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A somitic Wnt16/Notch pathway specifies haematopoietic stem cells

Nature volume 474, pages 220–224 (2011)Cite this article

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Abstract

Haematopoietic stem cells (HSCs) are a self-renewing population of cells that continuously replenish all blood and immune cells during the lifetime of an individual1,2. HSCs are used clinically to treat a wide array of diseases, including acute leukaemias and congenital blood disorders, but obtaining suitable numbers of cells and finding immune-compatible donors remain serious problems. These difficulties have led to an interest in the conversion of embryonic stem cells or induced pluripotent stem cells into HSCs, which is not possible using current methodologies. To accomplish this goal, it is critical to understand the native mechanisms involved in the specification of HSCs during embryonic development. Here we demonstrate in zebrafish that Wnt16 controls a novel genetic regulatory network required for HSC specification. Non-canonical signalling by Wnt16 is required for somitic expression of the Notch ligands deltaC (dlc) and deltaD (dld), and these ligands are, in turn, required for the establishment of definitive haematopoiesis. Notch signalling downstream of Dlc and Dld is earlier than, and distinct from, known cell-autonomous requirements for Notch, strongly suggesting that novel Notch-dependent relay signal(s) induce the first HSCs in parallel to other established pathways. Our results demonstrate that somite-specific gene expression is required for the production of haemogenic endothelium.

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Figure 1: Wnt16 is required for the specification of HSCs.
Figure 2: The wnt16 loss-of-function phenotype is specific.
Figure 3: Wnt16 acts upstream of Notch ligands Dlc and Dld.
Figure 4: Non-cell-autonomous requirement for Notch in HSC specification.

References

  1. Gering, M. & Patient, R. Notch signalling and haematopoietic stem cell formation during embryogenesis. J. Cell. Physiol. 222, 11–16 (2010)

    Article  CAS  Google Scholar 

  2. Staal, F. J. & Luis, T. C. Wnt signaling in hematopoiesis: crucial factors for self-renewal, proliferation, and cell fate decisions. J. Cell. Biochem. 109, 844–849 (2010)

    CAS  PubMed  Google Scholar 

  3. Angers, S. & Moon, R. T. Proximal events in Wnt signal transduction. Nature Rev. Mol. Cell Biol. 10, 468–477 (2009)

    Article  CAS  Google Scholar 

  4. Takada, S. et al. Wnt-3a regulates somite and tailbud formation in the mouse embryo. Genes Dev. 8, 174–189 (1994)

    Article  CAS  Google Scholar 

  5. Goessling, W. et al. Genetic interaction of PGE2 and Wnt signaling regulates developmental specification of stem cells and regeneration. Cell 136, 1136–1147 (2009)

    Article  CAS  Google Scholar 

  6. McWhirter, J. R. et al. Oncogenic homeodomain transcription factor E2A-Pbx1 activates a novel WNT gene in pre-B acute lymphoblastoid leukemia. Proc. Natl Acad. Sci. USA 96, 11464–11469 (1999)

    Article  CAS  ADS  Google Scholar 

  7. Bertrand, J. Y. et al. Haematopoietic stem cells derive directly from aortic endothelium during development. Nature 464, 108–111 (2010)

    Article  CAS  ADS  Google Scholar 

  8. Kissa, K. & Herbomel, P. Blood stem cells emerge from aortic endothelium by a novel type of cell transition. Nature 464, 112–115 (2010)

    Article  CAS  ADS  Google Scholar 

  9. Boisset, J. C. et al. In vivo imaging of haematopoietic cells emerging from the mouse aortic endothelium. Nature 464, 116–120 (2010)

    Article  CAS  ADS  Google Scholar 

  10. Bertrand, J. Y. et al. Definitive hematopoiesis initiates through a committed erythromyeloid precursor in the zebrafish embryo. Development 134, 4147–4156 (2007)

    Article  CAS  Google Scholar 

  11. Kissa, K. et al. Live imaging of emerging hematopoietic stem cells and early thymus colonization. Blood 111, 1147–1156 (2008)

    Article  CAS  Google Scholar 

  12. Lin, H. F. et al. Analysis of thrombocyte development in CD41-GFP transgenic zebrafish. Blood 106, 3803–3810 (2005)

    Article  CAS  Google Scholar 

  13. Yokota, T. et al. Tracing the first waves of lymphopoiesis in mice. Development 133, 2041–2051 (2006)

    Article  CAS  Google Scholar 

  14. Clements, W. K., Ong, K. G. & Traver, D. Zebrafish wnt3 is expressed in developing neural tissue. Dev. Dyn. 238, 1788–1795 (2009)

