Enhanced Delivery of Rose Bengal by Amino Acids Starvation and Exosomes Inhibition in Human Astrocytoma Cells to Potentiate Anticancer Photodynamic Therapy Effects
Bianca Slivinschi,
Federico Manai,
Carolina Martinelli,
Francesca Carriero,
Camilla D’Amato,
Martina Massarotti,
Giorgia Bresciani,
Claudio Casali,
Gloria Milanesi,
Laura Artal,
Lisa Zanoletti,
Federica Milella,
Davide Arfini,
Alberto Azzalin,
Sara Demartis,
Elisabetta Gavini,
Sergio Comincini
Affiliations
Bianca Slivinschi
Department of Biology and Biotechnology, University of Pavia, 27100 Pavia, Italy
Federico Manai
Department of Biology and Biotechnology, University of Pavia, 27100 Pavia, Italy
Carolina Martinelli
Department of Biology and Biotechnology, University of Pavia, 27100 Pavia, Italy
Francesca Carriero
Department of Biology and Biotechnology, University of Pavia, 27100 Pavia, Italy
Camilla D’Amato
Department of Biology and Biotechnology, University of Pavia, 27100 Pavia, Italy
Martina Massarotti
Department of Biology and Biotechnology, University of Pavia, 27100 Pavia, Italy
Giorgia Bresciani
Department of Biology and Biotechnology, University of Pavia, 27100 Pavia, Italy
Claudio Casali
Department of Biology and Biotechnology, University of Pavia, 27100 Pavia, Italy
Gloria Milanesi
Department of Biology and Biotechnology, University of Pavia, 27100 Pavia, Italy
Laura Artal
Department of Biology and Biotechnology, University of Pavia, 27100 Pavia, Italy
Lisa Zanoletti
Department of Biology and Biotechnology, University of Pavia, 27100 Pavia, Italy
Federica Milella
Department of Biology and Biotechnology, University of Pavia, 27100 Pavia, Italy
Davide Arfini
Department of Biology and Biotechnology, University of Pavia, 27100 Pavia, Italy
Alberto Azzalin
Department of Biology and Biotechnology, University of Pavia, 27100 Pavia, Italy
Sara Demartis
Department of Medicine, Surgery and Pharmacy, University of Sassari, 07100 Sassari, Italy
Elisabetta Gavini
Department of Medicine, Surgery and Pharmacy, University of Sassari, 07100 Sassari, Italy
Sergio Comincini
Department of Biology and Biotechnology, University of Pavia, 27100 Pavia, Italy
Photodynamic therapy (PDT) is a promising anticancer strategy based on the light energy stimulation of photosensitizers (PS) molecules within a malignant cell. Among a multitude of recently challenged PS, Rose bengal (RB) has been already reported as an inducer of cytotoxicity in different tumor cells. However, RB displays a low penetration capability across cell membranes. We have therefore developed a short-term amino acids starvation protocol that significantly increases RB uptake in human astrocytoma cells compared to normal rat astrocytes. Following induced starvation uptake, RB is released outside cells by the exocytosis of extracellular vesicles (EVs). Thus, we have introduced a specific pharmacological treatment, based on the GW4869 exosomes inhibitor, to interfere with RB extracellular release. These combined treatments allow significantly reduced nanomolar amounts of administered RB and a decrease in the time interval required for PDT stimulation. The overall conditions affected astrocytoma viability through the activation of apoptotic pathways. In conclusion, we have developed for the first time a combined scheme to simultaneously increase the RB uptake in human astrocytoma cells, reduce the extracellular release of the drug by EVs, and improve the effectiveness of PDT-based treatments. Importantly, this strategy might be a valuable approach to efficiently deliver other PS or chemotherapeutic drugs in tumor cells.