Glutaminase and poly(ADP-ribose) polymerase inhibitors suppress pyrimidine synthesis and VHL-deficient renal cancers

J Clin Invest. 2017 May 1;127(5):1631-1645. doi: 10.1172/JCI87800. Epub 2017 Mar 27.


Many cancer-associated mutations that deregulate cellular metabolic responses to hypoxia also reprogram carbon metabolism to promote utilization of glutamine. In renal cell carcinoma (RCC), cells deficient in the von Hippel-Lindau (VHL) tumor suppressor gene use glutamine to generate citrate and lipids through reductive carboxylation (RC) of α-ketoglutarate (αKG). Glutamine can also generate aspartate, the carbon source for pyrimidine biosynthesis, and glutathione for redox balance. Here we have shown that VHL-/- RCC cells rely on RC-derived aspartate to maintain de novo pyrimidine biosynthesis. Glutaminase 1 (GLS1) inhibitors depleted pyrimidines and increased ROS in VHL-/- cells but not in VHL+/+ cells, which utilized glucose oxidation for glutamate and aspartate production. GLS1 inhibitor-induced nucleoside depletion and ROS enhancement led to DNA replication stress and activation of an intra-S phase checkpoint, and suppressed the growth of VHL-/- RCC cells. These effects were rescued by administration of glutamate, αKG, or nucleobases with N-acetylcysteine. Further, we observed that the poly(ADP-ribose) polymerase (PARP) inhibitor olaparib synergizes with GLS1 inhibitors to suppress the growth of VHL-/- cells in vitro and in vivo. This work describes a mechanism that explains the sensitivity of RCC tumor growth to GLS1 inhibitors and supports the development of therapeutic strategies for targeting VHL-deficient RCC.

MeSH terms

  • Animals
  • Carcinoma, Renal Cell
  • Glutamates / genetics
  • Glutamates / metabolism
  • Glutaminase / antagonists & inhibitors*
  • Glutaminase / genetics
  • Glutaminase / metabolism
  • Glutamine / genetics
  • Glutamine / metabolism
  • Humans
  • Kidney Neoplasms / drug therapy*
  • Kidney Neoplasms / enzymology
  • Kidney Neoplasms / genetics
  • Kidney Neoplasms / pathology
  • Mice
  • Mice, Nude
  • Phthalazines / pharmacology*
  • Piperazines / pharmacology*
  • Poly(ADP-ribose) Polymerase Inhibitors / pharmacology*
  • Poly(ADP-ribose) Polymerases / genetics
  • Poly(ADP-ribose) Polymerases / metabolism
  • Reactive Oxygen Species / metabolism
  • S Phase Cell Cycle Checkpoints / drug effects
  • S Phase Cell Cycle Checkpoints / genetics
  • Von Hippel-Lindau Tumor Suppressor Protein / genetics*
  • Von Hippel-Lindau Tumor Suppressor Protein / metabolism
  • Xenograft Model Antitumor Assays


  • Glutamates
  • Phthalazines
  • Piperazines
  • Poly(ADP-ribose) Polymerase Inhibitors
  • Reactive Oxygen Species
  • alpha-ketoglutamic acid
  • Glutamine
  • Von Hippel-Lindau Tumor Suppressor Protein
  • Poly(ADP-ribose) Polymerases
  • GLS protein, human
  • Glutaminase
  • VHL protein, human
  • olaparib