Ascenção, Kelly Oravecz, Olivia Szabo, Csaba 2026 Cancer stem cells (CSCs) contribute to therapeutic resistance, metastatic progression, and tumor recurrence, yet the metabolic pathways that sustain their survival remain incompletely understood. Here, we identify 3-mercaptopyruvate sulfurtransferase (3-MST), a hydrogen sulfide–producing enzyme encoded by MPST, as a metabolic dependency of colorectal CSCs. 3-MST expression was increased in human colorectal tumors and cancer cell lines and strongly correlated with proliferative capacity. HCT116-derived CSCs exhibited elevated 3-MST expression, increased hydrogen sulfide and reactive sulfur species production, altered membrane rigidity, and a metabolically restrained phenotype characterized by low basal oxidative phosphorylation and glycolysis. Genetic depletion of 3-MST preferentially impaired CSC proliferation, spheroid formation, stem-like properties, and migration, with less pronounced effects in differentiated parental cells. Pharmacological inhibition of 3-MST reproduced these effects across CSCs derived from several colorectal cancer cell lines and induced nearcomplete suppression of mitochondrial respiration and glycolytic activity. 3-MST inhibition also increased membrane fluidity, promoted cell death, and reduced CSC-derived tumor growth in mice. Integrated transcriptomic, proteomic, metabolomic, and lipidomic analyses demonstrated coordinated disruption of mitochondrial carbon metabolism, respiratory-chain maintenance, lipid desaturation, and membrane phospholipid homeostasis. These changes were accompanied by accumulation of free fatty acids and diacylglycerols and activation of antioxidants, integrated stress-response, endoplasmic-reticulum-stress, apoptotic, and p53- associated pathways. Ferroptosis-related molecular signatures were also enriched. These findings identify 3- MST as a critical regulator of colorectal CSC bioenergetics and membrane homeostasis and reveal a therapeutically exploitable metabolic vulnerability in treatment-resistant colorectal cancer. application/pdf https://folia.unifr.ch/global/documents/336721 https://folia.unifr.ch/documents/336721/files/1-s2.0-s2213231726003757-main_0.pdf https://folia.unifr.ch/documents/336721/files/1-s2.0-s2213231726003757-mmc1.pdf https://folia.unifr.ch/documents/336721/files/1-s2.0-s2213231726003757-mmc2.pdf https://folia.unifr.ch/documents/336721/files/1-s2.0-s2213231726003757-mmc3.pdf https://folia.unifr.ch/documents/336721/files/1-s2.0-s2213231726003757-mmc4.pdf https://folia.unifr.ch/documents/336721/files/1-s2.0-s2213231726003757-mmc5.pdf https://folia.unifr.ch/documents/336721/files/1-s2.0-s2213231726003757-mmc6.xlsx https://folia.unifr.ch/documents/336721/files/1-s2.0-s2213231726003757-mmc7.pdf eng https://www.sciencedirect.com/science/article/pii/S2213231726003757?via%3Dihub info:eu-repo/semantics/altIdentifier/doi/10.1016/j.redox.2026.104376 info:eu-repo/semantics/altIdentifier/issn/2213-2317 info:eu-repo/semantics/openAccess CC BY Redox Biology. - Elsevier BV. - 2026, vol. 97, p. 104376 Hydrogen sulfide 3-Mercaptopyruvate sulfurtransferase Mitochondria Bioenergetics Gasotransmitters Colon cancer info:eu-repo/classification/udc/616 Targeting 3-mercaptopyruvate sulfurtransferase selectively eliminates colorectal cancer stem cells http://purl.org/coar/resource_type/c_6501