title: Effect of anionic PEGylated polypeptide on gene transfection mediated by glycolipid conjugate micelles.
abstract: To improve the gene transfection efficiency mediated by chitosan-g-stearic acid (CS) micelles, poly(ethylene glycol)-b-poly(γ-glutamic acid) (PG) was incorporated into a CS-based gene delivery system. CS/PG/pDNA complexes were prepared by ionic interaction. CS and PEGylated CS (PCS) micelles were introduced to prepare binary complexes for use as controls. CS/PG/pDNA complexes possessed similar sizes and presented as irregular spheroids in shape. The incorporation of PG into CS/pDNA complexes did not affect the ability of CS to compact pDNA and also showed a protective effect against DNase I based degradation of pDNA. Importantly, PG could increase gene transfection efficiency, which was also affected by the mixing methods used for the preparation of CS/PG/pDNA ternary complexes. The transfection efficiencies mediated by CS/PG/pDNA complexes against HEK293 and EC-1 cells reached up to 40.8% and 11.6%, respectively, which were much higher than those of CS/pDNA complexes (1.3% and 4.0%) and PCS/pDNA complexes (0.8% and 2.4%). In addition, the incorporation of PG into CS/pDNA complexes significantly enhanced cellular uptake in HEK293 and EC-1 cells and, additionally, improved endosomal escape and intracellular vector unpacking. However, the incorporation of PG reduced the cellular uptake of CS/PG/pDNA complexes in macrophages (RAW264.7 cells). It was further demonstrated that, in addition to a nonspecific charge-mediated binding to cell membranes, a γ-PGA-specific receptor-mediated pathway was involved in the internalization of CS/PG/pDNA complexes. These results indicated that PG played multiple important roles in enhancing the transfection efficiency of CS/PG/pDNA complexes.
authors.affiliation: College of Pharmaceutical Sciences, Zhejiang University, 866 Yuhangtang Road, Hangzhou 310058, People's Republic of China.
authors.forename: Han-Xi
authors.initials: HX
authors.lastname: Yi
authors.affiliation: College of Pharmaceutical Sciences, Zhejiang University, 866 Yuhangtang Road, Hangzhou 310058, People's Republic of China.
authors.forename: Jie
authors.initials: J
authors.lastname: Wu
authors.affiliation: College of Pharmaceutical Sciences, Zhejiang University, 866 Yuhangtang Road, Hangzhou 310058, People's Republic of China.
authors.forename: Yong-Zhong
authors.initials: YZ
authors.lastname: Du
authors.affiliation: College of Pharmaceutical Sciences, Zhejiang University, 866 Yuhangtang Road, Hangzhou 310058, People's Republic of China.
authors.forename: Ying-Wen
authors.initials: YW
authors.lastname: Hu
authors.affiliation: College of Pharmaceutical Sciences, Zhejiang University, 866 Yuhangtang Road, Hangzhou 310058, People's Republic of China.
authors.forename: Hong
authors.initials: H
authors.lastname: Yuan
authors.affiliation: College of Pharmaceutical Sciences, Zhejiang University, 866 Yuhangtang Road, Hangzhou 310058, People's Republic of China.
authors.forename: Jian
authors.initials: J
authors.lastname: You
authors.affiliation: College of Pharmaceutical Sciences, Zhejiang University, 866 Yuhangtang Road, Hangzhou 310058, People's Republic of China.
authors.forename: Fu-Qiang
authors.initials: FQ
authors.lastname: Hu
mesh_terms: D000818:Animals; D000838:Anions; D048271:Chitosan / Q000737:chemistry; D004247:DNA / Q000737:chemistry; D004587:Electrophoresis, Agar Gel; D011992:Endosomes / Q000378:metabolism; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D006017:Glycolipids / Q000737:chemistry*; D057809:HEK293 Cells; D006801:Humans; D008264:Macrophages / Q000378:metabolism; D051379:Mice; D008823:Micelles; D018613:Microscopy, Confocal; D046529:Microscopy, Electron, Transmission; D010455:Peptides / Q000737:chemistry*; D011092:Polyethylene Glycols / Q000737:chemistry*; D011099:Polyglutamic Acid / Q000031:analogs & derivatives / Q000737:chemistry; D011108:Polymers / Q000737:chemistry; D013229:Stearic Acids / Q000737:chemistry; D014162:Transfection
chemical_list: D000838:Anions; D006017:Glycolipids; D008823:Micelles; D010455:Peptides; D011108:Polymers; D013229:Stearic Acids; C511775:poly(gamma-glutamic acid); D011099:Polyglutamic Acid; D011092:Polyethylene Glycols; C031183:stearic acid; D004247:DNA; D048271:Chitosan
keywords: cellular uptake; chitosan-g-stearic acid; endosomal escape; poly(ethylene glycol)-b-poly(γ-glutamic acid); transfection efficiency; vector unpacking