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HoxA3 is an apical regulator of haemogenic endothelium

Nature Cell Biology volume 13, pages 72–78 (2011)Cite this article

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Abstract

During development, haemogenesis occurs invariably at sites of vasculogenesis. Between embryonic day (E) 9.5 and E10.5 in mice, endothelial cells in the caudal part of the dorsal aorta generate haematopoietic stem cells1,2 and are referred to as haemogenic endothelium3,4,5,6,7,8. The mechanisms by which haematopoiesis is restricted to this domain, and how the morphological transformation from endothelial to haematopoietic is controlled are unknown. We show here that HoxA3, a gene uniquely expressed in the embryonic but not yolk sac vasculature, restrains haematopoietic differentiation of the earliest endothelial progenitors, and induces reversion of the earliest haematopoietic progenitors into CD41-negative endothelial cells. This reversible modulation of endothelial–haematopoietic state is accomplished by targeting key haematopoietic transcription factors for downregulation, including Runx1, Gata1, Gfi1B, Ikaros, and PU.1. Through loss-of-function, and gain-of-function epistasis experiments, and the identification of antipodally regulated targets, we show that among these factors, Runx1 is uniquely able to erase the endothelial program set up by HoxA3. These results suggest both why a frank endothelium does not precede haematopoiesis in the yolk sac, and why haematopoietic stem cell generation requires Runx1 expression only in endothelial cells.

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Figure 1: Reciprocal expression of HoxA3 and Runx1 in embryonic endothelium.
Figure 2: HoxA3 expression in early mesoderm and committed haemogenic endothelium restrains haematopoiesis.
Figure 3: Global expression changes on HoxA3 induction.
Figure 4: Global expression changes on reversion of HoxA3 by Runx1 or Gata1.
Figure 5: Regulation of intraembryonic haematopoiesis by HoxA3 and Runx1.

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Acknowledgements

We thank the Bob and Jean Smith Foundation for their generous support. This work was supported by the NIH grant 1R01HL081186-01 and the March of Dimes grant 5-FY2006-272. We thank Nardina Nash for genotyping and animal husbandry.

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Authors and Affiliations

  1. Lillehei Heart Institute and Department of Pediatrics, University of Minnesota, 312 Church St. SE, Minneapolis, 55455, MN, USA

    Michelina Iacovino, Istvan Szatmari, Lynn Hartweck, Danielle Rux & Michael Kyba

  2. Department of Molecular Biology, UT Southwestern Medical Center, 5323 Harry Hines Blvd, Dallas, 75390-9148, TX, USA

    Diana Chong, Arianna Caprioli & Ondine Cleaver

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  1. Michelina Iacovino
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Contributions

M.I., O.C. and M.K. designed the experiments, and wrote the manuscript; M.I. performed the experiments; D.C. and A.C. performed in situ hybridization studies; I.S. performed microarray studies; L.H. and D.R. performed chromatin IP studies.

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Correspondence to Michael Kyba.

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Iacovino, M., Chong, D., Szatmari, I. et al. HoxA3 is an apical regulator of haemogenic endothelium. Nat Cell Biol 13, 72–78 (2011). https://doi.org/10.1038/ncb2137

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