Redox reprogramming protects colon cancer cells from disulfidptosis under glucose deprivation

Cancer cells thrive in the tumor microenvironment by reprogramming their metabolism and adapting to severe nutritional stress. Although some cancer cells can withstand prolonged glucose deprivation, the molecular mechanisms underlying this resilience remain poorly understood. In this study, we investigated how glucose deprivation-resistant (GDR) colon cancer cells survive sustained glucose starvation, focusing on their ability to counteract oxidative stress and disulfidptosis, a recently described form of cell death driven by disulfide stress.We established long-term glucose-deprived cultures of colon cancer cells and compared quiescent populations with GDR cells that had resumed proliferation. Metabolic rewiring was assessed using Seahorse XF metabolic flux analysis and LC-MS-based metabolomics to map energy production pathways and metabolite changes. RNA sequencing was performed to identify transcriptional alterations, including those associated with disulfidptosis. Cytoskeletal integrity was evaluated by phalloidin staining, and cell viability was measured using the resazurin assay. To test whether redox support could restore survival, we conducted rescue experiments with antioxidants such as N-acetylcysteine and S-adenosylmethionine, as well as thiol-reducing agents including 2-mercaptoethanol and dithiothreitol, under both glucose-deprived and docetaxel-induced stress conditions.We found that upon glucose depletion, a subpopulation of colon cancer cells enters a dormant state characterized by elevated ROS, loss of mitochondrial membrane potential, extensive DNA damage, and cytoskeletal collapse. In contrast, the surviving GDR cells adapt by shifting from glycolysis to oxidative phosphorylation (OXPHOS). Metabolomic profiling revealed increased levels of TCA cycle intermediates and a pronounced enrichment of redox-active metabolites, including cystine, glutamate, glutamine, gamma-glutamylcysteine, reduced GSH, and oxidized GSSG. Transcriptomic analyses further supported this metabolic transition, showing marked upregulation of disulfidptosis-associated genes. Functionally, these adaptations enhanced redox buffering capacity and reduced sensitivity to disulfide-induced cytotoxicity. Notably, supplementation with antioxidants and thiol-reducing agents preserved cytoskeletal integrity and improved overall cell viability, indicating that strengthening redox homeostasis is sufficient to counteract disulfidptosis, even under the combined stresses of chemotherapy and glucose deprivation.In conclusion, GDR colon cancer cells evade disulfidptosis by reprogramming metabolism toward OXPHOS and enhancing redox defense mechanisms that maintain cytoskeletal integrity. Targeting these adaptive redox pathways may provide a promising strategy to eliminate cells that survive glucose deprivation through metabolic plasticity.