NRF2 activation induces NADH-reductive stress, providing a metabolic vulnerability in lung cancer

• Published in: Cell metabolism 2023-03, Vol.35 (3), p.487-503.e7
• Main Authors: Weiss-Sadan, Tommy, Ge, Maolin, Hayashi, Makiko, Gohar, Magdy, Yao, Cong-Hui, de Groot, Adriaan, Harry, Stefan, Carlin, Alexander, Fischer, Hannah, Shi, Lei, Wei, Ting-Yu, Adelmann, Charles H., Wolf, Konstantin, Vornbäumen, Tristan, Dürr, Benedikt R., Takahashi, Mariko, Richter, Marianne, Zhang, Junbing, Yang, Tzu-Yi, Vijay, Vindhya, Fisher, David E., Hata, Aaron N., Haigis, Marcia C., Mostoslavsky, Raul, Bardeesy, Nabeel, Papagiannakopoulos, Thales, Bar-Peled, Liron
• Format: Article
• Language: English
• Subjects: Carcinoma, Non-Small-Cell Lung - metabolism; Cell Line, Tumor; functional genomic; Humans; Kelch-Like ECH-Associated Protein 1 - metabolism; Lung Neoplasms - metabolism; NAD - metabolism; NADH/NAD; NF-E2-Related Factor 2 - genetics; NF-E2-Related Factor 2 - metabolism; non-small cell lung cancer; NRF2-KEAP1 pathway; oxidative phosphorylation; Oxidative Stress - genetics; reductive stress; Signal Transduction
• ISSN: 1550-4131; 1932-7420; 1932-7420
• DOI: 10.1016/j.cmet.2023.01.012

Multiple cancers regulate oxidative stress by activating the transcription factor NRF2 through mutation of its negative regulator, KEAP1. NRF2 has been studied extensively in KEAP1-mutant cancers; however, the role of this pathway in cancers with wild-type KEAP1 remains poorly understood. To answer this question, we induced NRF2 via pharmacological inactivation of KEAP1 in a panel of 50+ non-small cell lung cancer cell lines. Unexpectedly, marked decreases in viability were observed in >13% of the cell lines—an effect that was rescued by NRF2 ablation. Genome-wide and targeted CRISPR screens revealed that NRF2 induces NADH-reductive stress, through the upregulation of the NAD+-consuming enzyme ALDH3A1. Leveraging these findings, we show that cells treated with KEAP1 inhibitors or those with endogenous KEAP1 mutations are selectively vulnerable to Complex I inhibition, which impairs NADH oxidation capacity and potentiates reductive stress. Thus, we identify reductive stress as a metabolic vulnerability in NRF2-activated lung cancers. [Display omitted] •Small molecule and CRISPR screens identify KEAP1 as a dependency in lung cancer•The glycolytic vs. oxidative nature of cancer cells underlies KEAP1 dependency•NRF2 activation induces NADH-reductive stress through the upregulation of ALDH3A1•NADH-reductive stress is a metabolic liability in NRF2-activated cancers A subset of lung cancer cell lines is sensitive to activation of the transcription factor NRF2. The metabolic state of a cancer cell dictates sensitivity to NRF2, whose activation results in a NADH/NAD+ imbalance. As a result, the blockade of mitochondrial Complex I is a synthetic lethality in NRF2-activated lung cancers.