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The Lin28b–let-7–Hmga2 axis determines the higher self-renewal potential of fetal haematopoietic stem cells

Nature Cell Biology volume 15, pages 916–925 (2013)Cite this article

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Abstract

Mouse haematopoietic stem cells (HSCs) undergo a postnatal transition in several properties, including a marked reduction in their self-renewal activity. We now show that the developmentally timed change in this key function of HSCs is associated with their decreased expression of Lin28b and an accompanying increase in their let-7 microRNA levels. Lentivirus-mediated overexpression of Lin28 in adult HSCs elevates their self-renewal activity in transplanted irradiated hosts, as does overexpression of Hmga2, a well-established let-7 target that is upregulated in fetal HSCs. Conversely, HSCs from fetal Hmga2−/− mice do not exhibit the heightened self-renewal activity that is characteristic of wild-type fetal HSCs. Interestingly, overexpression of Hmga2 in adult HSCs does not mimic the ability of elevated Lin28 to activate a fetal lymphoid differentiation program. Thus, Lin28b may act as a master regulator of developmentally timed changes in HSC programs with Hmga2 serving as its specific downstream modulator of HSC self-renewal potential.

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Figure 1: Lin28b, let-7 targets and let-7 microRNAs are differentially expressed in fetal and adult HSCs.
Figure 2: Experimental design used to quantify the self-renewal activity of transplanted HSC populations.
Figure 3: Lin28 overexpression in ABM HSCs causes an increase in their expression of Hmga2.
Figure 4: Lin28 and Hmga2 overexpression increases the self-renewal activity of ABM HSCs.
Figure 5: Adult Hmga2 KO mice have reduced HSC numbers that match their reduced size.
Figure 6: Hmga2 is required for the high self-renewal activity of fetal HSCs.
Figure 7: Hmga2 overexpression in adult HSCs does not recapitulate the Lin28-mediated activation of fetal B-cell differentiation programs.
Figure 8: Developmental regulation of HSC self-renewal by the Lin28b–let-7–Hmga2 axis.

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Acknowledgements

This work was supported by grants from the National Cancer Institute of Canada (NCIC, with funds from the Terry Fox Run), the Canadian Institutes of Health Research (CIHR), the Canadian Cancer Society (grant 700374), and the Terry Fox Foundation. M.R.C. received a Vanier Canada Graduate Scholarship from CIHR, a Michael Smith Foundation for Health Research (MSFHR) Graduate Studentship and an MD/PhD Graduate Training Award from CIHR. S.B. received a University of British Columbia Graduate Studentship. C.B. received a fellowship from the Deutsche Forschungsgemeinschaft. D.J.H.F.K. received a CIHR Transplantation Research Training Award, a Frederick Banting and Charles Best Canada Graduate Scholarship, and a Vanier Canada Graduate Scholarship from CIHR. P.A.B. received a Kay Kendall Leukemia Fund Intermediate Fellowship from the UK. D.G.K. received studentships from CIHR and MSFHR. F.K. was supported by Deutsche Krebshilfe grant 109420, Fellowship 2010/04 from the European Hematology Association, and a Deutsche Forschungsgemeinschaft grant (KU2288/3-1). We thank support staff of the Flow Cytometry Facility at the Terry Fox Laboratory, the Animal Resource Centre and the Centre for Translational and Applied Genomics at the BC Cancer Agency, M. Hale and G. Edin for technical assistance, and M. Kardel, M. Makarem, A. Karsan and P. Hoodless for helpful comments.

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  1. Terry Fox Laboratory, BC Cancer Agency, Vancouver, British Columbia V5Z 1L3, Canada

    Michael R. Copley, Sonja Babovic, Claudia Benz, David J. H. F. Knapp, Philip A. Beer, David G. Kent, Stefan Wohrer, David Q. Treloar, Christopher Day, Keegan Rowe, Heidi Mader, Florian Kuchenbauer, R. Keith Humphries & Connie J. Eaves

  2. Departments of Medical Genetics and Medicine, University of British Columbia, British Columbia V6T 1Z3, Canada

    R. Keith Humphries & Connie J. Eaves

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  1. Michael R. Copley
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Contributions

M.R.C. performed all of the experiments with assistance from S.B. for qRT–PCR, intracellular flow cytometry and peritoneal cavity cell analyses, and F.K. for let-7 profiling. D.J.H.F.K. performed the analysis of Affymetrix array data. D.Q.T. collected ESLAM HSCs from E14.5 fetal livers for qRT–PCR analyses. H.M., K.R, C.D. and D.Q.T. performed the analyses of transplanted mice. M.R.C. and C.J.E. designed and interpreted all experiments with input from C.B., D.G.K., S.W. and R.K.H. M.R.C., C.B., P.A.B. and C.J.E. wrote the manuscript with final approval from all authors.

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Correspondence to Connie J. Eaves.

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Copley, M., Babovic, S., Benz, C. et al. The Lin28b–let-7–Hmga2 axis determines the higher self-renewal potential of fetal haematopoietic stem cells. Nat Cell Biol 15, 916–925 (2013). https://doi.org/10.1038/ncb2783

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