title: Anti-EphA10 antibody-conjugated pH-sensitive liposomes for specific intracellular delivery of siRNA.
abstract: Therapeutic delivery of small interfering RNA (siRNA) is a major challenge that limits its potential clinical application. Here, a pH-sensitive cholesterol-Schiff base-polyethylene glycol (Chol-SIB-PEG)-modified cationic liposome-siRNA complex, conjugated with the recombinant humanized anti-EphA10 antibody (Eph), was developed as an efficient nonviral siRNA delivery system. Chol-SIB-PEG was successfully synthesized and confirmed with FTIR and (1)H-NMR. An Eph-PEG-SIB-Chol-modified liposome-siRNA complex (EPSLR) was prepared and characterized by size, zeta potential, gel retardation, and encapsulation efficiency. Electrophoresis results showed that EPSLR was resistant to heparin replacement and protected siRNA from fetal bovine serum digestion. EPSLR exhibited only minor cytotoxicity in MCF-7/ADR cells. The results of flow cytometry and confocal laser scanning microscopy suggested that EPSLR enhanced siRNA transfection in MCF-7/ADR cells. Intracellular distribution experiment revealed that EPSLR could escape from the endo-lysosomal organelle and release siRNA into cytoplasm at 4 hours posttransfection. Western blot experiment demonstrated that EPSLR was able to significantly reduce the levels of MDR1 protein in MCF-7/ADR cells. The in vivo study of DIR-labeled complexes in mice bearing MCF-7/ADR tumor indicated that EPSLR could reach the tumor site rather than other organs more effectively. All these results demonstrate that EPSLR has much potential for effective siRNA delivery and may facilitate its therapeutic application.
authors: Department of Pharmaceutics, School of Pharmacy.
authors: Xinlong
authors: X
authors: Zang
authors: Department of Pharmaceutical Chemistry, School of Pharmaceutical Engineering, Shenyang Pharmaceutical University, Shenyang, People's Republic of China.
authors: Huaiwei
authors: H
authors: Ding
authors: Hongqi Hospital affiliated to Mudanjiang Medical University, Mudanjiang, People's Republic of China.
authors: Xiufeng
authors: X
authors: Zhao
authors: Department of Pharmaceutics, School of Pharmacy.
authors: Xiaowei
authors: X
authors: Li
authors: Department of Pharmaceutics, School of Pharmacy.
authors: Zhouqi
authors: Z
authors: Du
authors: Department of Pharmaceutics, School of Pharmacy.
authors: Haiyang
authors: H
authors: Hu
authors: Department of Pharmaceutics, School of Pharmacy.
authors: Mingxi
authors: M
authors: Qiao
authors: Department of Pharmaceutics, School of Pharmacy.
authors: Dawei
authors: D
authors: Chen
authors: Department of Pharmaceutics, School of Pharmacy.
authors: Yuihui
authors: Y
authors: Deng
authors: Department of Pharmaceutics, School of Pharmacy.
authors: Xiuli
authors: X
authors: Zhao
mesh_terms: D000818:Animals; D000906:Antibodies / Q000378:metabolism*; D016923:Cell Death; D002784:Cholesterol / Q000031:analogs & derivatives / Q000138:chemical synthesis / Q000737:chemistry; D024202:Electrophoretic Mobility Shift Assay; D004705:Endocytosis; D005260:Female; D005434:Flow Cytometry; D020868:Gene Silencing; D018014:Gene Transfer Techniques; D006801:Humans; D006863:Hydrogen-Ion Concentration; D042541:Intracellular Space / Q000378:metabolism*; D008081:Liposomes; D061986:MCF-7 Cells; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D008856:Microscopy, Fluorescence; D011092:Polyethylene Glycols / Q000138:chemical synthesis / Q000737:chemistry; D034741:RNA, Small Interfering / Q000008:administration & dosage* / Q000235:genetics; D036081:Receptors, Eph Family / Q000276:immunology*; D012545:Schiff Bases / Q000138:chemical synthesis / Q000737:chemistry
chemical_list: D000906:Antibodies; D008081:Liposomes; D034741:RNA, Small Interfering; D012545:Schiff Bases; C118255:polyethylene glycol-cholesterol; D011092:Polyethylene Glycols; D002784:Cholesterol; C568447:EPHA10 protein, human; D036081:Receptors, Eph Family
keywords: anti-EphA10 antibody; cationic liposome; endosomal escape; gene silencing; pH sensitive; siRNA