Altered fatty acid metabolism rewires cholangiocarcinoma stemness features
Giulia Lori,
Mirella Pastore,
Nadia Navari,
Benedetta Piombanti,
Richell Booijink,
Elisabetta Rovida,
Ignazia Tusa,
Monika Lewinska,
Jesper B. Andersen,
Tiziano Lottini,
Annarosa Arcangeli,
Maria Letizia Taddei,
Erica Pranzini,
Caterina Mancini,
Cecilia Anceschi,
Stefania Madiai,
Elena Sacco,
Stefano Rota,
Adriana Trapani,
Gennaro Agrimi,
Matteo Ramazzotti,
Paola Ostano,
Caterina Peraldo Neia,
Matteo Parri,
Fabrizia Carli,
Silvia Sabatini,
Amalia Gastaldelli,
Fabio Marra,
Chiara Raggi
Affiliations
Giulia Lori
Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy
Mirella Pastore
Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy
Nadia Navari
Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy
Benedetta Piombanti
Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy
Richell Booijink
Department of Biomaterial Science and Technology, University of Twente Enschede, The Netherlands
Elisabetta Rovida
Department of Experimental and Clinical Biomedical Sciences, University of Florence, Florence, Italy
Ignazia Tusa
Department of Experimental and Clinical Biomedical Sciences, University of Florence, Florence, Italy
Monika Lewinska
Biotech Research and Innovation Centre, Department of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark
Jesper B. Andersen
Biotech Research and Innovation Centre, Department of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark
Tiziano Lottini
Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy
Annarosa Arcangeli
Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy
Maria Letizia Taddei
Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy
Erica Pranzini
Department of Experimental and Clinical Biomedical Sciences, University of Florence, Florence, Italy
Caterina Mancini
Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy
Cecilia Anceschi
Department of Experimental and Clinical Biomedical Sciences, University of Florence, Florence, Italy
Stefania Madiai
Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy
Elena Sacco
Department of Biotechnology and Biosciences, University of Milano-Bicocca, Milan, Italy
Stefano Rota
Department of Biotechnology and Biosciences, University of Milano-Bicocca, Milan, Italy
Adriana Trapani
Department of Pharmacy-Drug Sciences, University of Bari, Bari, Italy
Gennaro Agrimi
Department of Biosciences, Biotechnologies and Environment, University of Bari, Bari, Italy
Matteo Ramazzotti
Department of Experimental and Clinical Biomedical Sciences, University of Florence, Florence, Italy
Paola Ostano
Cancer Genomics Lab, Fondazione Edo ed Elvo Tempia, Biella, Italy
Caterina Peraldo Neia
Cancer Genomics Lab, Fondazione Edo ed Elvo Tempia, Biella, Italy
Matteo Parri
Department of Experimental and Clinical Biomedical Sciences, University of Florence, Florence, Italy
Fabrizia Carli
Institute of Clinical Physiology, National Research Council, CNR, Pisa, Italy
Silvia Sabatini
Institute of Clinical Physiology, National Research Council, CNR, Pisa, Italy
Amalia Gastaldelli
Institute of Clinical Physiology, National Research Council, CNR, Pisa, Italy
Fabio Marra
Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy; Corresponding authors. Addresses: Department of Experimental and Clinical Medicine, University of Florence, Cubo Centro Polivalente 2, Viale Pieraccini, 6, I50134 Florence, Italy. Tel.: +39 0552758128 (C. Raggi); Department of Experimental and Clinical Medicine, University of Florence, Largo Brambilla, 3I50134 Florence, Italy. Tel.: +39 0557945425 (F. Marra).
Chiara Raggi
Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy; Corresponding authors. Addresses: Department of Experimental and Clinical Medicine, University of Florence, Cubo Centro Polivalente 2, Viale Pieraccini, 6, I50134 Florence, Italy. Tel.: +39 0552758128 (C. Raggi); Department of Experimental and Clinical Medicine, University of Florence, Largo Brambilla, 3I50134 Florence, Italy. Tel.: +39 0557945425 (F. Marra).
Background & Aims: Among the reprogrammed metabolic pathways described in cancer stem cells, aberrant lipid metabolism has recently drawn increasing attention. Our study explored the contribution of fatty acids (FA) in the regulation of stem-like features in intrahepatic cholangiocarcinoma (iCCA). Methods: We previously identified a functional stem-like subset in human iCCA by using a three-dimensional sphere (SPH) model in comparison to parental cells grown as monolayers (MON). In this study, quantification of intracellular free FA and lipidomic analysis (triacylglycerol [TAG] composition, de novo synthesis products) was performed by Liquid chromatography–mass spectrometry (LC–MS); quadrupole time-of-flight liquid chromatography/mass spectrometry (Q-TOF LC/MS), respectively, in both SPH and MON cultures. Results: Stem-like SPH showed a superior content of free FA (citric, palmitic, stearic, and oleic acids) and unsaturated TAG. Molecularly, SPH showed upregulation of key metabolic enzymes involved in de novo FA biosynthesis (AceCS1, ACLY, ACAC, FASN, ACSL1) and the mTOR signalling pathway. In patients with iCCA (n = 68), tissue expression of FASN, a key gene involved in FA synthesis, correlated with 5-year overall survival. Interference with FASN activity in SPH cells through both specific gene silencing (siRNA) or pharmacological inhibition (orlistat) decreased sphere-forming ability and expression of stem-like markers. In a murine xenograft model obtained by injection of iCCA-SPH cells, FASN inhibition by orlistat or injection of FASN-silenced cells significantly reduced tumour growth and expression of stem-like genes. Conclusion: Altered FA metabolism contributes to the maintenance of a stem-like phenotype in iCCA. FASN inhibition may represent a new approach to interfere with the progression of this deadly disease. Impact and implications: Recent evidence indicates that metabolic disorders correlate with an increased susceptibility to intrahepatic cholangiocarcinoma (iCCA). Our investigation emphasises the pivotal involvement of lipid metabolism in the tumour stem cell biology of iCCA, facilitated by the upregulation of crucial enzymes and the mTOR signalling pathway. From a clinical perspective, this underscores the dual role of FASN as both a prognostic indicator and a therapeutic target, suggesting that FASN inhibitors could enhance patient outcomes by diminishing stemness and tumour aggressiveness. These findings pave the way for novel therapeutic strategies for iCCA and shed light on its relationship with metabolic disorders such as diabetes, obesity, metabolic syndrome, and metabolic dysfunction-associated steatotic liver disease.
