Efficient siRNA Delivery Using PEG-conjugated PAMAM Dendrimers Targeting Vascular Endothelial Growth Factor in a CoCl2-induced Neovascularization Model in Retinal Endothelial Cells.
Efficient siRNA Delivery Using PEG-conjugated PAMAM Dendrimers Targeting Vascular Endothelial Growth Factor in a CoCl2-induced Neovascularization Model in Retinal Endothelial Cells.
复制标题
在 CoCl2 诱导的视网膜内皮细胞新血管形成模型中，使用 PEG 缀合的 PAMAM 树状聚合物靶向血管内皮生长因子进行高效 siRNA 递送。
DOI：
10.2174/1567201812666150817123049
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发表时间：
2016-05
期刊：
影响因子：
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通讯作者：
Rong Qi
中科院分区：
文献类型：
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作者：
Xinlei Wang;George Liu;Yong Tao;Rong Qi
背景
脉络膜新生血管(CNV)，也被称为视网膜下新生血管，由于更常见于黄斑，会对中心视力造成严重损害。血管生成中最重要的因子是血管内皮生长因子(VEGF)。通过RNAi技术，可将血管内皮生长因子基因敲除用于治疗新生血管。聚乙二醇偶联胺(PEGPAMAM)树枝状大分子作为一种新型的合成聚合物，在基因传递载体方面具有广阔的应用前景。
方法
为了研究聚乙二醇胺树状大分子的siRNA传递效率，我们制备了带有荧光标记siRNA的聚乙二醇胺树状大分子树枝状网络(PEG-PamAM/FAM siRNA)或血管内皮生长因子siRNA(PEG-PamAM/VEGFsiRNA)，并研究了树突状网络在氯化钴(CoCl2)诱导的视网膜血管内皮细胞(RF/6A)新生血管模型中的转染和下调效应。
结果
我们的结果表明，聚乙二醇-PAMAM树状大分子对FAM siRNA的转染率显著高于商业转染剂PEI1.4倍(P<；0.001)。与PEI/VEGFsiRNA复合体相比，PEI-PamAm/VEGFsiRNA树枝状复合体在mRNA和蛋白表达水平上均显著下调了VEGFmRNA的表达(P<；0.01)。管形成实验还证明，在RF/6A中，聚乙二醇-PAMAM/VEGFsiRNA树状网络比PEI/VEGFsiRNA更显著地抑制血管样形成(P<；0.001)。
结论
本研究表明，G5-PEG在促进siRNA转运、下调血管内皮生长因子的表达和抑制RF/6A上的血管样形成方面比PEI更有效。
BACKGROUND
Choroidal neovascularization (CNV), also known as subretinal neovascularization, causes serious damage to the central vision as it happens more commonly in macula. The most important factor involved in angiogenesis is vascular endothelial growth factor (VEGF). By an RNAi technique, VEGF gene knockdown can be used to treat CNV. PEG-conjugated poly (amidoamine) (PEGPAMAM) dendrimers as a new type of synthetic polymers are very promising to be gene delivery carriers.
METHODS
To investigate siRNA delivery efficacy of PEG-PAMAM dendrimers, we prepared dendriplexes of PEG-PAMAM dendrimers with a fluorescence-labelled siRNA (PEG-PAMAM/FAM siRNA) or VEGF siRNA (PEG-PAMAM/VEGF siRNA), and studied transfection and downregulation efficacy of the dendriplexes in a cobalt chloride (CoCl2)-induced neovascularization model in retinal vascular endothelial cells (RF/6A).
RESULTS
Our results demonstrate that PEG-PAMAM dendrimers had significantly higher transfection efficiency to FAM siRNA than a commercial transfection reagent PEI (1.4-fold, P<0.001) measured by flow cytometry. Compared to the PEI/VEGF siRNA polyplexes, the dendriplexes of the PEG-PAMAM/VEGF siRNA more significantly downregulated VEGF gene expression (P < 0.01) at both mRNA and protein expression level. A tube formation assay also proved that the PEG-PAMAM/VEGF siRNA dendriplexes more significantly inhibited vascular-like formation than PEI/VEGF siRNA did (P < 0.001) in RF/6A.
CONCLUSION
This study demonstrated that G5-PEG was more efficient than PEI in facilitating siRNA delivery, downregulating VEGF expression and inhibiting vascular-like formation on RF/6A.