Identification of a set of genes potentially responsible for resistance to ferroptosis in lung adenocarcinoma cancer stem cells.
Francesca Ascenzi, Antonella Esposito, Sara Bruschini, Valentina Salvati, Claudia De Vitis, Valeria De Arcangelis, Giulia Ricci, Angiolina Catizione, Simona di Martino, Simonetta Buglioni, Massimiliano Bassi, Federico Venuta, Francesca De Nicola, Alice Massacci, Isabella Grassucci, Matteo Pallocca, Alberto Ricci, Maurizio Fanciulli, Gennaro Ciliberto, Rita Mancini
Author Information
Francesca Ascenzi: Translational Oncology Research Unit, IRCCS Regina Elena National Cancer Institute, Rome, Italy.
Antonella Esposito: Department of Experimental and Clinical Medicine, Magna Graecia University of Catanzaro, Catanzaro, Italy.
Sara Bruschini: Department of Clinical and Molecular Medicine, Sant' Andrea Hospital-Sapienza University of Rome, Rome, Italy.
Valentina Salvati: Preclinical Models and New Therapeutic Agents Unit, IRCCS Regina Elena National Cancer Institute, Rome, Italy.
Claudia De Vitis: Department of Clinical and Molecular Medicine, Sant' Andrea Hospital-Sapienza University of Rome, Rome, Italy.
Valeria De Arcangelis: Department of Clinical and Molecular Medicine, Sant' Andrea Hospital-Sapienza University of Rome, Rome, Italy.
Giulia Ricci: Department of Experimental Medicine, Università Degli Studi Della Campania Luigi Vanvitelli, Naples, Italy.
Angiolina Catizione: Department of Anatomy, Histology, Forensic-Medicine and Orthopedics, Sapienza University of Rome, Rome, Italy. ORCID
Simona di Martino: Pathology Unit, IRCCS-Regina Elena National Cancer Institute, Rome, Italy.
Simonetta Buglioni: Pathology Unit, IRCCS-Regina Elena National Cancer Institute, Rome, Italy.
Massimiliano Bassi: Thoracic Surgery Unit, Sapienza University of Rome, Rome, Italy. ORCID
Federico Venuta: Thoracic Surgery Unit, Sapienza University of Rome, Rome, Italy.
Francesca De Nicola: SAFU Laboratory, IRCCS Regina Elena National Cancer Institute, Rome, Italy.
Alice Massacci: Biostatistics, Bioinformatics and Clinical Trial Center, IRCCS Regina Elena National Cancer Institute, Rome, Italy.
Isabella Grassucci: Biostatistics, Bioinformatics and Clinical Trial Center, IRCCS Regina Elena National Cancer Institute, Rome, Italy.
Matteo Pallocca: Institute of Experimental Endocrinology and Oncology, National Research Council, Naples, Italy.
Alberto Ricci: Respiratory Unit, Sant'Andrea Hospital, Sapienza University of Rome, Rome, Italy.
Maurizio Fanciulli: SAFU Laboratory, IRCCS Regina Elena National Cancer Institute, Rome, Italy. ORCID
Gennaro Ciliberto: Scientific Direction, IRCCS Regina Elena National Cancer Institute, Rome, Italy. ORCID
Rita Mancini: Department of Clinical and Molecular Medicine, Sant' Andrea Hospital-Sapienza University of Rome, Rome, Italy. rita.mancini@uniroma1.it. ORCID
Scientific literature supports the evidence that cancer stem cells (CSCs) retain inside low reactive oxygen species (ROS) levels and are, therefore, less susceptible to cell death, including ferroptosis, a type of cell death dependent on iron-driven lipid peroxidation. A collection of lung adenocarcinoma (LUAD) primary cell lines derived from malignant pleural effusions (MPEs) of patients was used to obtain 3D spheroids enriched for stem-like properties. We observed that the ferroptosis inducer RSL3 triggered lipid peroxidation and cell death in LUAD cells when grown in 2D conditions; however, when grown in 3D conditions, all cell lines underwent a phenotypic switch, exhibiting substantial resistance to RSL3 and, therefore, protection against ferroptotic cell death. Interestingly, this phenomenon was reversed by disrupting 3D cells and growing them back in adherence, supporting the idea of CSCs plasticity, which holds that cancer cells have the dynamic ability to transition between a CSC state and a non-CSC state. Molecular analyses showed that ferroptosis resistance in 3D spheroids correlated with an increased expression of antioxidant genes and high levels of proteins involved in iron storage and export, indicating protection against oxidative stress and low availability of iron for the initiation of ferroptosis. Moreover, transcriptomic analyses highlighted a novel subset of genes commonly modulated in 3D spheroids and potentially capable of driving ferroptosis protection in LUAD-CSCs, thus allowing to better understand the mechanisms of CSC-mediated drug resistance in tumors.
