title: Combination of chondroitin sulfate and polyplex micelles from Poly(ethylene glycol)-poly{N'-[N-(2-aminoethyl)-2-aminoethyl]aspartamide} block copolymer for prolonged in vivo gene transfection with reduced toxicity.
abstract: Nonviral polycation-based gene carriers (polyplexes) have attracted attention as safe and efficient gene delivery systems. Polyplex micelles comprised of poly(ethyleneglycol)-block-poly{N'-[N-(2-aminoethyl)-2-aminoethyl]aspartamide} (PEG-PAsp(DET)) and plasmid DNA (pDNA) have shown high transfection efficiency with low toxicity due to the pH-sensitive protonation behavior of PAsp(DET), which enhances endosomal escape, and their self-catalytic degradability under physiological conditions, which reduces cumulative toxicity during transfection. In this study, we improved the safety and transfection efficiency of this polyplex micelle system by adding an anionic polycarbohydrate, chondroitin sulfate (CS). A quantitative assay for cell membrane integrity using image analysis software showed that the addition of CS markedly reduced membrane damage caused by free polycations in the micelle solution. It also reduced tissue damage and subsequent inflammatory responses in the skeletal muscle and lungs of mice following in vivo gene delivery with the polyplex micelles. Subsequently, this led to prolonged transgene expression in the target organs. This combination of polyplex micelles and CS holds great promise for safe and efficient gene introduction in clinical settings.
authors.affiliation: Division of Clinical Biotechnology, Center for Disease Biology and Integrative Medicine, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Uchida
authors.forename: Keiji
authors.initials: K
authors.lastname: Itaka
authors.forename: Qixian
authors.initials: Q
authors.lastname: Chen
authors.forename: Kensuke
authors.initials: K
authors.lastname: Osada
authors.forename: Kanjiro
authors.initials: K
authors.lastname: Miyata
authors.forename: Takehiko
authors.initials: T
authors.lastname: Ishii
authors.forename: Mariko
authors.initials: M
authors.lastname: Harada-Shiba
authors.forename: Kazunori
authors.initials: K
authors.lastname: Kataoka
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D002460:Cell Line; D002462:Cell Membrane / Q000187:drug effects; D002809:Chondroitin Sulfates / Q000737:chemistry* / Q000633:toxicity; D004247:DNA / Q000008:administration & dosage / Q000235:genetics; D004337:Drug Carriers / Q000737:chemistry* / Q000633:toxicity; D005260:Female; D031541:Fluorescence Resonance Energy Transfer; D018014:Gene Transfer Techniques; D049409:Luciferases, Firefly / Q000235:genetics; D008168:Lung / Q000378:metabolism; D051379:Mice; D008813:Mice, Inbred ICR; D008823:Micelles; D018482:Muscle, Skeletal / Q000378:metabolism; D011092:Polyethylene Glycols / Q000737:chemistry* / Q000633:toxicity; D011506:Proteins / Q000737:chemistry* / Q000633:toxicity; D013050:Spectrometry, Fluorescence; D013499:Surface Properties; D013997:Time Factors; D014162:Transfection
chemical_list: D004337:Drug Carriers; D008823:Micelles; D011506:Proteins; C571441:poly(ethylene glycol)-poly(N'-(N-(2-aminoethyl)-2-aminoethyl)aspartamide) block copolymer; D011092:Polyethylene Glycols; D002809:Chondroitin Sulfates; D004247:DNA; D049409:Luciferases, Firefly