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Germline CBL mutations cause developmental abnormalities and predispose to juvenile myelomonocytic leukemia

Abstract

CBL encodes a member of the Cbl family of proteins, which functions as an E3 ubiquitin ligase. We describe a dominant developmental disorder resulting from germline missense CBL mutations, which is characterized by impaired growth, developmental delay, cryptorchidism and a predisposition to juvenile myelomonocytic leukemia (JMML). Some individuals experienced spontaneous regression of their JMML but developed vasculitis later in life. Importantly, JMML specimens from affected children show loss of the normal CBL allele through acquired isodisomy. Consistent with these genetic data, the common p.371Y>H altered Cbl protein induces cytokine-independent growth and constitutive phosphorylation of ERK, AKT and S6 only in hematopoietic cells in which normal Cbl expression is reduced by RNA interference. We conclude that germline CBL mutations have developmental, tumorigenic and functional consequences that resemble disorders that are caused by hyperactive Ras/Raf/MEK/ERK signaling and include neurofibromatosis type 1, Noonan syndrome, Costello syndrome, cardiofaciocutaneous syndrome and Legius syndrome.

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Figure 1: Autosomal dominant germline mutations in CBL are associated with a phenotype, GM-CSF hypersensitivity and vasculitis.
Figure 2: Consequences of splice site mutations in cDNA from individuals D347, D647 and I066.
Figure 3: p.371Y>H does not confer cytokine sensitivity or cytokine independent growth until silencing of mouse Cbl.
Figure 4: Cbl mutant proteins show prolonged protein turnover and are associated with increased phosphorylated EGFR upon EGF stimulation.

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Acknowledgements

We gratefully acknowledge the generous participation of the families included in this report. Supported in part by the US National Institutes of Health (CA113557 to M.L.L.); the V Foundation for Cancer Research (M.L.L. and B.S.B.); NIH/NCI (K08 CA103868 to B.S.B., R01 CA104282 to M.L.L. and B.S.B.); The Leukemia Lymphoma Society (6059-09, 2157-08 to M.L.L.); the Frank A. Campini Foundation (M.L.L. and B.S.B.); The Concern Foundation (B.S.B.); Deutsche Forschungsgemeinschaft (KR3473/1-1 to C.F.); Deutsche Krebshilfe (108220 to C.M.N. and C.F.); Deutsche José Carreras Leukämie-Stiftung (R08/19 to C.F.); the Canadian Cancer Society (16056 to M.O.); and the National Institute of General Medical Sciences (T32GM007618 to D.H.S.). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institute of General Medical Sciences or the National Institutes of Health.

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C.M.N. coordinated and collected clinical data from the subjects with EWOG-MDS and wrote the manuscript; M.W.K. collected clinical data, performed laboratory assays including sequencing, proliferation assays and prepared figures; D.H.S. performed laboratory assays including the shRNA experiments, the proliferation assays and protein blots; I.F. collected clinical data; M.E. collected subject samples and performed mutational analysis on highly purified populations of blood cells; N.J.B. contributed subject samples; S.B. performed ubiquitin assays; J.Z.F. and K.M.S. contributed subject samples; T.A.G. performed RNA isolation and cDNA sequencing; P.M. contributed subject samples; I.S., G.K., S.C., P.J.L., C.K. and P.G.S. contributed subject samples; A.H. provided age-matched control samples from children with asthma; M.S. performed mutational analysis; J.S., M.M.v.H., H.H. and F.L. contributed subject samples and collected clinical data; D.S. collected clinical data; S.A. performed colony assays and performed cDNA sequencing; L.C. collected clinical data; R.C.R. and S.S.S. performed ubiquitylation assays; M.O. supervised the ubiquitylation assays and wrote the manuscript; B.S.B. oversaw the shRNA and cell proliferative experiments and wrote the manuscript; C.F. collected clinical data and performed sequencing; M.L.L. coordinated and collected clinical and laboratory data from the USA, oversaw all of the laboratory work, coordinated the data and wrote the manuscript.

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Correspondence to Charlotte M Niemeyer or Mignon L Loh.

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Niemeyer, C., Kang, M., Shin, D. et al. Germline CBL mutations cause developmental abnormalities and predispose to juvenile myelomonocytic leukemia. Nat Genet 42, 794–800 (2010). https://doi.org/10.1038/ng.641

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