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Acute Leukemias

Identification of a role for the nuclear receptor EAR-2 in the maintenance of clonogenic status within the leukemia cell hierarchy

Leukemia volume 25, pages 1687–1696 (2011)Cite this article

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Abstract

Identification of genes that regulate clonogenicity of acute myelogenous leukemia (AML) cells is hindered by the difficulty of isolating pure populations of cells with defined proliferative abilities. By analyzing the growth of clonal siblings in low passage cultures of the cell line OCI/AML4 we resolved this heterogeneous population into strata of distinct clonogenic potential, permitting analysis of the transcriptional signature of single cells with defined proliferative abilities. By microarray analysis we showed that the expression of the orphan nuclear receptor EAR-2 (NR2F6) is greater in leukemia cells with extensive proliferative capacity than in those that have lost proliferative ability. EAR-2 is expressed highly in long-term hematopoietic stem cells, relative to short-term hematopoietic stem and progenitor cells, and is downregulated in AML cells after induction of differentiation. Exogenous expression of EAR-2 increased the growth of U937 cells and prevented the proliferative arrest associated with terminal differentiation, and blocked differentiation of U937 and 32Dcl3 cells. Conversely, silencing of EAR-2 by short-hairpin RNA initiated terminal differentiation of these cell lines. These data identify EAR-2 as an important factor in the regulation of clonogenicity and differentiation, and establish that analysis of clonal siblings allows the elucidation of differences in gene expression within the AML hierarchy.

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Acknowledgements

We thank Deborah Hyam and Mary Barbara for excellent technical assistance, Dr Suzanne Kamel-Reid for providing patient specimens, and Zeynep Alkan and Dr Brian Leber for their critical reading of the manuscript. This work was funded by a generous donation from the estate of J Douglas Crashley, a Canadian Institutes of Health Research operating grant (MOP 42420), a HSC Foundation New Investigator Award to RAW, CIHR Clinician Scientist awards to HLA and RAW, operating grants from the CCSRI and Terry Fox Foundation and support from the Princess Margaret Hospital Foundation and the Ontario Ministry of Health and Long Term Care to NNI and a CIHR-Canada Graduate Scholarship, an Adel S Sedra Award of Excellence, a Dr Joe Connolly Memorial OSOTF Award, a Government of Ontario/Dr Dina Gordon Malkin Graduate Scholarship in Science and Technology and a Frank Fletcher Memorial OSOTF Award to CVI.

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Author notes

  1. N N Iscove and R A Wells: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada

    C V Ichim, N N Iscove & R A Wells

  2. Discipline of Molecular and Cellular Biology, Sunnybrook Research Institute, Toronto, Ontario, Canada

    C V Ichim & R A Wells

  3. Center for Innovative Cancer Research, Ottawa Hospital Research Institute, Ottawa, Ontario, Canada

    H L Atkins

  4. Department of Molecular and Cell Biology, Ontario Cancer Institute, Toronto, Ontario, Canada

    N N Iscove

  5. McEwen Center for Regenerative Medicine, Toronto, Ontario, Canada

    N N Iscove

  6. Campbell Family Institute for Cancer Research, Toronto, Ontario, Canada

    N N Iscove

  7. Department of Medicine, University of Toronto, Toronto, Ontario, Canada

    R A Wells

  8. Department of Medical Oncology, Myelodysplastic Syndromes Program, Odette Cancer Centre, Sunnybrook Health Sciences Centre, Toronto, Ontario, Canada

    R A Wells

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  1. C V Ichim
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Correspondence to R A Wells.

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Ichim, C., Atkins, H., Iscove, N. et al. Identification of a role for the nuclear receptor EAR-2 in the maintenance of clonogenic status within the leukemia cell hierarchy. Leukemia 25, 1687–1696 (2011). https://doi.org/10.1038/leu.2011.137

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