NRF2（核因子红系2有关因子2）是健康组织中保护性反应的主要调控因子。然而，当它在肿瘤细胞中活跃时，可能导致药物耐药性。内源性NRF2 抑制剂KEAP1结合NRF2并将其重定向至蛋白酶降解，因此KEAP1/NRF2相互作用对于维持NRF2在基础水平上至关重要。一些与临床相关的KEAP1突变显示破坏了这一关键的KEAP1/NRF2相互作用，导致NRF2水平升高和药物耐药性。在这里，我们描述了一种小分子NRF2抑制剂（R16），其选择性结合KEAP1突变体，并通过修复被破坏的KEAP1/NRF2相互作用恢复其抑制NRF2的功能。R16显著增加了KEAP1突变体肿瘤细胞对顺铂和吉非替尼的敏感性，但对于野生型KEAP1细胞则不具有这种效应，并在KEAP1 G333C突变的异种移植物中增加了对顺铂的敏感性。我们开发了一种基于BRET2的生物传感器系统来检测KEAP1/NRF2相互作用并对KEAP1突变进行分类。这一策略可通过临床肿瘤分子分析鉴定药物耐药的KEAP1体细胞突变。版权所有© 2023 作者。由Elsevier Inc.出版保留所有权利。

NRF2 (nuclear factor erythroid 2-related factor 2) is a master regulator of protective responses in healthy tissues. However, when it is active in tumor cells, it can result in drug resistance. KEAP1, the endogenous NRF2 inhibitor, binds NRF2 and redirects it to proteasomal degradation, so the KEAP1/NRF2 interaction is critical for maintaining NRF2 at a basal level. A number of clinically relevant KEAP1 mutations were shown to disrupt this critical KEAP1/NRF2 interaction, leading to elevated NRF2 levels and drug resistance. Here, we describe a small-molecule NRF2 inhibitor, R16, that selectively binds KEAP1 mutants and restores their NRF2-inhibitory function by repairing the disrupted KEAP1/NRF2 interactions. R16 substantially sensitizes KEAP1-mutated tumor cells to cisplatin and gefitinib, but does not do so for wild-type KEAP1 cells, and sensitizes KEAP1 G333C-mutated xenograft to cisplatin. We developed a BRET2-based biosensor system to detect the KEAP1/NRF2 interaction and classify KEAP1 mutations. This strategy would identify drug-resistant KEAP1 somatic mutations in clinical molecular profiling of tumors.Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.