title: What the cell surface does not see: The gene vector under the protein corona.
abstract: The fate of lipid-based nanovectors, used in genetic targeting inside cells, depends on their behavior in biological media. In fact, during both in vitro and in vivo transfection, nanovectors come in contact with proteins that compete for their surface and build the protein corona, their true biological identity while engaging the cell membrane. Nonetheless, after cell internalization, the efficacy of transfection may depend also on structural modifications that occurred under the protein cover, following interaction with biological fluids. Here, based on previous in vivo experiments, two widely used lipid mixtures, namely DOTAP/DOPC and DC-Chol/DOPE, were identified as paradigms to investigate the impact of the inner structure of nanovectors on the transfection efficiency, all being proficiently internalized. The evolution of the inner structure of cationic lipoplexes and nanoparticles based on such lipid mixtures, following interaction with human plasma, could be unraveled. Particles were investigated in high dilution, approaching the biosimilar conditions. Data have demonstrated that the modulation of their inner structure depends on their lipid composition and the plasma concentration, still preserving the genetic payload. Interestingly, protein contact induces a variety of inner structures with different perviousness, including reshaping into cubic phases of different porosity, sometimes observed upon interaction between carrier-lipids and cell-lipids. Cubic reshaping is of biological relevance, as lipid cubic phases have been recently associated to both fusogenicity and to the readiness in releasing the payload to the final target via endosomal escape.
authors: Dept. Medical Biotechnologies and Translational Medicine, University of Milano, LITA, Via F.lli Cervi, 93, 20090, Segrate, Milano, Italy.
authors: Simona
authors: S
authors: Motta
authors: Dept. Medical Biotechnologies and Translational Medicine, University of Milano, LITA, Via F.lli Cervi, 93, 20090, Segrate, Milano, Italy.
authors: Valeria
authors: V
authors: Rondelli
authors: Dept. Medical Biotechnologies and Translational Medicine, University of Milano, LITA, Via F.lli Cervi, 93, 20090, Segrate, Milano, Italy.
authors: Laura
authors: L
authors: Cantu
authors: Dept. Medical Biotechnologies and Translational Medicine, University of Milano, LITA, Via F.lli Cervi, 93, 20090, Segrate, Milano, Italy.
authors: Elena
authors: E
authors: Del Favero
authors: Dept. Medical Biotechnologies and Translational Medicine, University of Milano, LITA, Via F.lli Cervi, 93, 20090, Segrate, Milano, Italy.
authors: Massimo
authors: M
authors: Aureli
authors: Department of Molecular Medicine, Sapienza University of Rome, Viale Regina Elena 261, 00185 Rome, Italy.
authors: Daniela
authors: D
authors: Pozzi
authors: Department of Molecular Medicine, Sapienza University of Rome, Viale Regina Elena 261, 00185 Rome, Italy.
authors: Giulio
authors: G
authors: Caracciolo
authors: Dept. Medical Biotechnologies and Translational Medicine, University of Milano, LITA, Via F.lli Cervi, 93, 20090, Segrate, Milano, Italy. Electronic address: paola.brocca@unimi.it.
authors: Paola
authors: P
authors: Brocca
mesh_terms: D002412:Cations / Q000737:chemistry; D002462:Cell Membrane / Q000737:chemistry* / Q000378:metabolism; D002784:Cholesterol / Q000031:analogs & derivatives / Q000737:chemistry; D004247:DNA / Q000737:chemistry* / Q000235:genetics; D005229:Fatty Acids, Monounsaturated / Q000737:chemistry; D005822:Genetic Vectors / Q000737:chemistry* / Q000235:genetics; D006801:Humans; D008055:Lipids / Q000737:chemistry; D008081:Liposomes / Q000737:chemistry; D053758:Nanoparticles / Q000737:chemistry*; D010713:Phosphatidylcholines / Q000737:chemistry; D010714:Phosphatidylethanolamines / Q000737:chemistry; D010949:Plasma; D000066970:Protein Corona / Q000737:chemistry*; D000644:Quaternary Ammonium Compounds / Q000737:chemistry; D053838:Scattering, Small Angle; D014162:Transfection / Q000379:methods*; D014961:X-Ray Diffraction
chemical_list: D002412:Cations; D005229:Fatty Acids, Monounsaturated; D008055:Lipids; D008081:Liposomes; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D000066970:Protein Corona; D000644:Quaternary Ammonium Compounds; C070648:3-(N-(N',N'-dimethylaminoethane)carbamoyl)cholesterol; C094877:dioleoyl phosphatidylethanolamine; D004247:DNA; D002784:Cholesterol; C017251:1,2-oleoylphosphatidylcholine; C070046:1,2-dioleoyloxy-3-(trimethylammonium)propane
keywords: Cubic phases; Gene delivery; Lipoplex plasma interaction; Protein corona; Protein–lipid interaction; X-ray spectroscopy