Runx1 is required for the endothelial to haematopoietic cell transition but not thereafter

Abstract

Haematopoietic stem cells (HSCs) are the founder cells of the adult haematopoietic system, and thus knowledge of the molecular program directing their generation during development is important for regenerative haematopoietic strategies. Runx1 is a pivotal transcription factor required for HSC generation in the vascular regions of the mouse conceptus—the aorta, vitelline and umbilical arteries, yolk sac and placenta1,2. It is thought that HSCs emerge from vascular endothelial cells through the formation of intra-arterial clusters3 and that Runx1 functions during the transition from ‘haemogenic endothelium’ to HSCs4,5. Here we show by conditional deletion that Runx1 activity in vascular-endothelial-cadherin-positive endothelial cells is indeed essential for intra-arterial cluster, haematopoietic progenitor and HSC formation in mice. In contrast, Runx1 is not required in cells expressing Vav1, one of the first pan-haematopoietic genes expressed in HSCs. Collectively these data show that Runx1 function is essential in endothelial cells for haematopoietic progenitor and HSC formation from the vasculature, but its requirement ends once or before Vav is expressed.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Figure 1: VEC-Cre excision marks endothelium and blood.
Figure 2: Runx1 is required in VEC + cells for haematopoietic progenitor formation.
Figure 3: Runx1 is required in VEC + cells for HSC emergence.
Figure 4: Runx1 is not required in Vav + cells for CFU-C or HSC emergence.

References

  1. 1

    Cai, Z. et al. Haploinsufficiency of AML1/CBFA2 affects the embryonic generation of mouse hematopoietic stem cells. Immunity 13, 423–431 (2000)

    CAS  Article  Google Scholar 

  2. 2

    North, T. E. et al. Runx1 expression marks long-term repopulating hematopoietic stem cells in the midgestation mouse embryo. Immunity 16, 661–672 (2002)

    CAS  Article  Google Scholar 

  3. 3

    Jaffredo, T., Gautier, R., Eichmann, A. & Dieterlen-Lièvre, F. Intraaortic hemopoietic cells are derived from endothelial cells during ontogeny. Development 125, 4575–4583 (1998)

    CAS  PubMed  Google Scholar 

  4. 4

    North, T. E. et al. Cbfa2 is required for the formation of intra-aortic hematopoietic clusters. Development 126, 2563–2575 (1999)

    CAS  PubMed  Google Scholar 

  5. 5

    Yokomizo, T. et al. Requirement of Runx1/AML1/PEBP2αB for the generation of haematopoietic cells from endothelial cells. Genes Cells 6, 13–23 (2001)

    CAS  Article  Google Scholar 

  6. 6

    Samokhvalov, I. M., Samokhvalova, N. I. & Nishikawa, S. I. Cell tracing shows the contribution of the yolk sac to adult haematopoiesis. Nature 446, 1056–1061 (2007)

    ADS  CAS  Article  Google Scholar 

  7. 7

    Zeigler, B. M. et al. The allantois and chorion, when isolated before circulation or chorio-allantoic fusion, have hematopoietic potential. Development 133, 4183–4192 (2006)

    CAS  Article  Google Scholar 

  8. 8

    Rhodes, K. E. et al. The emergence of hematopoietic stem cells is initiated in the placental vasculature in the absence of circulation. Cell Stem Cell 2, 252–263 (2008)

    CAS  Article  Google Scholar 

  9. 9

    Lampugnani, M. G. et al. The molecular organization of endothelial cell to cell junctions: differential association of plakoglobin, β-catenin, and α-catenin with vascular endothelial cadherin (VE-cadherin). J. Cell Biol. 129, 203–217 (1995)

    CAS  Article  Google Scholar 

  10. 10

    Breier, G. et al. Molecular cloning and expression of murine vascular endothelial-cadherin in early stage development of cardiovascular system. Blood 87, 630–641 (1996)

    CAS  Article  Google Scholar 

  11. 11

    Drake, C. J. & Fleming, P. A. Vasculogenesis in the day 6.5 to 9.5 mouse embryo. Blood 95, 1671–1679 (2000)

    CAS  Article  Google Scholar 

  12. 12

    Yokomizo, T. et al. Characterization of GATA-1+ hemangioblastic cells in the mouse embryo. EMBO J. 26, 184–196 (2007)

    CAS  Article  Google Scholar 

  13. 13

    Kinder, S. J. et al. The orderly allocation of mesodermal cells to the extraembryonic structures and the anteroposterior axis during gastrulation of the mouse embryo. Development 126, 4691–4701 (1999)

    CAS  PubMed  Google Scholar 

  14. 14

    Lawson, K. A., Meneses, J. J. & Pedersen, R. A. Clonal analysis of epiblast fate during germ layer formation in the mouse embryo. Development 113, 891–911 (1991)

    CAS  PubMed  Google Scholar 

  15. 15

    Downs, K. M., Hellman, E. R., McHugh, J., Barrickman, K. & Inman, K. E. Investigation into a role for the primitive streak in development of the murine allantois. Development 131, 37–55 (2004)

    CAS  Article  Google Scholar 

  16. 16

    de Bruijn, M. et al. Hematopoietic stem cells localize to the endothelial cell layer in the midgestation mouse aorta. Immunity 16, 673–683 (2002)