    Article  CAS  Google Scholar 

  15. Nygren, M. K. et al. β-catenin is involved in N-cadherin-dependent adhesion, but not in canonical Wnt signaling in E2A–PBX1-positive B acute lymphoblastic leukemia cells. Exp. Hematol. 37, 225–233 (2009)

    Article  CAS  Google Scholar 

  16. Teh, M. T. et al. Role for WNT16B in human epidermal keratinocyte proliferation and differentiation. J. Cell Sci. 120, 330–339 (2007)

    Article  CAS  Google Scholar 

  17. Cirone, P. et al. A role for planar cell polarity signaling in angiogenesis. Angiogenesis 11, 347–360 (2008)

    Article  Google Scholar 

  18. Julich, D. et al. beamter/deltaC and the role of Notch ligands in the zebrafish somite segmentation, hindbrain neurogenesis and hypochord differentiation. Dev. Biol. 286, 391–404 (2005)

    Article  Google Scholar 

  19. Holley, S. A., Julich, D., Rauch, G. J., Geisler, R. & Nusslein-Volhard, C. her1 and the notch pathway function within the oscillator mechanism that regulates zebrafish somitogenesis. Development 129, 1175–1183 (2002)

    CAS  PubMed  Google Scholar 

  20. Burns, C. E., Traver, D., Mayhall, E., Shepard, J. L. & Zon, L. I. Hematopoietic stem cell fate is established by the Notch-Runx pathway. Genes Dev. 19, 2331–2342 (2005)

    Article  CAS  Google Scholar 

  21. Hadland, B. K. et al. A requirement for Notch1 distinguishes 2 phases of definitive hematopoiesis during development. Blood 104, 3097–3105 (2004)

    Article  CAS  Google Scholar 

  22. Kumano, K. et al. Notch1 but not Notch2 is essential for generating hematopoietic stem cells from endothelial cells. Immunity 18, 699–711 (2003)

    Article  CAS  Google Scholar 

  23. Robert-Moreno, A., Espinosa, L., de la Pompa, J. L. & Bigas, A. RBPjκ-dependent Notch function regulates Gata2 and is essential for the formation of intra-embryonic hematopoietic cells. Development 132, 1117–1126 (2005)

    Article  CAS  Google Scholar 

  24. Robert-Moreno, A. et al. Impaired embryonic haematopoiesis yet normal arterial development in the absence of the Notch ligand Jagged1. EMBO J. 27, 1886–1895 (2008)

    Article  CAS  Google Scholar 

  25. Scheer, N. & Campos-Ortega, J. A. Use of the Gal4-UAS technique for targeted gene expression in the zebrafish. Mech. Dev. 80, 153–158 (1999)

    Article  CAS  Google Scholar 

  26. Scheer, N., Groth, A., Hans, S. & Campos-Ortega, J. A. An instructive function for Notch in promoting gliogenesis in the zebrafish retina. Development 128, 1099–1107 (2001)

    CAS  PubMed  Google Scholar 

  27. Ren, X., Gomez, G. A., Zhang, B. & Lin, S. Scl isoforms act downstream of etsrp to specify angioblasts and definitive hematopoietic stem cells. Blood 115, 5338–5346 (2010)

    Article  CAS  Google Scholar 

  28. Kemp, C., Willems, E., Abdo, S., Lambiv, L. & Leyns, L. Expression of all Wnt genes and their secreted antagonists during mouse blastocyst and postimplantation development. Dev. Dyn. 233, 1064–1075 (2005)

    Article  CAS  Google Scholar 

  29. Corrigan, P. M., Dobbin, E., Freeburn, R. W. & Wheadon, H. Patterns of Wnt/Fzd/LRP gene expression during embryonic hematopoiesis. Stem Cells Dev. 18, 759–772 (2009)

    Article  CAS  Google Scholar 

  30. Westerfield, M. The Zebrafish Book. A Guide for the Laboratory Use of Zebrafish (Danio Rerio) (Univ. Oregon Press, 2004)

    Google Scholar 

  31. Parsons, M. J. et al. Notch-responsive cells initiate the secondary transition in larval zebrafish pancreas. Mech. Dev. 126, 898–912 (2009)

    Article  CAS  Google Scholar 

  32. Turner, D. L. & Weintraub, H. Expression of achaete-scute homolog 3 in Xenopus embryos converts ectodermal cells to a neural fate. Genes Dev. 8, 1434–1447 (1994)

