EglN3 hydroxylase stabilizes BIM-EL linking VHL type 2C mutations to pheochromocytoma pathogenesis and chemotherapy resistance

Significance Different mutations of the von Hippel–Lindau (VHL) tumor suppressor gene cause different subtypes of VHL hereditary cancer syndrome. Dysregulation of VHL’s canonical substrate, HIFα, cannot fully explain the complex genotype/phenotype manifestation within VHL disease. We describe an oxygen-sensitive function of VHL that regulates hydroxylated BIM-EL protein stability. Type 2C VHL mutants, which cause pheochromocytoma and paraganglioma (PCC/PGL) despite repressing HIFα, destabilize BIM-EL. Other genetic mutations linked to PCC/PGL, similar to loss of BIM-EL, protect from apoptosis upon nerve growth factor withdrawal. Dysregulation of BIM-EL might therefore contribute to the pathogenesis of VHL-related PCC/PGL. Loss of BIM-EL expression can lead to chemotherapy resistance in other VHL-related neoplasms and renders clear cell renal cell carcinoma cells insensitive to cisplatin. Despite the discovery of the oxygen-sensitive regulation of HIFα by the von Hippel–Lindau (VHL) protein, the mechanisms underlying the complex genotype/phenotype correlations in VHL disease remain unknown. Some germline VHL mutations cause familial pheochromocytoma and encode proteins that preserve their ability to down-regulate HIFα. While type 1, 2A, and 2B VHL mutants are defective in regulating HIFα, type 2C mutants encode proteins that preserve their ability to down-regulate HIFα. Here, we identified an oxygen-sensitive function of VHL that is abolished by VHL type 2C mutations. We found that BIM-EL, a proapoptotic BH3-only protein, is hydroxylated by EglN3 and subsequently bound by VHL. VHL mutants fail to bind hydroxylated BIM-EL, regardless of whether they have the ability to bind hydroxylated HIFα or not. VHL binding inhibits BIM-EL phosphorylation by extracellular signal-related kinase (ERK) on serine 69. This causes BIM-EL to escape from proteasomal degradation, allowing it to enhance EglN3-induced apoptosis. BIM-EL was rapidly degraded in cells lacking wild-type VHL or in which EglN3 was inactivated genetically or by lack of oxygen, leading to enhanced cell survival and chemotherapy resistance. Combination therapy using ERK inhibitors, however, resensitizes VHL- and EglN3-deficient cells that are otherwise cisplatin-resistant.

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