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A role for Wnt signalling in self-renewal of haematopoietic stem cells

Abstract

Haematopoietic stem cells (HSCs) have the ability to renew themselves and to give rise to all lineages of the blood; however, the signals that regulate HSC self-renewal remain unclear. Here we show that the Wnt signalling pathway has an important role in this process. Overexpression of activated β-catenin expands the pool of HSCs in long-term cultures by both phenotype and function. Furthermore, HSCs in their normal microenvironment activate a LEF-1/TCF reporter, which indicates that HCSs respond to Wnt signalling in vivo. To demonstrate the physiological significance of this pathway for HSC proliferation we show that the ectopic expression of axin or a frizzled ligand-binding domain, inhibitors of the Wnt signalling pathway, leads to inhibition of HSC growth in vitro and reduced reconstitution in vivo. Furthermore, activation of Wnt signalling in HSCs induces increased expression of HoxB4 and Notch1, genes previously implicated in self-renewal of HSCs. We conclude that the Wnt signalling pathway is critical for normal HSC homeostasis in vitro and in vivo, and provide insight into a potential molecular hierarchy of regulation of HSC development.

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Figure 1: Activated β-catenin promotes growth of HSCs in vitro and maintains the immature phenotype of HSCs in long-term cultures.
Figure 2: Activated β-catenin-transduced HSCs reconstitute lethally irradiated mice and give rise to multiple haematopoietic lineages.
Figure 3: HSCs respond to Wnt signalling in native bone marrow microenvironment.
Figure 4: Inhibition of Wnt signalling reduces growth of HSCs in vitro and inhibits reconstitution in vivo.
Figure 5: HSCs expressing β-catenin upregulate HoxB4 and Notch1.

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Acknowledgements

We thank W. J. Nelson and A. Barth for providing the β-catenin construct; G. Nolan and L. Naldini for providing the retroviral and lentiviral packaging systems; F. Costantini for providing the axin construct; J. Nathans for the Fzd8-CRD IgG construct; H. Clevers and B. Vogelstein for providing the modified LEF-1/TCF elements used in the reporter constructs; A. Carlton for help with proliferation assays; and P. Feliciano for generation of retroviral constructs. We are grateful to L. Jerabek and P. Shahi for laboratory management and technical help; L. Hidalgo for animal care; S. Smith and V. Braunstein for preparation of antibodies; M. Cook for cell sorting; and M. Krangel for use of the Light Cycler. D.C.S. is a fellow of the American Cancer Society, L.A. is a fellow of the Canadian Institute of Health Research and R.N. is an investigator of the Howard Hughes Medical Institute. This work was supported by NIH grants awarded to I.L.W., and funds from the Cancer Research Institute, Leukemia Research Foundation and the NIH awarded to T.R.

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Correspondence to Tannishtha Reya.

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I.L.W. is a co-founder of a company, Cellerant Therapeutics, Inc., established 31 January 2003, that has interests in haematopoietic stem cell biology and expansion. It does not fund any of the research of I.L.W. at Stanford.

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Reya, T., Duncan, A., Ailles, L. et al. A role for Wnt signalling in self-renewal of haematopoietic stem cells. Nature 423, 409–414 (2003). https://doi.org/10.1038/nature01593

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