RUNX1-, GFI1- and/or GFI1B- dependent blood and SOX17-positive endothelium

Freya F Bruveris; Elizabeth S Ng; Ana Rita Leitoguinho; Ali Motazedian; Katerina Vlahos; Koula Sourris; Robyn Mayberry; Penelope McDonald; Lisa Azzola; Nadia M Davidson; Alicia Oshlack; Edouard G Stanley; Andrew G Elefanty

doi:10.1242/dev.193037

Human yolk sac-like haematopoiesis generates RUNX1-, GFI1- and/or GFI1B- dependent blood and SOX17-positive endothelium

Development. 2020 Oct 29;147(20):dev193037. doi: 10.1242/dev.193037.

Authors

Freya F Bruveris^{1

2}, Elizabeth S Ng¹, Ana Rita Leitoguinho^{1

2}, Ali Motazedian^{1

2}, Katerina Vlahos¹, Koula Sourris¹, Robyn Mayberry¹, Penelope McDonald¹, Lisa Azzola¹, Nadia M Davidson^{1

3}, Alicia Oshlack^{1

3}, Edouard G Stanley^{1

2

4}, Andrew G Elefanty^{5

2

4}

Affiliations

¹ Murdoch Children's Research Institute, The Royal Children's Hospital, Flemington Road, Parkville, Victoria 3052, Australia.
² Department of Paediatrics, Faculty of Medicine, Dentistry and Health Sciences, University of Melbourne, Parkville, Victoria 3052, Australia.
³ School of BioSciences, University of Melbourne, Parkville, Victoria 3052, Australia.
⁴ Department of Anatomy and Developmental Biology, Monash University, Clayton, Victoria 3800, Australia.
⁵ Murdoch Children's Research Institute, The Royal Children's Hospital, Flemington Road, Parkville, Victoria 3052, Australia andrew.elefanty@mcri.edu.au.

Abstract

The genetic regulatory network controlling early fate choices during human blood cell development are not well understood. We used human pluripotent stem cell reporter lines to track the development of endothelial and haematopoietic populations in an in vitro model of human yolk-sac development. We identified SOX17^-CD34⁺CD43^- endothelial cells at day 2 of blast colony development, as a haemangioblast-like branch point from which SOX17^-CD34⁺CD43⁺ blood cells and SOX17⁺CD34⁺CD43^- endothelium subsequently arose. Most human blood cell development was dependent on RUNX1. Deletion of RUNX1 only permitted a single wave of yolk sac-like primitive erythropoiesis, but no yolk sac myelopoiesis or aorta-gonad-mesonephros (AGM)-like haematopoiesis. Blocking GFI1 and/or GFI1B activity with a small molecule inhibitor abrogated all blood cell development, even in cell lines with an intact RUNX1 gene. Together, our data define the hierarchical requirements for RUNX1, GFI1 and/or GFI1B during early human haematopoiesis arising from a yolk sac-like SOX17-negative haemogenic endothelial intermediate.

Keywords: GFI1/1B; Human pluripotent stem cells; RUNX1; Yolk sac haematopoiesis.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Blood Cells / cytology
Blood Cells / metabolism*
Cell Differentiation
Cell Lineage
Core Binding Factor Alpha 2 Subunit / metabolism*
DNA-Binding Proteins / metabolism*
Endothelium / metabolism*
Erythroid Cells / cytology
Erythroid Cells / metabolism
Hematopoiesis*
Histone Demethylases / antagonists & inhibitors
Histone Demethylases / metabolism
Humans
Models, Biological
Proto-Oncogene Proteins / metabolism*
Repressor Proteins / metabolism*
SOXF Transcription Factors / metabolism*
Transcription Factors / metabolism*
Transcription, Genetic
Yolk Sac / metabolism*

Substances

Core Binding Factor Alpha 2 Subunit
DNA-Binding Proteins
GFI1 protein, human
GFI1B protein, human
Proto-Oncogene Proteins
Repressor Proteins
SOX17 protein, human
SOXF Transcription Factors
Transcription Factors
Histone Demethylases
KDM1A protein, human