title: Enzyme-responsive destabilization of stabilized plasmid-lipid nanoparticles as an efficient gene delivery.
abstract: Stabilized plasmid-lipid particles (SPLPs) have been developed to overcome the low stability issue of cationic liposomes, however, SPLPs that are too stable result in unsatisfactory transfection efficiency. In this article, we prepared enzyme-responsive SPLPs (eSPLPs) composed of 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), and mPEG-GLFG-K-(C16)2, a PEG lipid with an enzymatically-cleavable linker (glycine-phenylalanine-leucine-glycine, GFLG). eSPLPs were successfully prepared with plasmid DNA (pDNA) encapsulation efficiency of over 80%, using the detergent dialysis method. The PEG shell stabilized eSPLPs and maintained a hydrodynamic diameter of around 200nm. Although typical SPLPs were relatively intact in endosomal condition, the PEG shell of eSPLPs was cleaved following the degradation of the GFLG linker by cathepsin B in the endosome. Then, eSPLPs collapsed and induced endosomal disruption triggering the controlled release of the encapsulated pDNA into cytoplasm. Owing to the enzyme-responsive destabilization, eSPLPs showed a 10 to 100-fold higher transfection efficiency than control SPLPs, which was confirmed using luciferase assay. These results suggest that eSPLPs might be promising candidates for practical use as gene delivery systems, with both stability and high transfection efficiency for future in vivo applications.
authors.affiliation: Department of Biochemistry, Chungnam National University, Daejeon 305-764, Republic of Korea.
authors.forename: Su Jeong
authors.initials: SJ
authors.lastname: Song
authors.affiliation: Department of Biochemistry, Chungnam National University, Daejeon 305-764, Republic of Korea.
authors.forename: Seulgi
authors.initials: S
authors.lastname: Lee
authors.affiliation: Department of Chemistry, Seoul National University, Seoul 157-742, Republic of Korea. Electronic address: gacn@snu.ac.kr.
authors.forename: Yan
authors.initials: Y
authors.lastname: Lee
authors.affiliation: Department of Biochemistry, Chungnam National University, Daejeon 305-764, Republic of Korea. Electronic address: joonsig@cnu.ac.kr.
authors.forename: Joon Sig
authors.initials: JS
authors.lastname: Choi
mesh_terms: D002401:Cathepsin B / Q000737:chemistry; D002470:Cell Survival / Q000187:drug effects; D004247:DNA / Q000737:chemistry; D005229:Fatty Acids, Monounsaturated / Q000737:chemistry; D018014:Gene Transfer Techniques; D057809:HEK293 Cells; D006801:Humans; D053758:Nanoparticles / Q000008:administration & dosage* / Q000737:chemistry; D009842:Oligopeptides / Q000737:chemistry; D010714:Phosphatidylethanolamines / Q000737:chemistry; D010957:Plasmids; D011092:Polyethylene Glycols / Q000737:chemistry; D000644:Quaternary Ammonium Compounds / Q000737:chemistry
chemical_list: D005229:Fatty Acids, Monounsaturated; D009842:Oligopeptides; D010714:Phosphatidylethanolamines; D000644:Quaternary Ammonium Compounds; D011092:Polyethylene Glycols; C020888:1,2-dielaidoylphosphatidylethanolamine; D004247:DNA; D002401:Cathepsin B; C070046:1,2-dioleoyloxy-3-(trimethylammonium)propane
keywords: Cathepsin B; Enzyme-responsive nanoparticles; GFLG; Gene delivery; Liposome; PEGylation; Stabilized plasmid-lipid particles (SPLPs)