    CAS  Article  Google Scholar 

  17. 17

    Okuda, T., van Deursen, J., Hiebert, S. W., Grosveld, G. & Downing, J. R. AML1, the target of multiple chromosomal translocations in human leukemia, is essential for normal fetal liver hematopoiesis. Cell 84, 321–330 (1996)

    CAS  Article  Google Scholar 

  18. 18

    Wang, Q. et al. Disruption of the Cbfa2 gene causes necrosis and hemorrhaging in the central nervous system and blocks definitive hematopoiesis. Proc. Natl Acad. Sci. USA 93, 3444–3449 (1996)

    ADS  CAS  Article  Google Scholar 

  19. 19

    de Bruijn, M. F. T. R., Speck, N. A., Peeters, M. C. E. & Dzierzak, E. Definitive hematopoietic stem cells first emerge from the major arterial regions of the mouse embryo. EMBO J. 19, 2465–2474 (2000)

    CAS  Article  Google Scholar 

  20. 20

    Ogilvy, S. et al. Promoter elements of vav drive transgene expression in vivo throughout the hematopoietic compartment. Blood 94, 1855–1863 (1999)

    CAS  Article  Google Scholar 

  21. 21

    Stadtfeld, M. & Graf, T. Assessing the role of hematopoietic plasticity for endothelial and hepatocyte development by non-invasive lineage tracing. Development 132, 203–213 (2005)

    CAS  Article  Google Scholar 

  22. 22

    Okada, H. et al. AML1-/- embryos do not express certain hematopoiesis-related gene transcripts including those of the PU.1 gene. Oncogene 17, 2287–2293 (1998)

    CAS  Article  Google Scholar 

  23. 23

    Kim, I., Yilmaz, O. H. & Morrison, S. J. CD144 (VE-cadherin) is transiently expressed by fetal liver hematopoietic stem cells. Blood 106, 903–905 (2005)

    CAS  Article  Google Scholar 

  24. 24

    Taoudi, S. et al. Progressive divergence of definitive haematopoietic stem cells from the endothelial compartment does not depend on contact with the foetal liver. Development 132, 4179–4191 (2005)

    CAS  Article  Google Scholar 

  25. 25

    Ichikawa, M. et al. AML-1 is required for megakaryocytic maturation and lymphocytic differentiation, but not for maintenance of hematopoietic stem cells in adult hematopoiesis. Nature Med. 10, 299–304 (2004)

    CAS  Article  Google Scholar 

  26. 26

    Growney, J. D. et al. Loss of Runx1 perturbs adult hematopoiesis and is associated with a myeloproliferative phenotype. Blood 106, 494–504 (2005)

    CAS  Article  Google Scholar 

  27. 27

    Monvoisin, A. et al. VE-cadherin-CreERT2 transgenic mouse: a model for inducible recombination in the endothelium. Dev. Dyn. 235, 3413–3422 (2006)

    CAS  Article  Google Scholar 

  28. 28

    Bertrand, J. Y. et al. Characterization of purified intraembryonic hematopoietic stem cells as a tool to define their site of origin. Proc. Natl Acad. Sci. USA 102, 134–139 (2005)

    ADS  CAS  Article  Google Scholar 

  29. 29

    Chung, J. H., Whiteley, M. & Felsenfeld, G. A 5′ element of the chicken beta-globin domain serves as an insulator in human erythroid cells and protects against position effect in Drosophila . Cell 74, 505–514 (1993)

    CAS  Article  Google Scholar 

  30. 30

    Inman, K. E. & Downs, K. M. Brachyury is required for elongation and vasculogenesis in the murine allantois. Development 133, 2947–2959 (2006)

    CAS  Article  Google Scholar 

  31. 31

    Gory, B. et al. The vascular endothelial-cadherin promoter directs endothelial-specific expression in transgenic mice. Blood 93, 184–192 (1999)

    CAS  Article  Google Scholar 

Download references

Acknowledgements

The authors thank G. Ward for his assistance with flow, K. Downs for technical advice, T. Graf for the Vav-Cre mice, and P. Huber for the Cdh5 sequences. This work was supported by R01HL091724 (N.A.S.), R01DK54077 (E.D.) and T32 AI-07519 (B.M.Z.). Core services were supported in part by the Norris Cotton Cancer Center (NIH CA23108) and the Abramson Family Cancer Research Institute.

Author Contributions T.Y. performed the experiments in Fig. 3d. B.M.Z. performed the experiments in Fig. 1b, d. M.J.C. performed all the remaining experiments. E.D. participated in the interpretation of the experiments and writing the manuscript. N.A.S. participated in the design and interpretation of the experiments, wrote the manuscript and made the figures.

Author information

Affiliations

Authors

Corresponding author

Correspondence to Nancy A. Speck.

Supplementary information

Supplementary Information

This file contains Supplementary Figures 1-2 with Legends and Supplementary Tables 1-5 (PDF 754 kb)

PowerPoint slides

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Chen, M., Yokomizo, T., Zeigler, B. et al. Runx1 is required for the endothelial to haematopoietic cell transition but not thereafter. Nature 457, 887–891 (2009). https://doi.org/10.1038/nature07619

Download citation

Further reading

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing