多种癌症通过突变其负调节因子KEAP1来激活转录因子NRF2，从而调节氧化应激。先前已经在KEAP1突变癌症中广泛研究了NRF2，但是在KEAP1野生型癌症中这一通路的作用仍然不清楚。为了回答这个问题，我们通过药理学的方法在50多个非小细胞肺癌细胞系中诱导NRF2，使KEAP1失活。意外地观察到超过13%的细胞系中细胞存活率显著降低——这种效应是通过NRF2消融来挽救的。全基因组和有针对性的CRISPR筛查揭示了NRF2通过上调NAD+消耗酶ALDH3A1诱导NADH还原应激。利用这些发现，我们发现KEAP1抑制剂处理的细胞或者具有内源性KEAP1突变的细胞在复合物I抑制剂的选择性作用下，NADH氧化能力受损，还原应激增强。因此我们确定NRF2活化的肺癌细胞存在还原应激代谢上的敏感性。版权所有©2023 Elsevier Inc.

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.Copyright © 2023 Elsevier Inc. All rights reserved.