Targeted delivery of siRNA to hepatocytes and hepatic stellate cells by bioconjugation.
Targeted delivery of siRNA to hepatocytes and hepatic stellate cells by bioconjugation.
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DOI：
10.1021/bc100346n
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发表时间：
2010-11-17
影响因子：
4.7
通讯作者：
Mahato, Ram I.
中科院分区：
文献类型：
--
作者：
Zhu, Lin;Mahato, Ram I.
以前，我们成功地通过不稳定的酸酯连接物将半乳糖化聚乙二醇(Gal-PEG)连接到寡核苷酸(ODN)(朱等人，Biocongig Chem，2008，19：290-8)。在本研究中，将反义siRNA链连接到Gal-PEG和甘露糖6-磷酸聚乙二醇(M6P-PEG)上，分别向肝细胞和肝星状细胞(HSCs)靶向传递siRNA。用离子交换层析对这些siRNA偶联物进行了纯化，并用凝胶滞留实验进行了验证。为了评价它们的RNA干扰功能，将针对萤火虫荧光素酶和转化生长因子β1(TGF1)基因的小干扰RNA双链连接到Gal-β和M6P-Eg上，并检测了它们的基因沉默效率。用二硫苏糖醇裂解聚乙二醇二硫键，释放完整的siRNA。Gal-PEG-siRNA和M6P-PEG-siRNA结合物在不加任何转染剂的情况下均可使荧光素酶基因表达沉默约40%，而相同剂量的阳离子脂质体对荧光素酶基因的沉默效果可达98%以上。Gal-β和M6P-PEG与转化生长因子-β-1siRNA偶联后，也能抑制内源性转化生长因子-CD11siRNA的表达。综上所述，这些siRNA偶联物有可能将siRNA靶向输送到肝细胞和肝星状细胞，从而在体内有效地进行基因沉默。
Previously, we successfully conjugated galactosylated poly(ethylene glycol) (Gal-PEG) to oligonucleotides (ODNs) via an acid labile ester linker (Zhu et al., Bioconjug Chem, 2008, 19: 290-8). In this study, antisense strands of siRNA were conjugated to Gal-PEG and mannose 6-phosphate poly(ethylene glycol) (M6P-PEG) for targeted delivery of siRNAs to hepatocytes and hepatic stellate cells (HSCs), respectively. These siRNA conjugates were purified by ion exchange chromatography and verified by gel retardation assay. To evaluate their RNAi functions, the validated siRNA duplexes targeting firefly luciferase and transforming growth factor beta 1 (TGF-β1) mRNA were conjugated to Gal-PEG and M6P-PEG, and their gene silencing efficiencies were determined after transfection into HepG2 and HSC-T6 cells. Disulfide bond between PEG and siRNA was cleaved by dithiothreitol, leading to the release of intact siRNA. Both Gal-PEG-siRNA and M6P-PEG-siRNA conjugates could silence luciferase gene expression for about 40% without any transfection reagents, while the gene silencing effects reached more than 98% with the help of cationic liposomes at the same dose. Conjugation of TGF-β1 siRNA with Gal-PEG and M6P-PEG could silence endogenous TGF-β1 gene expression as well. In conclusion, these siRNA conjugates have the potential for targeted delivery of siRNAs to hepatocytes and hepatic stellate cells for efficient gene silencing in vivo.
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影响因子：
12.4
作者：
Nishina, Kazutaka;Unno, Toshinori;Yokota, Takanori
通讯作者：
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影响因子：
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作者：
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DOI：
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发表时间：
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期刊：
Nature reviews. Drug discovery
影响因子：
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作者：
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通讯作者：
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影响因子：
10.8
作者：
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通讯作者：
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