Novel RGD-lipid conjugate-modified liposomes for enhancing siRNA delivery in human retinal pigment epithelial cells
Novel RGD-lipid conjugate-modified liposomes for enhancing siRNA delivery in human retinal pigment epithelial cells
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DOI：
10.2147/ijn.s24447
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发表时间：
2011-01-01
影响因子：
8
通讯作者：
Chiang, Chiao-Hsi
中科院分区：
文献类型：
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作者：
Chen, Cheng-Wei;Lu, Da-Wen;Chiang, Chiao-Hsi
背景：人视网膜色素上皮细胞是小干扰RNA的潜在靶点，可通过抑制血管生成因子的表达来预防和/或治疗脉络膜新生血管；方法：合成一种新型功能性脂类DSPE-Gly(G)-Asp(D)，与1，2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[马来酰亚胺(聚乙二醇2000)]偶联，制备siRNA化RGDPEG化脂质体，以提高其在视网膜色素上皮细胞中的摄取。结果：制备了不同的脂质体，包括1mol%和5mol%的聚乙二醇化脂质和1mol%和5mol%的RGD-聚乙二醇化的脂质体。Zeta-电位分别为：96%，129.7+/-51~230.7+/-60.7 nm，17.3+/-0.6~32+/-1.3mV。在体外视网膜色素上皮细胞研究中，RGD-PEGylated脂质体具有较高的siRNA传递效率，约为PEGylated脂质体的四倍。不同脂质体的比较表明，1mol%RGD修饰的脂质体具有较低的细胞毒性和较高的siRNA递送效率。抗体阻断实验证实，1mol%RGD-PEG化脂质体的摄取是通过整合素受体介导的视网膜色素上皮细胞内吞作用实现的。结论：RGD-PEG化脂质体可通过整合素受体介导的内吞作用将siRNA导入视网膜色素上皮细胞。
Background: Human retinal pigment epithelial cells are promising target sites for small interfering RNA (siRNA) that might be used for the prevention and/or treatment of choroidal neovascularization by inhibiting the expression of angiogenic factor; for example, by downregulating expression of the vascular endothelial growth factor gene.Methods: A novel functional lipid, DSPE-PEG-RGD, a Arg(R)-Gly(G)-Asp(D) motif peptide conjugated to 1, 2-distearoyl-sn-glycero-3-phosphoethanolamine- N-[maleimide (polyethylene glycol)-2000], was synthesized for the preparation of siRNA-loaded RGD-PEGylated liposomes to enhance uptake of encapsulated siRNA in retinal pigment epithelial cells. Various liposomes, with 1 mol% and 5 mol% PEGylated lipid or 1 mol% and 5 mol% RGD-PEGylated lipid, were fabricated.Results: Characterization of the liposomes, including siRNA entrapment efficiency, average particle size and. zeta-potential, were determined to be as follows: >96%, 129.7 +/- 51 to 230.7 +/- 60.7 nm, and 17.3 +/- 0.6 to 32 +/- 1.3 mV, respectively. For the in vitro retinal pigment epithelial cell studies, the RGD-PEGylated liposomes had high delivery efficiency with siRNA delivery, about a four-fold increase compared with the PEGylated liposomes. Comparison of the various liposomes showed that the 1 mol% RGD-modified liposome had less cytotoxicity and higher siRNA delivery efficiency than the other liposomes. The antibody blocking assay confirmed that uptake of the 1 mol% RGD-PEGylated liposome was via integrin receptor-mediated endocytosis in retinal pigment epithelial cells.Conclusion: The results of this study suggest that RGD-PEGylated liposomes might be useful for siRNA delivery into retinal pigment epithelial cells by integrin receptor-medicated endocytosis.