Torrente, LauraPrieto-Farigua, NicolasFalzone, AimeeElkins, Cody M.Boothman, David A.Haura, Eric B.DeNicola, Gina M.2020-03-312020-03-312020-02Torrente, L., Prieto-Farigua, N., Falzone, A., Elkins, C. M., Boothman, D. A., Haura, E. B., & DeNicola, G. M. (2020). Inhibition of TXNRD or SOD1 overcomes NRF2-mediated resistance to β-lapachone. Redox biology, 30, 101440. https://doi.org/10.1016/j.redox.2020.101440https://hdl.handle.net/1805/22434Alterations in the NRF2/KEAP1 pathway result in the constitutive activation of NRF2, leading to the aberrant induction of antioxidant and detoxification enzymes, including NQO1. The NQO1 bioactivatable agent β-lapachone can target cells with high NQO1 expression but relies in the generation of reactive oxygen species (ROS), which are actively scavenged in cells with NRF2/KEAP1 mutations. However, whether NRF2/KEAP1 mutations influence the response to β-lapachone treatment remains unknown. To address this question, we assessed the cytotoxicity of β-lapachone in a panel of NSCLC cell lines bearing either wild-type or mutant KEAP1. We found that, despite overexpression of NQO1, KEAP1 mutant cells were resistant to β-lapachone due to enhanced detoxification of ROS, which prevented DNA damage and cell death. To evaluate whether specific inhibition of the NRF2-regulated antioxidant enzymes could abrogate resistance to β-lapachone, we systematically inhibited the four major antioxidant cellular systems using genetic and/or pharmacologic approaches. We demonstrated that inhibition of the thioredoxin-dependent system or copper-zinc superoxide dismutase (SOD1) could abrogate NRF2-mediated resistance to β-lapachone, while depletion of catalase or glutathione was ineffective. Interestingly, inhibition of SOD1 selectively sensitized KEAP1 mutant cells to β-lapachone exposure. Our results suggest that NRF2/KEAP1 mutational status might serve as a predictive biomarker for response to NQO1-bioactivatable quinones in patients. Further, our results suggest SOD1 inhibition may have potential utility in combination with other ROS inducers in patients with KEAP1/NRF2 mutations.en-USAttribution 4.0 InternationalKEAP1Kelch-like ECH-Associated protein 1NAD(P)H dehydrogenase [quinone] 1NQO1NRF2NSCLCNon-small cell lung cancerNuclear factor erythroid 2-related factor 2ROSReactive oxygen speciesβ-LapachoneArticle