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direct cytosolic siRNA delivery by reconstituted high density lipoprotein for target-specific therapy of tumor angiogenesis

Direct cytosolic siRNA delivery by reconstituted high densit...

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第十二届全国青年药学工作者最新科研成果交流会

作者： Yang Ding Yazhe Wang Wei Wang Xiaochen Gu Jianping Zhou Qiang Zhang State Key Laboratory of Natural Medicines Department of PharmaceuticsChina Pharmaceutical University Faculty of Pharmacy University of Manitoba750 McDermot AvenueWinnipegMBCanada R3E 0T5 State Key Laboratory of Natural and Biomimetic Drugs School of Pharmaceutical SciencesPeking University

We described here the mechanisms by which small interfering RNA（siRNA） molecules incorporated in reconstituted high density lipoprotein（rHDL） were efficiently transferred into the cytoplasm of cells to perform target-specific therapy of tumor *** fluorescent-tagged apolipoprotein A-I（apoA-1） and cholesterol-conjugated siRNA（Chol-siRNA）,it was confirmed with FACS and confocal microscopic measurements that Chol-siRNA-loaded rHDL nanoparticles（rHDL/Chol-siRNA complexes） were successfully established and apoA-I certainly was attached to the surface of Chol-siRNA-loaded lipoplexes（Lipos/Chol-siRNA complexes）.Stably assembled rHDL/Chol-siRNA complexes demonstrated proper nanosize,quasi-spherical shape and improved nuclease protection over naked *** was also interesting to note that rHDL provided a highly effective approach to transfer Chol-siRNA across the membrane directly into the cytoplasm via the scavenger receptor BI（SR-BI）-mediated non-endocytotic mechanism,thereby bypassing endo-lysosomal *** also showed clear evidence that the in vitro implementation of rHDL for Chol-siRNA-VEGF（Chol-siRNA targeting vascular endothelial growth factor gene） delivery markedly promoted RNA interference（RNAi）-mediated degradation of VEGF mRNA,resulting in down-regulation of secreted VEGF *** vivo fluorescence imaging indicated that near-infrared（MR） dye Cy5 labeled Chol-siRNA-loaded rHDL nanoparticles（rHDL/Cy5-Chol-siRNA complexes） displayed long circulation time,SR-BI positive tumor-selective targeting,and efficient cytosolic delivery ***,intravenous administration of Chol-siRNA-VEGF-loaded rHDL nanoparticles（rHDL/Chol-siRNA-VEGF complexes） significantly enhanced anti-tumor efficacy against breast cancer,decreased VEGF expression level,and inhibited formation of intratumoral microvessels at the tumor *** was concluded that rHDL possessed therapeutic potential and versatility in mediating Chol-siRNA-VEGF direct cytosoli

关键词： Reconstituted high density lipoprotein Cholesterol-conjugated siRNA direct cytosolic delivery Vascular endothelial growth factor Tumor-selective accumulation Anti-angiogenic therapy

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