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Molecular targets for therapy

Characterization of the leukemogenic potential of distal cytoplasmic CSF3R truncation and missense mutations

Leukemia volume 31, pages 2752–2760 (2017)Cite this article

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Abstract

An increasing number of variants of unknown significance are being identified in leukemia patients with the application of deep sequencing and these include CSF3R cytoplasmic mutations. Previous studies have demonstrated oncogenic potential of certain CSF3R truncation mutations prior to internalization motifs. However, the oncogenic potential of truncating the more distal region of CSF3R cytoplasmic domain as well as cytoplasmic missense mutations remains uncharacterized. Here we identified that CSF3R distal cytoplasmic truncation mutations (Q793–Q823) also harbored leukemogenic potential. Mechanistically, these distal cytoplasmic truncation mutations demonstrated markedly decreased receptor degradation, probably owing to loss of the de-phosphorylation domain (residues N818–F836). Furthermore, all truncations prior to Q823 demonstrated increased expression of the higher molecular weight CSF3R band, which is shown to be essential for the receptor surface expression and the oncogenic potential. We further demonstrated that sufficient STAT5 activation is essential for oncogenic potential. In addition, CSF3R K704A demonstrated transforming capacity due to interruption of receptor ubiquitination and degradation. In summary, we have expanded the region of the CSF3R cytoplasmic domain in which truncation or missense mutations exhibit leukemogenic capacity, which will be useful for evaluating the relevance of CSF3R mutations in patients and helpful in defining targeted therapy strategies.

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Acknowledgements

We thank Christopher Eide and Samantha Savage for sharing the CML sequence data. We also acknowledge Larry David and John Klimek for mass spectrometry analysis; Aurelie Snyder and Stefanie Kaech Petrie for ApoTome microscope assistance and Brianna Garcia for her help in the FACS sorting.

Author contributions

HZ designed and performed the experiment, analyzed the data and prepared the manuscript; ARS performed the experiment, analyzed the data and proofread the manuscript. SL helped perform the mouse colony-forming unit assay and proofread the manuscript; AR prepared DNA and RNA sequence samples. YS participated in the FACS experiments. SM helped maintain the cell culture and performed western blot experiment for the signaling pathway analysis. DB, BW and SKM performed and analyzed exome and RNA sequencing data. JWT designed the experiment, interpreted the data, wrote and revised the manuscript.

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Authors and Affiliations

  1. Department of Cell, Developmental & Cancer Biology, Oregon Health & Science University, Knight Cancer Institute, Portland, OR, USA,

    H Zhang, A Reister Schultz, S Luty, A Rofelty, S Means & J W Tyner

  2. Department of Molecular and Medical Genetics, Oregon Health & Science University, Portland, OR, USA,

    Y Su

  3. Division of Bioinformatics and Computational Biology, Department of Medical Informatics and Clinical Epidemiology, Oregon Health & Science University, Knight Cancer Institute, Portland, OR, USA,

    D Bottomly, B Wilmot & S K McWeeney

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  1. H Zhang
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Correspondence to J W Tyner.

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Competing interests

JWT receives research support from Agios Pharmaceuticals, Array Biopharma, Aptose Biosciences, AstraZeneca, Constellation Pharmaceuticals, Genentech, Gilead, Incyte Corporation, Janssen Pharmaceutica, Seattle Genetics, Syros, Takeda Pharmaceutical Company and is a consultant for Leap Oncology. The remaining authors declare no conflict of interest.

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Zhang, H., Reister Schultz, A., Luty, S. et al. Characterization of the leukemogenic potential of distal cytoplasmic CSF3R truncation and missense mutations. Leukemia 31, 2752–2760 (2017). https://doi.org/10.1038/leu.2017.126

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