title: Binary and ternary complexes based on polycaprolactone-graft-poly (N, N-dimethylaminoethyl methacrylate) for targeted siRNA delivery.
abstract: Small interfering RNA (siRNA) is a powerful gene silencing tool and has promising prospects in basic research and the development of therapeutic reagents. However, the lack of an effective and safe tool for siRNA delivery hampers its application. Here, we introduced binary and ternary complexes that effectively mediated siRNA-targeted gene silencing. Both complexes showed excellent siRNA loading even at the low N/P/C ratio of 3:1:0. FACS and confocal microscopy demonstrated that nearly all cells robustly internalized siRNAs into the cytoplasm, where RNA interference (RNAi) occurred. Luciferase assay and Western blot verified that silencing efficacy reached >80%, and introducing folate onto the ternary complexes further enhanced silencing efficacy by about 10% over those without folate at the same N/P/C ratio. In addition, the coating of PGA-g-mPEG decreased the zeta potential almost to electroneutrality, and the MTT assay showed decreased cytotoxicity. In vivo distribution measurement and histochemical analysis executed in C57BL/6 and Hela tumor-bearing BALB/c nude mice showed that complexes accumulated in the liver, lungs, pancreas and tumors and were released slowly for a long time after intravenous injection. Furthermore, ternary complexes showed higher siRNA fluorescence intensity than binary complexes at the same N/P ratio in tumor tissues, those with folate delivered more siRNAs to tumors than those without folate, and more folate induced more siRNA transport to tumors. In addition, in vivo functional study showed that both binary and ternary complexes mediated down-regulation of ApoB in liver efficiently and consequently blocked the secretion of fatty acids into the blood, resulted in lipid accumulation in liver, liver steatosis and hepatic dysfunction. In conclusion, these complexes provided a powerful means of administration for siRNA-mediated treatment of liver-related diseases and various cancers, especial for pancreatic and cervical cancer.
authors.affiliation: Laboratory of Nucleic Acid Technology, Institute of Molecular Medicine, Peking University, Beijing 100871, China.
authors.forename: Yuanyu
authors.initials: Y
authors.lastname: Huang
authors.forename: Daoshu
authors.initials: D
authors.lastname: Lin
authors.forename: Qian
authors.initials: Q
authors.lastname: Jiang
authors.forename: Wendi
authors.initials: W
authors.lastname: Zhang
authors.forename: Shutao
authors.initials: S
authors.lastname: Guo
authors.forename: Ping
authors.initials: P
authors.lastname: Xiao
authors.forename: Shuquan
authors.initials: S
authors.lastname: Zheng
authors.forename: Xiaoxia
authors.initials: X
authors.lastname: Wang
authors.forename: Hongbo
authors.initials: H
authors.lastname: Chen
authors.forename: Hong-Yan
authors.initials: HY
authors.lastname: Zhang
authors.forename: Liandong
authors.initials: L
authors.lastname: Deng
authors.forename: Jinfeng
authors.initials: J
authors.lastname: Xing
authors.forename: Quan
authors.initials: Q
authors.lastname: Du
authors.forename: Anjie
authors.initials: A
authors.lastname: Dong
authors.forename: Zicai
authors.initials: Z
authors.lastname: Liang
mesh_terms: D000818:Animals; D015153:Blotting, Western; D005260:Female; D005434:Flow Cytometry; D005822:Genetic Vectors / Q000008:administration & dosage* / Q000737:chemistry*; D006801:Humans; D008768:Methylmethacrylates / Q000737:chemistry*; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008819:Mice, Nude; D011091:Polyesters / Q000737:chemistry*; D011108:Polymers / Q000737:chemistry*; D034741:RNA, Small Interfering / Q000008:administration & dosage* / Q000737:chemistry*; D023041:Xenograft Model Antitumor Assays
chemical_list: D008768:Methylmethacrylates; D011091:Polyesters; D011108:Polymers; D034741:RNA, Small Interfering; C016240:polycaprolactone