Abstract
Auranofin Synergistically Sensitizes KEAP1 Mutant and WT NSCLCs to P-AscH--induced Cell Killing by a Thiol and H2O2 Dependent Mechanism
Free radical biology & medicine, Vol.240(Suppl 1), p.S42
11/2025
DOI: 10.1016/j.freeradbiomed.2025.10.122
Abstract
Lung cancer is one of the most common and deadliest cancers in the United States, with 80-85% non-small cell lung cancers (NSCLC). 11-27% of the NSCLCs harbor KEAP1 (Kelch-like ECH-associated protein 1) mutations, resulting in activation of nuclear factor-erythroid factor 2 (NRF2) transcription factor, leading to upregulation of thioredoxin/glutathione metabolism, alterations in iron metabolism, and increased resistance to radio-chemotherapy, immunotherapy, and tyrosine kinase inhibitors. Treatment with Auranofin (thioredoxin reductase inhibitor) or pharmacological ascorbate (P-AscH-) causes cytotoxicity in cancer cells by a process dependent on thiol and H2O2 metabolism. The hypothesis that inhibiting thioredoxin reductase with Auranofin could sensitize KEAP1 mutant and WT NSCLCs to P-AscH- by a mechanism dependent on thiol metabolism and H2O2 was tested. KEAP1 mutant (A549, H2030, H460) and KEAP1 WT (CALU-6, H1299, H1299T, and H292) were treated with Auranofin (0-2 uM for 24 hr) or P-AscH- (0-40 pmol/cell for 1 hr) followed by clonogenic survival assay. Normal non-immortalized lung epithelial cells (HBEpC) were also treated with P-AscH- (0-40 pmol/cell for 1 hr) for comparison. All KEAP1 mutant NSCLCs were more resistant to clonogenic cell killing by both Auranofin and P-AscH- compared to KEAP1 WT NSCLCs. P-AscH- was not toxic to HBEpC (0-40 pmol/cell). Auranofin pretreatment for 24 hours (1 uM) synergistically sensitized KEAP1 mutant NSCLC cells to P-AscH- (10-40 pmol/cell, 1 hr) which was reversed in A549 by simultaneous treatment with the thiol antioxidant, N-Acetyl-L-cysteine (NAC, 24 hours). Auranofin pretreatment for 24 hours (0.5 uM) synergistically sensitized KEAP1 WT NSCLC cells to 1 hr exposure to P-AscH- (5-20 pmol/cell) which was reversed in H1299T by doxycycline inducible over expression of catalase. These findings support the hypothesis that KEAP1 mutant NSCLCs are more resistant to P-AscH- and targeting peroxide metabolism with Auranofin could enhance NSCLC therapeutic outcomes using P-AscH- (supported by P01 CA217797, T32 CA078586, P01 CA244091).
Details
- Title: Subtitle
- Auranofin Synergistically Sensitizes KEAP1 Mutant and WT NSCLCs to P-AscH--induced Cell Killing by a Thiol and H2O2 Dependent Mechanism
- Creators
- Sei ShoCollin D. HeerCasey F. PulliamAnn Tomanek-ChalkleyDaniel MarrenShafin WasimiAsya LengelMadi GentlesAdam J. DupuyDouglas R. Spitz
- Resource Type
- Abstract
- Publication Details
- Free radical biology & medicine, Vol.240(Suppl 1), p.S42
- DOI
- 10.1016/j.freeradbiomed.2025.10.122
- ISSN
- 0891-5849
- eISSN
- 1873-4596
- Publisher
- Elsevier
- Language
- English
- Date published
- 11/2025
- Academic Unit
- Anatomy and Cell Biology; Pathology; Radiation Oncology; Fraternal Order of Eagles Diabetes Research Center
- Record Identifier
- 9985024257202771
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