Selective targeting of melanoma by PEG-masked protein-based multifunctional nanoparticles

Abstract

Luca Vannucci1,*, Elisabetta Falvo2,*, Manuela Fornara3, Patrizio Di Micco4, Oldrich Benada1, Jiri Krizan1, Jan Svoboda1, Katarina Hulikova-Capkova1, Veronica Morea3, Alberto Boffi4,5, Pierpaolo Ceci3 1Institute of Microbiology, Academy of Sciences of the Czech Republic, VVI, Prague, Czech Republic; 2Regina Elena Cancer Institute, Pharmacokinetic/Pharmacogenomic Unit, 3National Research Council of Italy, Institute of Molecular Biology and Pathology, 4Department of Biochemical Sciences “A Rossi Fanelli”, University of Rome “Sapienza”, 5Center for Life Nano Science at Sapienza, Italian Institute of Technology, Rome, Italy *These two authors contributed equally to this workBackground: Nanoparticle-based systems are promising for the development of imaging and therapeutic agents. The main advantage of nanoparticles over traditional systems lies in the possibility of loading multiple functionalities onto a single molecule, which are useful for therapeutic and/or diagnostic purposes. These functionalities include targeting moieties which are able to recognize receptors overexpressed by specific cells and tissues. However, targeted delivery of nanoparticles requires an accurate system design. We present here a rationally designed, genetically engineered, and chemically modified protein-based nanoplatform for cell/tissue-specific targeting.Methods: Our nanoparticle constructs were based on the heavy chain of the human protein ferritin (HFt), a highly symmetrical assembly of 24 subunits enclosing a hollow cavity. HFt-based nanoparticles were produced using both genetic engineering and chemical functionalization methods to impart several functionalities, ie, the α-melanocyte-stimulating hormone peptide as a melanoma-targeting moiety, stabilizing and HFt-masking polyethylene glycol molecules, rhodamine fluorophores, and magnetic resonance imaging agents. The constructs produced were extensively characterized by a number of physicochemical techniques, and assayed for selective melanoma-targeting in vitro and in vivo.Results: Our HFt-based nanoparticle constructs functionalized with the α-melanocyte-stimulating hormone peptide moiety and polyethylene glycol molecules were specifically taken up by melanoma cells but not by other cancer cell types in vitro. Moreover, experiments in melanoma-bearing mice indicate that these constructs have an excellent tumor-targeting profile and a long circulation time in vivo.Conclusion: By masking human HFt with polyethylene glycol and targeting it with an α-melanocyte-stimulating hormone peptide, we developed an HFt-based melanoma-targeting nanoplatform for application in melanoma diagnosis and treatment. These results could be of general interest, because the same strategy can be exploited to develop ad hoc nanoplatforms for specific delivery towards any cell/tissue type for which a suitable targeting moiety is available.Keywords: multifunctional nanoparticles, ferritin, nanoplatform, cancer-targeting, melanoma