Coupling of a bifunctional peptide R13 to OTMCS-PEI copolymer as a gene vector increases transfection efficiency and tumor targeting.
Coupling of a bifunctional peptide R13 to OTMCS-PEI copolymer as a gene vector increases transfection efficiency and tumor targeting.
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DOI：
10.2147/ijn.s59726
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发表时间：
2014
影响因子：
8
通讯作者：
Liu K
中科院分区：
文献类型：
--
作者：
Lv H;Zhu Q;Liu K;Zhu M;Zhao W;Mao Y;Liu K
开发了一种与双功能肽R13偶联的可降解聚乙烯亚胺（PEI）衍生物，以解决PEI用作基因载体时的转染效率与细胞毒性和肿瘤靶向问题。将低分子量聚乙烯亚胺（PEI）与N-辛基-N-季铵化壳聚糖（OTMCS）交联，合成可降解聚乙烯亚胺衍生物（OTMCS-PEI），再用双功能肽RGDC-TAT（49-57）R13修饰OTMCS-PEI，制备新型基因载体OTMCS-PEI-R13。通过各种理化方法对这种新的基因载体进行了表征。并测定了其细胞毒性和基因转染效率。该载体表现出可控的降解和优异的缓冲能力。OTMCS-PEI-R13/DNA复合物的粒径为约150-250 nm，ζ电位范围为10 mV至30 mV。在23.5UDNase I/μ gDNA的浓度下，该聚合物可保护质粒DNA不被DNase I消化。此外，聚合物对50%胎牛血清和400 μg/mL肝素钠诱导的解离具有抗性。与PEI 25 kDa相比，OTMCS-PEI-R13/DNA复合物在体外和体内均表现出更高的转染效率。此外，与OTMCS-PEI相比，OTMCS-PEI-R13在肿瘤部位的分布显著增强，表明R13的肿瘤靶向特异性。OTMCS-PEI-R13有望成为一种安全、高效的基因治疗载体。
A degradable polyethylenimine (PEI) derivative coupled to a bifunctional peptide R13 was developed to solve the transfection efficiency versus cytotoxicity and tumor-targeting problems of PEI when used as a gene vector. We crossed-linked low molecular weight PEI with N-octyl-N-quaternary chitosan (OTMCS) to synthesize a degradable PEI derivative (OTMCS-PEI), and then used a bifunctional peptide, RGDC-TAT (49–57) called R13 to modify OTMCS-PEI so as to prepare a new gene vector, OTMCS-PEI-R13. This new gene vector was characterized by various physicochemical methods. Its cytotoxicity and gene transfection efficiency were also determined both in vitro and in vivo. The vector showed controlled degradation and excellent buffering capacity. The particle size of the OTMCS-PEI-R13/DNA complexes was around 150–250 nm and the zeta potential ranged from 10 mV to 30 mV. The polymer could protect plasmid DNA from being digested by DNase I at a concentration of 23.5 U DNase I/μg DNA. Further, the polymer was resistant to dissociation induced by 50% fetal bovine serum and 400 μg/mL sodium heparin. Compared with PEI 25 kDa, the OTMCS-PEI-R13/DNA complexes showed higher transfection efficiency both in vitro and in vivo. Further, compared with OTMCS-PEI, distribution of OTMCS-PEI-R13 at tumor sites was markedly enhanced, indicating the tumor-targeting specificity of R13. OTMCS-PEI-R13 could be a potential candidate as a safe and efficient gene delivery carrier for gene therapy.