Degradable polyethylenimine derivate coupled to a bifunctional peptide R13 as a new gene-delivery vector
Degradable polyethylenimine derivate coupled to a bifunctional peptide R13 as a new gene-delivery vector
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DOI：
10.2147/ijn.s28819
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发表时间：
2012-01-01
影响因子：
8
通讯作者：
Gao, Shen
中科院分区：
文献类型：
--
作者：
Liu, Kehai;Wang, Xiaoyu;Gao, Shen
背景：为了解决聚乙烯亚胺(PEI)作为非病毒基因载体的细胞毒性和肿瘤靶向性问题，开发了一种可降解的PEI衍生物，将其偶联到双功能肽R13上。方法：首先，将小分子PEI与Pluronic P123交联，合成可降解的PEI衍生物，然后将肿瘤靶向多肽精氨酸-甘氨酸-天冬氨酸-半胱氨酸(RGDC)与细胞穿透性多肽TAT(49-57)结合，得到具有促进细胞选择和细胞摄取的双功能多肽RGDC-TAT(49-57)采用R13对PEI衍生物进行修饰，制备了新型聚合基因载体(P123-PEI-R13)。对新基因载体的化学结构和生物物理参数进行了表征。结果：与PEI 25 KDa相比，该载体对Hela细胞和小鼠黑色素瘤B16细胞的降解受控，靶向性强，对Hela细胞和B16细胞无细胞毒性。P123-PEI-R13/DNA络合物的粒径约为100-250 nm，具有合适的Zeta电位。当DNase I/mgDNA的浓度为6U时，该纳米粒子可以保护DNA不被DNase I消化。纳米粒对50%胎牛血清和600 mU g/mL肝素钠诱导的解离有较好的耐受性。P123-PEI-R13在两种细胞系中的转染率均高于PEI 25KDA。结论：P123-PEI-R13有望成为一种安全有效的基因治疗载体。
Background: To solve the efficiency versus cytotoxicity and tumor-targeting problems of polyethylenimine (PEI) used as a nonviral gene delivery vector, a degradable PEI derivate coupled to a bifunctional peptide R13 was developed.Methods: First, we synthesized a degradable PEI derivate by crosslinking low-molecular-weight PEI with pluronic P123, then used tumor-targeting peptide arginine-glycine-aspartate-cysteine (RGDC), in conjunction with the cell-penetrating peptide Tat (49-57), to yield a bifunctional peptide RGDC-Tat (49-57) named R13, which can improve cell selection and increase cellular uptake, and, lastly, adopted R13 to modify the PEI derivates so as to prepare a new polymeric gene vector (P123-PEI-R13). The new gene vector was characterized in terms of its chemical structure and biophysical parameters. We also investigated the specificity, cytotoxicity, and gene transfection efficiency of this vector in alpha v beta 3-positive human cervical carcinoma Hela cells and murine melanoma B16 cells in vitro.Results: The vector showed controlled degradation, strong targeting specificity to alpha v beta 3 receptor, and noncytotoxicity in Hela cells and B16 cells at higher doses, in contrast to PEI 25 KDa. The particle size of P123-PEI-R13/DNA complexes was around 100-250 nm, with proper zeta potential. The nanoparticles can protect plasmid DNA from being digested by DNase I at a concentration of 6 U DNase I/mu g DNA. The nanoparticles were resistant to dissociation induced by 50% fetal bovine serum and 600 mu g/mL sodium heparin. P123-PEI-R13 also revealed higher transfection efficiency in two cell lines as compared with PEI 25 KDa.Conclusion: P123-PEI-R13 is a potential candidate as a safe and efficient gene-delivery carrier for gene therapy.