title: The role of disulfide-bridge on the activities of H-shape gemini-like cationic lipid based siRNA delivery.
abstract: In our previous study, a H-shape gemini-like cationic lipid (ssGLCL, formerly named as CLD), composed of two hydrophilic lysine heads and two hydrophobic oleyl alcohol tails with a bridge of the redox-active disulfide-bond, had been synthesized and used as a nanocarrier for delivering small interfering RNAs (siRNAs) into cells. In order to further elucidate the role of disulfide (-S-S-) bridge on the activity of ssGLCL based siRNA delivery, a comparable ccGLCL bridged with a non-reducible carbon-carbon bond was synthesized and used as control in this study. Both two H-shape GLCL molecules could individually self-assemble into cationic nanoparticles in water phase and complex with negatively-charged siRNA into nanoplexes with particle size of ~200nm and zeta potential of ~ +30mV, and exhibit effective siRNA delivery both in vitro and in vivo. Investigation of internalization pathway displayed that both ssGLCL/siRNA and ccGLCL/siRNA nanoplexes were predominantly internalized into MCF-7 cells by the clathrin-mediated endocytosis pattern. Although a lower cellular uptake of siRNA was found in the human breast cancer MCF-7 cells, the ssGLCL/siRNA nanoplexes could exhibit similar or even stronger down-regulation effects on the targeted EGFR mRNA and protein in MCF-7 cells when compared to the ccGLCL/siRNA nanoplexes. Furthermore, mechanistic study showed that the enhanced down-regulation effects of ssGLCL/siRNA nanoplexes on targeted mRNA and protein were probably attributed to the increased release of siRNA from lysosomes to cytoplasm following the cleavage of redox-active disulfide-bridge in ssGLCL. Therefore, we believed that the redox-active H-shape ssGLCL could be a potential nanocarrier towards improving siRNA delivery.
authors.affiliation: State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Beijing, 38 Xueyuan Road, 100191, China.
authors.forename: Xiao-Fei
authors.initials: XF
authors.lastname: Ma
authors.affiliation: State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Beijing, 38 Xueyuan Road, 100191, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Sun
authors.affiliation: State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Beijing, 38 Xueyuan Road, 100191, China.
authors.forename: Chong
authors.initials: C
authors.lastname: Qiu
authors.affiliation: State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Beijing, 38 Xueyuan Road, 100191, China.
authors.forename: Yi-Fan
authors.initials: YF
authors.lastname: Wu
authors.affiliation: State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Beijing, 38 Xueyuan Road, 100191, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Zheng
authors.affiliation: State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Beijing, 38 Xueyuan Road, 100191, China.
authors.forename: Min-Zhi
authors.initials: MZ
authors.lastname: Yu
authors.affiliation: State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Beijing, 38 Xueyuan Road, 100191, China.
authors.forename: Xi-Wei
authors.initials: XW
authors.lastname: Pei
authors.affiliation: State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Beijing, 38 Xueyuan Road, 100191, China.
authors.forename: Lin
authors.initials: L
authors.lastname: Wei
authors.affiliation: State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Beijing, 38 Xueyuan Road, 100191, China.
authors.forename: Yu-Jie
authors.initials: YJ
authors.lastname: Niu
authors.affiliation: State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Beijing, 38 Xueyuan Road, 100191, China.
authors.forename: Wen-Hao
authors.initials: WH
authors.lastname: Pang
authors.affiliation: State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Beijing, 38 Xueyuan Road, 100191, China. Electronic address: yangzj@bjmu.edu.cn.
authors.forename: Zhen-Jun
authors.initials: ZJ
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Beijing, 38 Xueyuan Road, 100191, China. Electronic address: wang-jc@bjmu.edu.cn.
authors.forename: Jian-Cheng
authors.initials: JC
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Beijing, 38 Xueyuan Road, 100191, China.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Zhang
mesh_terms: D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D004220:Disulfides / Q000737:chemistry*; D066246:ErbB Receptors / Q000235:genetics / Q000378:metabolism; D004912:Erythrocytes / Q000187:drug effects; D005260:Female; D020868:Gene Silencing; D018014:Gene Transfer Techniques; D006367:HeLa Cells; D006461:Hemolysis / Q000187:drug effects; D006801:Humans; D008055:Lipids / Q000008:administration & dosage* / Q000737:chemistry; D061986:MCF-7 Cells; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D053758:Nanoparticles / Q000008:administration & dosage* / Q000737:chemistry; D009369:Neoplasms / Q000235:genetics / Q000378:metabolism / Q000628:therapy; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000008:administration & dosage* / Q000737:chemistry; D051381:Rats; D023041:Xenograft Model Antitumor Assays
chemical_list: D004220:Disulfides; D008055:Lipids; D012333:RNA, Messenger; D034741:RNA, Small Interfering; C541485:EGFR protein, mouse; D066246:ErbB Receptors
keywords: Biodegradation; Disulfide bond; Gemini-like lipid; Nanocarrier; siRNA delivery