title: Glutaminase and poly(ADP-ribose) polymerase inhibitors suppress pyrimidine synthesis and VHL-deficient renal cancers.
abstract: 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.
authors.forename: Arimichi
authors.initials: A
authors.lastname: Okazaki
authors.forename: Paulo A
authors.initials: PA
authors.lastname: Gameiro
authors.forename: Danos
authors.initials: D
authors.lastname: Christodoulou
authors.forename: Laura
authors.initials: L
authors.lastname: Laviollette
authors.forename: Meike
authors.initials: M
authors.lastname: Schneider
authors.forename: Frances
authors.initials: F
authors.lastname: Chaves
authors.forename: Anat
authors.initials: A
authors.lastname: Stemmer-Rachamimov
authors.forename: Stephanie A
authors.initials: SA
authors.lastname: Yazinski
authors.forename: Richard
authors.initials: R
authors.lastname: Lee
authors.forename: Gregory
authors.initials: G
authors.lastname: Stephanopoulos
authors.forename: Lee
authors.initials: L
authors.lastname: Zou
authors.forename: Othon
authors.initials: O
authors.lastname: Iliopoulos
mesh_terms: D000818:Animals; D002292:Carcinoma, Renal Cell; D005971:Glutamates / Q000235:genetics / Q000378:metabolism; D005972:Glutaminase / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D005973:Glutamine / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007680:Kidney Neoplasms / Q000188:drug therapy* / Q000201:enzymology / Q000235:genetics / Q000473:pathology; D051379:Mice; D008819:Mice, Nude; D010793:Phthalazines / Q000494:pharmacology*; D010879:Piperazines / Q000494:pharmacology*; D000067856:Poly(ADP-ribose) Polymerase Inhibitors / Q000494:pharmacology*; D011065:Poly(ADP-ribose) Polymerases / Q000235:genetics / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D059807:S Phase Cell Cycle Checkpoints / Q000187:drug effects / Q000235:genetics; D051794:Von Hippel-Lindau Tumor Suppressor Protein / Q000235:genetics* / Q000378:metabolism; D023041:Xenograft Model Antitumor Assays
chemical_list: D005971:Glutamates; D010793:Phthalazines; D010879:Piperazines; D000067856:Poly(ADP-ribose) Polymerase Inhibitors; D017382:Reactive Oxygen Species; C088956:alpha-ketoglutamic acid; D005973:Glutamine; D051794:Von Hippel-Lindau Tumor Suppressor Protein; D011065:Poly(ADP-ribose) Polymerases; C579480:GLS protein, human; D005972:Glutaminase; C497446:VHL protein, human; C531550:olaparib