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

Proc Natl Acad Sci U S A. 2019 Aug 20;116(34):16997-17006. doi: 10.1073/pnas.1900748116. Epub 2019 Aug 2.

Abstract

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.

Keywords: BIM-EL; VHL; hydroxylation; pheochromocytoma; tumor suppression.

MeSH terms

  • Adrenal Gland Neoplasms* / drug therapy
  • Adrenal Gland Neoplasms* / genetics
  • Adrenal Gland Neoplasms* / metabolism
  • Adrenal Gland Neoplasms* / pathology
  • Animals
  • Apoptosis / drug effects
  • Apoptosis / genetics
  • Bcl-2-Like Protein 11 / genetics
  • Bcl-2-Like Protein 11 / metabolism*
  • Cell Hypoxia / drug effects
  • Cell Hypoxia / genetics
  • Cisplatin / pharmacology
  • Drug Resistance, Neoplasm / drug effects
  • Drug Resistance, Neoplasm / genetics*
  • Humans
  • Hydroxylation / drug effects
  • Hydroxylation / genetics
  • Hypoxia-Inducible Factor-Proline Dioxygenases / genetics
  • Hypoxia-Inducible Factor-Proline Dioxygenases / metabolism*
  • MAP Kinase Signaling System / drug effects
  • MAP Kinase Signaling System / genetics
  • Mice
  • Mice, Knockout
  • Mutation*
  • PC12 Cells
  • Pheochromocytoma* / drug therapy
  • Pheochromocytoma* / metabolism
  • Pheochromocytoma* / pathology
  • Proteolysis / drug effects
  • Rats
  • Von Hippel-Lindau Tumor Suppressor Protein / genetics
  • Von Hippel-Lindau Tumor Suppressor Protein / metabolism*

Substances

  • BCL2L11 protein, human
  • Bcl-2-Like Protein 11
  • Bcl2l11 protein, mouse
  • EGLN3 protein, human
  • Egln3 protein, rat
  • Hypoxia-Inducible Factor-Proline Dioxygenases
  • Von Hippel-Lindau Tumor Suppressor Protein
  • VHL protein, human
  • VHL protein, mouse
  • Cisplatin