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Stem Cells

CITED2-mediated human hematopoietic stem cell maintenance is critical for acute myeloid leukemia

Leukemia volume 29, pages 625–635 (2015)Cite this article

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Abstract

As the transcriptional coactivator CITED2 (CBP/p300-interacting-transactivator-with-an ED-rich-tail 2) can be overexpressed in acute myeloid leukemia (AML) cells, we analyzed the consequences of high CITED2 expression in normal and AML cells. CITED2 overexpression in normal CD34+ cells resulted in enhanced hematopoietic stem and progenitor cell (HSPC) output in vitro, as well as in better hematopoietic stem cell (HSC) engraftability in NSG (NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ) mice. This was because of an enhanced quiescence and maintenance of CD34+CD38− HSCs, due in part to an increased expression of the cyclin-dependent kinase inhibitor CDKN1A. We demonstrated that PU.1 is a critical regulator of CITED2, as PU.1 repressed CITED2 expression in a DNA methyltransferase 3A/B (DNMT3A/B)-dependent manner in normal CD34+ cells. CD34+ cells from a subset of AML patients displayed higher expression levels of CITED2 as compared with normal CD34+ HSPCs, and knockdown of CITED2 in AML CD34+ cells led to a loss of long-term expansion, both in vitro and in vivo. The higher CITED2 expression resulted from reduced PU.1 activity and/or dysfunction of mutated DNMT3A/B. Collectively, our data demonstrate that increased CITED2 expression results in better HSC maintenance. In concert with low PU.1 levels, this could result in a perturbed myeloid differentiation program that contributes to leukemia maintenance.

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Acknowledgements

We acknowledge RJ van der Lei, H Moes and G Mesander for help with flow cytometry, and Amgen Inc. and Kirin for providing cytokines. We greatly appreciate the help of Dr A van Loon, Dr JJ Erwich and colleagues (Obstetrics departments from the Martini Hospital and UMCG) for collecting CB, Dr AB Mulder (Department of Laboratory Medicine) and Professor E van den Berg (Department of Genetics) for mutation analyzes and A Brouwers-Vos (Department of Experimental Hematology) for help with Illumina Bead Arrays. We also thank Dr T Plosch and M Zwiers from the Department of Obstetrics and Gynecology for their help with the pyrosequencing. This work was funded by an NWO VENI grant (91611105) awarded to HS.

Author Contributions

PMK, GB, BB, MR, JJ and HS performed experiments and analyzed data. GdH, JJS, EV and HS analyzed and discussed data. JJS, EV and HS designed the experiments and wrote the manuscript.

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Authors and Affiliations

  1. Department of Experimental Hematology, Cancer Research Center Groningen, University Medical Center Groningen, University of Groningen, Groningen, The Netherlands

    P M Korthuis, G Berger, B Bakker, M Rozenveld-Geugien, J Jaques, J J Schuringa, E Vellenga & H Schepers

  2. Department of Stem Cell Biology, European Research Institute for the Biology of Aging (ERIBA), University Medical Center Groningen, University of Groningen, Groningen, The Netherlands

    G de Haan

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  1. P M Korthuis
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Correspondence to H Schepers.

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Korthuis, P., Berger, G., Bakker, B. et al. CITED2-mediated human hematopoietic stem cell maintenance is critical for acute myeloid leukemia. Leukemia 29, 625–635 (2015). https://doi.org/10.1038/leu.2014.259

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