title: Improved Doxorubicin Encapsulation and Pharmacokinetics of Ferritin-Fusion Protein Nanocarriers Bearing Proline, Serine, and Alanine Elements.
abstract: A novel human ferritin-based nanocarrier, composed of 24 modified monomers able to auto-assemble into a modified protein cage, was produced and used as selective carrier of anti-tumor payloads. Each modified monomer derives from the genetic fusion of two distinct modules, namely the heavy chain of human ferritin (HFt) and a stabilizing/protective PAS polypeptide sequence rich in proline (P), serine (S), and alanine (A) residues. Two genetically fused protein constructs containing PAS polymers with 40- and 75-residue lengths, respectively, were compared. They were produced and purified as recombinant proteins in Escherichia coli at high yields. Both preparations were highly soluble and stable in vitro as well as in mouse plasma. Size-exclusion chromatography, dynamic light scattering, and transmission electron microscopy results indicated that PASylated ferritins are fully assembled and highly monodispersed. In addition, yields and stability of encapsulated doxorubicin were significantly better for both HFt-PAS proteins than for wild-type HFt. Importantly, PAS sequences considerably prolonged the half-life of HFt in the mouse bloodstream. Finally, our doxorubicin-loaded nanocages preserved the pharmacological activity of the drug. Taken together, these results indicate that both of the developed HFt-PAS fusion proteins are promising nanocarriers for future applications in cancer therapy.
authors.affiliation: Institute of Molecular Biology and Pathology CNR, National Research Council of Italy , 00185 Rome, Italy.|Department of Biochemical Sciences "A. Rossi Fanelli", University "Sapienza" , 00185 Rome, Italy.
authors.forename: Elisabetta
authors.initials: E
authors.lastname: Falvo
authors.affiliation: Regina Elena National Cancer Institute , 00144 Rome, Italy.
authors.forename: Elisa
authors.initials: E
authors.lastname: Tremante
authors.affiliation: Istituto di Biochimica e Biochimica Clinica, Università Cattolica del Sacro Cuore , Largo F. Vito 1, 00168 Rome, Italy.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Arcovito
authors.affiliation: Istituto di Fisica, Università Cattolica del Sacro Cuore , Largo F. Vito 1, 00168, Rome, Italy.
authors.forename: Massimiliano
authors.initials: M
authors.lastname: Papi
authors.affiliation: Department of Chemical Research Support, Weizmann Institute of Science , Rehovot 7610001, Israel.
authors.forename: Nadav
authors.initials: N
authors.lastname: Elad
authors.affiliation: Institute of Molecular Biology and Pathology CNR, National Research Council of Italy , 00185 Rome, Italy.|Department of Biochemical Sciences "A. Rossi Fanelli", University "Sapienza" , 00185 Rome, Italy.|Center for Life Nano Science at Sapienza, Istituto Italiano di Tecnologia (IIT) , 00161 Rome, Italy.
authors.forename: Alberto
authors.initials: A
authors.lastname: Boffi
authors.affiliation: Institute of Molecular Biology and Pathology CNR, National Research Council of Italy , 00185 Rome, Italy.
authors.forename: Veronica
authors.initials: V
authors.lastname: Morea
authors.affiliation: Department of Neurological, Biomedical and Movement Sciences, University of Verona , 37134 Verona, Italy.
authors.forename: Giamaica
authors.initials: G
authors.lastname: Conti
authors.affiliation: Experimental and Clinical Pharmacology Unit, CRO Aviano National Cancer Institute , 33081Aviano (Pordenone), Italy.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Toffoli
authors.affiliation: Department of Medicine, University of Verona , 37134 Verona, Italy.
authors.forename: Giulio
authors.initials: G
authors.lastname: Fracasso
authors.affiliation: Regina Elena National Cancer Institute , 00144 Rome, Italy.
authors.forename: Patrizio
authors.initials: P
authors.lastname: Giacomini
authors.affiliation: Institute of Molecular Biology and Pathology CNR, National Research Council of Italy , 00185 Rome, Italy.
authors.forename: Pierpaolo
authors.initials: P
authors.lastname: Ceci
mesh_terms: D000409:Alanine / Q000737:chemistry; D000818:Animals; D000903:Antibiotics, Antineoplastic / Q000737:chemistry* / Q000493:pharmacokinetics / Q000494:pharmacology; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D004317:Doxorubicin / Q000737:chemistry* / Q000493:pharmacokinetics / Q000494:pharmacology; D004354:Drug Screening Assays, Antitumor; D004355:Drug Stability; D005293:Ferritins / Q000737:chemistry; D006207:Half-Life; D006801:Humans; D008807:Mice, Inbred BALB C; D053769:Nanocapsules / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry; D011092:Polyethylene Glycols / Q000737:chemistry; D011392:Proline / Q000737:chemistry; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D012694:Serine / Q000737:chemistry
chemical_list: D000903:Antibiotics, Antineoplastic; D053769:Nanocapsules; D010455:Peptides; D011993:Recombinant Fusion Proteins; D011092:Polyethylene Glycols; D012694:Serine; D004317:Doxorubicin; D005293:Ferritins; D011392:Proline; D000409:Alanine