Article Interferon-Alpha Decreases Cancer Stem Cell Properties and Modulates Exosomes in Malignant Melanoma García Ortega, María Belén Aparicio, Ernesto Griñán Lisón, Carmen Jiménez González, Gema López Ruiz, Elena Palacios Ferrer, José Luis Ruiz Alcalá, Gloria Boulaiz Tassi, Houria Perán, Macarena Hackenberg, Michael Marchal Corrales, Juan Antonio García Chaves, María Ángel Garcí Interferon-α Malignant melanoma Cancer stem cells Exosomes Biomarkers Malignant melanoma (MM) can spread to other organs and is resistant in part due to the presence of cancer stem cell subpopulations (CSCs). While a controversial high dose of interferon-alpha (IFN-α) has been used to treat non-metastatic high-risk melanoma, it comes with undesirable side effects. In this study, we evaluated the effect of low and high doses of IFN-α on CSCs by analyzing ALDH activity, side population and specific surface markers in established and patient-derived primary cell lines. We also assessed the clonogenicity, migration and tumor initiation capacities of IFN-α treated CSCs. Additionally, we investigated genomic modulations related to stemness properties using microRNA sequencing and microarrays. The effect of IFN-α on CSCs-derived exosomes was also analyzed using NanoSight and liquid chromatography (LC-HRMS)-based metabolomic analysis, among others. Our results showed that even low doses of IFN-α reduced CSC formation and stemness properties, and led to a significant decrease in the ability to form tumors in mice xenotransplants. IFN-α also modulated the expression of genes and microRNAs involved in several cancer processes and metabolomics of released exosomes. Our work suggests the utility of low doses of interferon, combined with the analysis of metabolic biomarkers, as a potential clinical approach against the aggressiveness of CSCs in melanoma. 2023-09-22T09:37:57Z 2023-09-22T09:37:57Z 2023-07-18 journal article García-Ortega, M.B.; Aparicio, E.; Griñán-Lisón, C.; Jiménez, G.; López-Ruiz, E.; Palacios, J.L.; Ruiz-Alcalá, G.; Alba, C.; Martínez, A.; Boulaiz, H.; et al. Interferon-Alpha Decreases Cancer Stem Cell Properties and Modulates Exosomes in Malignant Melanoma. Cancers 2023, 15, 3666. [https:// doi.org/10.3390/cancers15143666] https://hdl.handle.net/10481/84587 10.3390/cancers15143666 eng http://creativecommons.org/licenses/by/4.0/ open access Atribución 4.0 Internacional MDPI