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Gain-of-function of mutated C-CBL tumour suppressor in myeloid neoplasms

Abstract

Acquired uniparental disomy (aUPD) is a common feature of cancer genomes, leading to loss of heterozygosity. aUPD is associated not only with loss-of-function mutations of tumour suppressor genes1, but also with gain-of-function mutations of proto-oncogenes2. Here we show unique gain-of-function mutations of the C-CBL (also known as CBL) tumour suppressor that are tightly associated with aUPD of the 11q arm in myeloid neoplasms showing myeloproliferative features. The C-CBL proto-oncogene, a cellular homologue of v-Cbl, encodes an E3 ubiquitin ligase and negatively regulates signal transduction of tyrosine kinases3,4,5,6. Homozygous C-CBL mutations were found in most 11q-aUPD-positive myeloid malignancies. Although the C-CBL mutations were oncogenic in NIH3T3 cells, c-Cbl was shown to functionally and genetically act as a tumour suppressor. C-CBL mutants did not have E3 ubiquitin ligase activity, but inhibited that of wild-type C-CBL and CBL-B (also known as CBLB), leading to prolonged activation of tyrosine kinases after cytokine stimulation. c-Cbl-/- haematopoietic stem/progenitor cells (HSPCs) showed enhanced sensitivity to a variety of cytokines compared to c-Cbl+/+ HSPCs, and transduction of C-CBL mutants into c-Cbl-/- HSPCs further augmented their sensitivities to a broader spectrum of cytokines, including stem-cell factor (SCF, also known as KITLG), thrombopoietin (TPO, also known as THPO), IL3 and FLT3 ligand (FLT3LG), indicating the presence of a gain-of-function that could not be attributed to a simple loss-of-function. The gain-of-function effects of C-CBL mutants on cytokine sensitivity of HSPCs largely disappeared in a c-Cbl+/+ background or by co-transduction of wild-type C-CBL, which suggests the pathogenic importance of loss of wild-type C-CBL alleles found in most cases of C-CBL-mutated myeloid neoplasms. Our findings provide a new insight into a role of gain-of-function mutations of a tumour suppressor associated with aUPD in the pathogenesis of some myeloid cancer subsets.

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Figure 1: Common UPD on the 11q arm and C-CBL mutations in myeloid neoplasms.
Figure 2: Tumour-suppressor functions of wild-type C-CBL.
Figure 3: Inhibitory actions of C-CBL mutants on wild-type C-CBL.
Figure 4: Gain-of-function of mutant C-CBL augmented by loss of wild-type C-CBL.

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Primary accessions

Gene Expression Omnibus

Data deposits

Full copy number data for the 222 samples are accessible from the Gene Expression Omnibus public database (http://ncbi.nlm.nih.gov/geo/) with the accession number GSE15187.

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Acknowledgements

This work was supported by the Core Research for Evolutional Science and Technology, Japan Science and Technology Agency, a Grant-in-Aid from the Ministry of Health, Labor and Welfare of Japan and from the Ministry of Education, Culture, Sports, Science and Technology, and a grant from National Health Research Institute, Taiwan, NHRI-EX96-9434SI, and NIH-2R01CA026038-30. We thank W. Y. Langdon for providing a human C-CBL cDNA. A mast-cell cell line expressing c-KIT V3MC was a gift from M. F. Gurish. We also thank Y. Ogino and K. Fujita for their technical assistance.

Author Contributions M.S. and M.Kato performed microarray experiments and subsequent data analyses. T.S., T.Y., H.Honda and H.Hirai generated and analysed c-Cbl-null mice. M.S., M.Otsu, S.Y., M.N., K.K., N.G., M.Onodera, M.S.-Y. and H.N. conducted functional assays of C-CBL mutants. L.-Y.S., M.S., M.Kato, K.N., J.T. and A.T. performed mutation analysis. H.O. performed pathological analysis of c-Cbl-null mice. L.-Y.S., N.K., H.Harada, M.Kurokawa, S.C., H.M., H.P.K. and M.Omine prepared MDS specimens. M.S., M.Otsu, Y.H., K.O., H.M., H.N., L.-Y.S., H.P.K. and S.O. designed the overall study, and S.O. wrote the manuscript. All authors discussed the results and commented on the manuscript.

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Correspondence to Lee-Yung Shih or Seishi Ogawa.

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Sanada, M., Suzuki, T., Shih, LY. et al. Gain-of-function of mutated C-CBL tumour suppressor in myeloid neoplasms. Nature 460, 904–908 (2009). https://doi.org/10.1038/nature08240

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