Glycopolymers/PEI complexes as serum-tolerant vectors for enhanced gene delivery to hepatocytes
Glycopolymers/PEI complexes as serum-tolerant vectors for enhanced gene delivery to hepatocytes
复制标题
糖聚合物/PEI 复合物作为血清耐受载体，用于增强向肝细胞的基因递送
DOI：
10.1016/j.carbpol.2018.10.036
复制
发表时间：
2019-02-01
影响因子：
11.2
通讯作者：
Shi, Tongfei
中科院分区：
文献类型：
--
作者：
Zhao, Liman;Li, Yanchun;Shi, Tongfei
血清稳定性是理想的聚合基因载体的关键因素。本工作设计了一系列耐血清、低毒性的糖共聚物/聚乙烯亚胺（PEI）复合物用于基因传递。采用原子转移自由基聚合法（ATRP）合成了梳形无规共聚物葡聚糖-聚（2-甲基丙烯酸二甲氨基乙酯-co-2-甲基丙烯酸乳糖酰胺乙酯）（DDrL）。然后研究了DDrLs/PEI作为质粒DNA（pDNA）载体的用途，其可以将pDNA完全浓缩成纳米颗粒。DDrL/PEI/pDNA复合物在含血清培养基中显示出比PEI/ pDNA复合物更好的稳定性。体外基因转染研究表明，与无血清条件下相比，在血清存在下，DDrL/PEI表现出显著的转染效率增强。此外，在30%血清存在下，DDrLs/PEI的转染水平比单独PEI高两个数量级。DDrLs/PEI与半乳糖的复合物增强了pDNA向肝细胞的递送，在ASGPr呈递的HepG 2中的蛋白表达高于缺乏受体的HeLa细胞。所有DDrL/PEI/pDNA复合物的细胞毒性均低于PEI/pDNA。
Serum stability is a crucial factor for ideal polymeric gene vectors. In this work, a series of serum-tolerant and low-toxicity glycopolymers/poly(ethyleneimine) (PEI) complexes were designed for gene delivery. Atomic transfer radical polymerization (ATRP) was used to synthesize the comb-shaped random copolymers dextran-gpoly(2-dimethylaminoethyl methacrylate-co-2-lactobionamidoethyl methacrylate) (DDrL). Then DDrLs/PEI were investigated for their use as plasmid DNA (pDNA) vectors, which can completely condense the pDNA into nanoparticles. The DDrLs/PEI/pDNA complexes in serum-containing media showed better stability than PEI/ pDNA complexes. in vitro gene transfection studies showed that DDrLs/PEI exhibited a remarkable transfection efficiency enhancement in the presence of serum compared to that in serum-free conditions. Moreover, the transfection level of DDrLs/PEI were two orders of magnitude higher than that of PEI alone in the presence of 30% serum. DDrLs/PEI complexes with galactose enhanced pDNA delivery to hepatocytes, with higher protein expression in ASGPr-presenting HepG2 than in HeLa cells, which lack the receptor. All of the DDrLs/PEI/pDNA complexes had lower cytotoxicity than PEI/pDNA.