Novel serum-tolerant lipoplexes target the folate receptor efficiently

作者：Gorle Sridevi; Ariatti Mario^*; Singh Moganavelli

来源：European Journal of Pharmaceutical Sciences, 2014, 59: 83-93.

DOI：10.1016/j.ejps.2014.04.012

• Gene therapy
• Non-viral vectors
• Lipid nanoparticles
• Liposome
• Folate receptor
• Transfection

Gene transfer using non-viral vectors is a promising approach for the safe delivery of nucleic acid therapeutics. In this study, we investigate a lipid-based system for targeted gene delivery to malignant cells overexpressing the folate receptor (FR). Cationic liposomes were formulated with and without the targeting ligand folate conjugated to distearoylphosphatidyl ethanolamine polyethylene glycol 2000 (DSPE-PEG(2000)), the novel cytofectin 3 beta[N(N-1,N-1-dimethlaminopropylsuccinamidoethane)-carbamoyl]cholesterol (SG04), which contains a 13 atom, 15 angstrom spacer element, and the helper lipid, dioleoylphosphatidylethanolamine (DOPE). Physicochemical parameters of the liposomes and lipoplexes were obtained by zeta sizing, zeta potential measurement and cryo-TEM. DNA-binding and protection capabilities of liposomes were confirmed by gel retardation assays, EtBr intercalation and nuclease protection assays. The complexes were assessed in an in vitro system for their effect on cell viability using the MIT assay, and gene transfection activity using the luciferase assay in three cell lines; HEK293 (FR-negative), HeLa (FR+-positive), KB (FR++-positive). Low cytotoxicities were observed in all cell lines, while transgene activity promoted by folate-tagged lipoplexes in FR-positive lines was tenfold greater than that by untargeted constructs and cell entry by folate complexes was demonstrably by FR mediation. These liposome formulations have the design capacity for in vivo application and may therefore be promising candidates for further development.

• 出版日期2014-8-1