Abstract
Mutations in the DYNAMIN2 (DNM2) gene are frequently detected in human acute T-cell lymphoblastic leukemia (T-ALL), although the mechanisms linking these mutations to disease pathogenesis remain unknown. Using an ENU-based forward genetic screen for mice with erythroid phenotypes, we identified a heterozygous mouse line carrying a mutation in the GTPase domain of Dnm2 (Dnm2V265G) that induced a microcytic anemia. In vitro assays using the V265G mutant demonstrated loss of GTPase activity and impaired endocytosis that was comparable to other DNM2 mutants identified in human T-ALL. To determine the effects of DNM2 mutations in T-ALL, we bred the Dnm2V265G mice with the Lmo2 transgenic mouse model of T-ALL. Heterozygous Dnm2 mutants lacking the Lmo2 transgene displayed normal T-cell development, and did not develop T-ALL. In contrast, compound heterozygotes displayed an accelerated onset of T-ALL compared with mice carrying the Lmo2 oncogene alone. The leukemias from these mice exhibited a more immature immunophenotype and an expansion in leukemic stem cell numbers. Mechanistically, the Dnm2 mutation impaired clathrin-mediated endocytosis of the interleukin (IL)-7 receptor resulting in increased receptor density on the surface of leukemic stem cells. These findings suggest that DNM2 mutations cooperate with T-cell oncogenes by enhancing IL-7 signalling.
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Acknowledgements
We thank Geza Paukovics, Jeanne LeMasurier and Phil Donaldson from the AMREP Flow Cytometry Facility for assistance with flow cytometry. We also thank Stephen Cody and Iśka Carmichael from the Monash Micro Imaging platform, as well as Shilpa Bereeka, Loretta Cerruti, Thomas Fulford and Nhu-Y Nguyen for technical assistance. This work was supported by a Project Grant (1052313, MPM and DJC) from the Australian National Health and Medical Research Council (NHMRC), a Research Fellowship (#700153) from The Terry Fox Foundation (CST) and a Senior Medical Research Fellowship from the Sylvia and Charles Viertel Foundation (DJC).
Author contributions
CST and DJC designed research, analyzed data and wrote the manuscript; CST, FCB, JS, MC, SEL, SES and SKC performed research and analyzed data; MLT provided reagents; RA and MPM provided biological samples; PJR and SMJ provided reagents and designed research.
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Tremblay, C., Brown, F., Collett, M. et al. Loss-of-function mutations of Dynamin 2 promote T-ALL by enhancing IL-7 signalling. Leukemia 30, 1993–2001 (2016). https://doi.org/10.1038/leu.2016.100
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DOI: https://doi.org/10.1038/leu.2016.100
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