International Journal of Nanomedicine (Jan 2019)
Pemetrexed-loaded nanoparticles targeted to malignant pleural mesothelioma cells: an in vitro study
Abstract
Emanuela Cova,1 Laura Pandolfi,1 Miriam Colombo,2 Vanessa Frangipane,1 Simona Inghilleri,1 Monica Morosini,1 Simona Mrakic-Sposta,3 Sarah Moretti,3 Manuela Monti,4 Ymera Pignochino,5 Silvia Benvenuti,5 Davide Prosperi,2,6 Giulia Stella,1 Patrizia Morbini,7 Federica Meloni8 1Clinic of Lung Diseases, IRCCS Foundation Policlinico San Matteo, Pavia, Italy; 2Deparment of Biotechnology and Bioscience, University of Milano - Bicocca, Milan, Italy; 3National Council of Research, Institute of Bioimaging and Molecular Physiology, Segrate, Milan, Italy; 4Laboratory of Biotechnology, Research Center of Rigenerative Medicine, IRCCS Foundation Policlinico San Matteo, Pavia, Italy; 5Experimental Clinical Molecular Oncology, IRCCS Candiolo Cancer Institute-FPO, Candiolo, Turin, Italy; 6Laboratory of Nanomedicine, Clinical Institute of Maugeri, S.p.A., Pavia, Italy; 7Department of Molecular Medicine, Pathology Unit, IRCCS Foundation Policlinico San Matteo, Pavia, Italy; 8Department of Internal Medicine, Pneumology Unit, University of Pavia, Pavia, Italy Purpose: Malignant pleural mesothelioma (MPM) is an aggressive tumor characterized by poor prognosis. Its incidence is steadily increasing due to widespread asbestos exposure. There is still no effective therapy for MPM. Pemetrexed (Pe) is one of the few chemotherapeutic agents approved for advanced-stage disease, although the objective response to the drug is limited. The use of gold nanoparticles (GNPs) as a drug delivery system promises several advantages, including specific targeting of malignant cells, with increased intracellular drug accumulation and reduced systemic toxicity, and, in the case of MPM, direct treatment administration into the pleural space. This study aims at exploring CD146 as a potential MPM cell-specific target for engineered Pe-loaded GNPs and to assess their effectiveness in inhibiting MPM cell line growth.Methods: MPM cell lines and primary cultures obtained by pleural effusions from MPM patients were assayed for CD146 expression by flow cytometry. Internalization by MPM cell lines of fluorescent dye-marked GNPs decorated with a monoclonal anti CD146 coated GNPs (GNP-HC) was proven by confocal microscopy. The effects of anti CD146 coated GNPs loaded with Pe (GNP-HCPe) on MPM cell lines were evaluated by cell cycle (flow cytometry), viability (MTT test), clonogenic capacity (soft agar assay), ROS production (electric paramagnetic resonance), motility (wound healing assay), and apoptosis (flow cytometry).Results: GNP-HC were selectively uptaken by MPM cells within 1 hour. MPM cell lines were blocked in the S cell cycle phase in the presence of GNP-HCPe. Both cell viability and motility were significantly affected by nanoparticle treatment compared to Pe. Apoptotic rate and ROS production were significantly higher in the presence of nanoparticles. Clonogenic capacity was completely inhibited following nanoparticle internalization.Conclusion: GNP-HCPe treatment displays in vitro antineoplastic action and is more effective than Pe alone in inhibiting MPM cell line malignant phenotype. The innovative use of specifically targeted GNPs opens the perspective of local intrapleural administration to avoid normal cell toxicity and enhance chemotherapy efficacy. Keywords: gold nanoparticles, nanodrug delivery, mesothelioma, pemetrexed, intrapleural delivery
