Abstract
Chuvash polycythemia is a rare congenital form of polycythemia caused by homozygous R200W and H191D mutations in the VHL (von Hippel-Lindau) gene, whose gene product is the principal negative regulator of hypoxia-inducible factor. However, the molecular mechanisms underlying some of the hallmark abnormalities of Chuvash polycythemia, such as hypersensitivity to erythropoietin, are unclear. Here we show that VHL directly binds suppressor of cytokine signaling 1 (SOCS1) to form a heterodimeric E3 ligase that targets phosphorylated JAK2 (pJAK2) for ubiquitin-mediated destruction. In contrast, Chuvash polycythemia–associated VHL mutants have altered affinity for SOCS1 and do not engage with and degrade pJAK2. Systemic administration of a highly selective JAK2 inhibitor, TG101209, reversed the disease phenotype in VhlR200W/R200W knock-in mice, an experimental model that recapitulates human Chuvash polycythemia. These results show that VHL is a SOCS1-cooperative negative regulator of JAK2 and provide biochemical and preclinical support for JAK2-targeted therapy in individuals with Chuvash polycythemia.
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Acknowledgements
This work was supported by funds from the Canadian Cancer Society (020110 to M.O. and 018054 to M.S.I.), US National Institutes of Health R01 CA142794 (W.Y.K.), U54 CA151652 (W.Y.K.) and US Department of Defense grants PC094631 and W81XWH-08-1-0064 (W.Y.K). Plasmid encoding HA-SOCS1 was provided by R. Rottapel (Ontario Cancer Institute). Lentiviral reagents pMDG1.vsvg and psPAX2 were provided by L. Penn (Ontario Cancer Institute). JAK2 inhibitor TG101209 was provided by TargeGen. We thank the University North Carolina Animal Studies Core Facility and Mouse Phase 1 Unit for their assistance in the animal study. W.Y.K. is a Damon Runyon Merck clinical investigator. M.O. is a Canada Research Chair in molecular oncology.
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R.C.R. designed and carried out biochemical and mouse experiments. R.I.S. designed and carried out biochemical experiments. B.Z. carried out the mouse experiments. P.H., S.B., S.S.S., S.N.G., O.R., V.W.K.C. and L.M.B. carried out and assisted with the biochemical experiments. S.A.H., T.D.R. and M.M.H. assisted with the mouse experiments. D.L.B., D.A.C. and M.C.S. provided the reagents associated with the mouse experiments. W.Y.K. designed the mouse experiments. M.S.I. conceptualized the model and designed the experiments. M.O. conceptualized the model, designed the experiments and wrote the manuscript.
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Russell, R., Sufan, R., Zhou, B. et al. Loss of JAK2 regulation via a heterodimeric VHL-SOCS1 E3 ubiquitin ligase underlies Chuvash polycythemia. Nat Med 17, 845–853 (2011). https://doi.org/10.1038/nm.2370
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DOI: https://doi.org/10.1038/nm.2370
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