The t(8;21) rearrangement, which creates the AML1-ETO fusion protein, represents the most common chromosomal translocation in acute myeloid leukemia (AML). Clinical data suggest that CBL mutations are a frequent event in t(8;21) AML, but the role of CBL in AML1-ETO-induced leukemia has not been investigated. In this study, we demonstrate that CBL mutations collaborate with AML1-ETO to expand human CD34+ cells both in vitro and in a xenograft model. CBL depletion by shRNA also promotes the growth of AML1-ETO cells, demonstrating the inhibitory function of endogenous CBL in t(8;21) AML. Mechanistically, loss of CBL function confers hyper-responsiveness to thrombopoietin and enhances STAT5/AKT/ERK/Src signaling in AML1-ETO cells. Interestingly, we found the protein tyrosine phosphatase UBASH3B/Sts-1, which is known to inhibit CBL function, is upregulated by AML1-ETO through transcriptional and miR-9-mediated regulation. UBASH3B/Sts-1 depletion induces an aberrant pattern of CBL phosphorylation and impairs proliferation in AML1-ETO cells. The growth inhibition caused by UBASH3B/Sts-1 depletion can be rescued by ectopic expression of CBL mutants, suggesting that UBASH3B/Sts-1 supports the growth of AML1-ETO cells partly through modulation of CBL function. Our study reveals a role of CBL in restricting myeloid proliferation of human AML1-ETO-induced leukemia, and identifies UBASH3B/Sts-1 as a potential target for pharmaceutical intervention.
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We thank Dr Seishi Ogawa, Dr Masashi Sanada, Dr Masafumi Onodera and Dr Karsten Spiekermann for plasmids. We thank the Flow Cytometry Core and the Mouse Core at Cincinnati Children's Hospital Medical Center for their help. This work was supported by a grant from the CancerFree Kids Foundation for Cancer Research (JCM and SG), an Institutional Clinical and Translational Science Award, NIH/NCRR Grant Number 1UL1RR026314-01, Translational Trials Development and Support Laboratory award (U.S.P.H.S. Grant Number MO1 RR 08084), a Center of Excellence in Molecular Hematology P30 award (DK090971), JSPS Postdoctoral Fellowship for Research Abroad (SG), CA178454 (JC), the Coleman Leukemia Research Foundation (CDB), CA180861 (CDB), CA101140 (CDB), CA140158 (CDB), grants from Leukaemia Lymphoma Research (12007, CB) and (12055, OH). AG and NNN were supported by a grant from the Leukemia and Lymphoma Society. JCM is a Leukemia and Lymphoma Society Scholar.
SG conceived the project, designed and performed the research, analyzed the data and wrote the paper. JS performed the research and analyzed the data. MS, SL and KAL assisted with experiments. AG and NNN actively participated in designing and assisting with the experiments regarding UBASH3B/Sts-1. JC assisted with the experiments regarding miR-9. SPW, CDB and DN analyzed expression profiles of CBF-AML patients. SA, AP, OH and CB performed ChIP-Seq and RNA-Seq analyses. TK analyzed the data. JCM secured funding, analyzed the data and participated in writing the paper.
The authors declare no conflict of interest.
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Goyama, S., Schibler, J., Gasilina, A. et al. UBASH3B/Sts-1-CBL axis regulates myeloid proliferation in human preleukemia induced by AML1-ETO. Leukemia 30, 728–739 (2016). https://doi.org/10.1038/leu.2015.275
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