    Article  CAS  Google Scholar 

  33. Clements, W. K. & Kimelman, D. LZIC regulates neuronal survival during zebrafish development. Dev. Biol. 283, 322–334 (2005)

    Article  CAS  Google Scholar 

  34. Lele, Z., Bakkers, J. & Hammerschmidt, M. Morpholino phenocopies of the swirl, snailhouse, somitabun, minifin, silberblick, and pipetail mutations. Genesis 30, 190–194 (2001)

    Article  CAS  Google Scholar 

  35. Nüsslein-Volhard, C. & Dahm, R. Zebrafish (Oxford Univ. Press, 2002)

    Google Scholar 

  36. Bertrand, J. Y., Kim, A. D., Teng, S. & Traver, D. CD41+ cmyb+ precursors colonize the zebrafish pronephros by a novel migration route to initiate adult hematopoiesis. Development 135, 1853–1862 (2008)

    Article  CAS  Google Scholar 

Download references

Acknowledgements

The authors wish to thank L. Zon, K. Poss, D. Kimelman, M. Lardelli, B. Appel, C. Burns, J. Campos-Ortega, D. Ransom, N. Trede, J. Lewis, M. Pack, S. Holley, C. Moens, B. Paw, R. Karlström and J. Waxman for probe constructs. L. Zon, R. Dorsky, S. Lin and S. Holley provided transgenic and mutant zebrafish lines. C. Weaver, K. Willert, K. J. P. Griffin, J. Bertrand, D. Stachura and Y. Lee provided critical evaluation of the manuscript. This research was funded by an AHA Postdoctoral Fellowship 0725086Y to W.K.C., an AHA Predoctoral Founders Affiliate Fellowship 0815732D to J.C.M., NIH R01-HL093467 to N.L. and NIH R01-DK074482, CIRM New Investigator Award, and March of Dimes 6-FY09-508 to D.T.

Author information

Authors and Affiliations

  1. Department of Cellular and Molecular Medicine and Section of Cell and Developmental Biology, University of California at San Diego, 9500 Gilman Drive, La Jolla, California 92093-0380, USA,

    Wilson K. Clements, Albert D. Kim, Karen G. Ong & David Traver

  2. Program in Gene Function and Expression, University of Massachusetts Medical School, Worcester, 01605, Massachusetts, USA

    John C. Moore & Nathan D. Lawson

Authors
  1. Wilson K. Clements
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  2. Albert D. Kim
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Contributions

W.K.C. and D.T. designed all experiments. Whole-mount immunofluorescence, double fluorescence in situs, and Kaede-based fate mapping was performed by A.D.K. K.G.O. cloned and subcloned multiple constructs. J.C.M and N.L. generated Notch reporter lines. All other experiments were performed by W.K.C. The manuscript was written by W.K.C. and edited by N.L. and D.T., with critical input as described in the Acknowledgments.

Corresponding author

Correspondence to David Traver.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Information

The file contains Supplementary Figures 1-15 with legends and Supplementary Tables 1-9. (PDF 3823 kb)

Supplementary Movie 1

The movie shows timelapse imaging of GFP+ HSCs in the trunk region of untreated cd41:GFP transgenic animals captured at 1 frame per 3 minutes from 50 hpf to 75 hpf, documenting the behaviour of HSCs in this time period. (MOV 8601 kb)

Supplementary Movie 2

The movie shows timelapse imaging of lack of GFP+ HSCs in the trunk region of W16MO-injected cd41:GFP transgenic animals captured at 1 frame per 3 minutes from 50 hpf to 75 hpf, documenting decreased HSCs in this time period. (MOV 6967 kb)

Supplementary Movie 3

The movie shows timelapse imaging of GFP+ thymic immigrants in the head region of untreated cd41:GFP transgenic animals captured at 1 frame per 3 minutes from 50 hpf to 75 hpf, documenting the transition of HSCs to nascent T cells in this time period. (MOV 9791 kb)

Supplementary Movie 4

The movie shows timelapse imaging of lack of GFP+ thymic immigrants in the head region of W16MO-injected cd41:GFP transgenic animals captured at 1 frame per 3 minutes from 50 hpf to 75 hpf, documenting decreased transition of HSCs to nascent T cells in this time period. (MOV 6550 kb)

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Clements, W., Kim, A., Ong, K. et al. A somitic Wnt16/Notch pathway specifies haematopoietic stem cells. Nature 474, 220–224 (2011). https://doi.org/10.1038/nature10107

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