Original Article | Published:

Transcriptional control and signal transduction, cell cycle

Mass cytometry analysis reveals hyperactive NF Kappa B signaling in myelofibrosis and secondary acute myeloid leukemia

Leukemia volume 31, pages 19621974 (2017) | Download Citation

Abstract

Myeloproliferative neoplasms (MPNs) feature a malignant clone containing the JAK2 V617F mutation, or another mutation causing dysregulated JAK2 kinase activity. The multiple disease phenotypes of MPNs, and their tendency to transform phenotypically, suggest pathophysiologic heterogeneities beyond a common phenomenon of JAK2 hyperactivation. JAK2 has the potential to activate multiple other signaling molecules, either directly through downstream effectors, or indirectly through induction of target gene expression. We have interrogated myeloproliferative signaling in myelofibrosis (MF) and secondary acute myeloid leukemia (sAML) patient samples using mass cytometry, which allows the quantitative measurement of multiple signaling molecules simultaneously at the single-cell level, in cell populations representing a nearly complete spectrum of hematopoiesis. MF and sAML malignant cells demonstrated a high prevalence of hyperactivation of the JAK-STAT, MAP kinase, PI3 kinase and NFκB signaling pathways. Constitutive NFκB signaling was evident across MF and sAML patients. A supporting gene set enrichment analysis (GSEA) of MF showed many NFκB target genes to be expressed above normal levels in MF patient CD34+ cells. NFκB inhibition suppressed colony formation from MF CD34+ cells. This study indicates that NFκB signaling contributes to human myeloproliferative disease and is abnormally activated in MF and sAML.

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Acknowledgements

This work was supported by NIH grants K08HL106576 (Oh), K12HL087107 (Oh), and T32HL007088 (Engle, Fisher). This research was also supported by an American Cancer Society Postdoctoral Fellowship (Fisher). This work was also supported by a Doris Duke-Damon Runyon Clinical Investigator Award (Oh), BRIGHT Institute Pilot Research Project Award (Oh), Burroughs Wellcome Fund Collaborative Research Travel Grant (Oh), Sidney Kimmel Scholar Award (Oh), Leukemia Research Foundation New Investigator Award (Oh), and American Cancer Society Institutional Research Grant (Oh). Additional support was provided by the Washington University Institute of Clinical and Translational Sciences grant UL1TR000448 from the National Center for Advancing Translational Sciences of NIH. Support for patient sample collection and processing was provided by NIH grant P01CA101937. These studies were supported in part by funding provided by Incyte Corporation. Technical support was provided by the Alvin J Siteman Cancer Center Tissue Procurement Core Facility, Flow Cytometry Core, and Immunomonitoring Laboratory, which are supported by NCI Cancer Center Support Grant P30CA91842. The Immunomonitoring Laboratory is also supported by the Andrew M and Jane M Bursky Center for Human Immunology and Immunotherapy Programs. We thank C Miner for assistance with mass cytometry experiments, D Moore and K Luber for assistance with clinical samples and data, and D Bender and C Wilson for assistance with cytokine assays.

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Affiliations

  1. Division of Hematology, Department of Medicine, Washington University School of Medicine, St. Louis, MO, USA

    • D A C Fisher
    • , E K Engle
    • , M C Fulbright
    • , T B Collins
    • , S Bandyopadhyay
    • , A Zhou
    •  & S T Oh
  2. Center for Human Immunology and Immunotherapy Programs, Washington University School of Medicine, St. Louis, MO, USA

    • O Malkova
    •  & C A Miner
  3. Division of Hematology, Department of Medicine, Stanford University School of Medicine, Stanford, CA, USA

    • G K Behbehani
  4. Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, CA, USA

    • G P Nolan

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The authors declare no conflict of interest.

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Correspondence to S T Oh.

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https://doi.org/10.1038/leu.2016.377

Supplementary Information accompanies this paper on the Leukemia website (http://www.nature.com/leu)

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