Anti-EphA10 antibody-conjugated pH-sensitive liposomes for specific intracellular delivery of siRNA.
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.
['Zang|Xinlong|X|', 'Ding|Huaiwei|H|', 'Zhao|Xiufeng|X|', 'Li|Xiaowei|X|', 'Du|Zhouqi|Z|', 'Hu|Haiyang|H|', 'Qiao|Mingxi|M|', 'Chen|Dawei|D|', 'Deng|Yuihui|Y|', 'Zhao|Xiuli|X|']
D000818:Animals D000906:Antibodies D016923:Cell Death D002784:Cholesterol 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 D008081:Liposomes D061986:MCF-7 Cells D051379:Mice D008807:Mice, Inbred BALB C D008819:Mice, Nude D008856:Microscopy, Fluorescence D011092:Polyethylene Glycols D034741:RNA, Small Interfering D036081:Receptors, Eph Family D012545:Schiff Bases
siRNA anti-EphA10 antibody cationic liposome endosomal escape gene silencing pH sensitive