Abstract
Mutations in the granulocyte colony-stimulating factor receptor (G-CSF-R) gene leading to a truncated protein have been identified in a cohort of neutropenia patients highly predisposed to acute myeloid leukemia. Such mutations act in a dominant manner resulting in hyperproliferation but impaired differentiation in response to G-CSF. This is due, at least in part, to defective internalization and loss of binding sites for several negative regulators, leading to sustained receptor activation. However, those signaling pathways responsible for mediating the hyperproliferative function have remained unclear. In this study, analysis of an additional G-CSF-R mutant confirmed the importance of residues downstream of Box 2 as important contributors to the sustained proliferation. However, maximal proliferation correlated with the ability to robustly activate signal transducer and activator of transcription (STAT) 5 in a sustained manner, whereas co-expression of dominant-negative STAT5, but not dominant-negative STAT3, was able to inhibit G-CSF-stimulated proliferation from a truncated receptor. Furthermore, a Janus kinase (JAK) inhibitor also strongly reduced the proliferative response, whereas inhibitors of mitogen-activated protein kinase/extracellular signal-regulated kinase kinase (MEK) or phosphatidylinositol (PI) 3-kinase reduced proliferation to a lesser degree. These data suggest that sustained JAK2/STAT5 activation is a major contributor to the hyperproliferative function of truncated G-CSF receptors, with pathways involving MEK and PI 3-kinase playing a reduced role.
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Acknowledgements
We thank Cristal Peck for expert help with paper preparation, Dora McPhee, Louise Wangerek and Michelle Hookham for technical assistance, as well as Mirjiam Hermans, Jim Johnston, Karim Dib and Massimo Gadina for reagents. This work was supported by grants from the KWF Kankerbestrijding, the NWO and the Deakin University Central Research Grants Scheme. ACW was variously supported by an EMBO Long Term Fellowship, a Sylvia and Charles Viertel Senior Medical Research Fellowship and a Deakin University International Study Program Grant.
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Gits, J., van Leeuwen, D., Carroll, H. et al. Multiple pathways contribute to the hyperproliferative responses from truncated granulocyte colony-stimulating factor receptors. Leukemia 20, 2111–2118 (2006). https://doi.org/10.1038/sj.leu.2404448
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DOI: https://doi.org/10.1038/sj.leu.2404448