肿瘤靶向型联合基因治疗系统的制备及生物医学应用

项目名称： 肿瘤靶向型联合基因治疗系统的制备及生物医学应用

项目编号： No.51303137

项目类型： 青年科学基金项目

立项/批准年度： 2014

项目学科： 一般工业技术

项目作者： 曾旋

作者单位： 武汉大学

项目金额： 25万元

中文摘要： 制备两类不同系列的具有肿瘤靶向功能、抗肿瘤功效以及良好生物安全性的非病毒联合基因治疗系统。体系1：以亲和素为基体，通过生物素-亲和素的特异性作用，将固相合成的生物素化细胞穿膜肽偶联到亲和素表面，并与两种治疗基因通过静电作用组装得到复合物，最后在复合物表面引入生物素化人转铁蛋白作为肿瘤靶向基团，得到具有肿瘤靶向功能的双基因联合治疗系统；体系2：将生物素化穿膜肽/治疗基因复合物、肿瘤坏死因子相关凋亡诱导配体(TRAIL)以及人转铁蛋白同时偶联到亲和素表面，得到肿瘤靶向型治疗基因/TRAIL联合治疗系统。选用p53抑癌基因和抑制肿瘤血管生成的小分子干扰RNA作为治疗基因。研究这两类系统的体外细胞毒性、转染效率、肿瘤特异性以及杀瘤能力。优化穿膜肽、治疗基因、凋亡因子和靶向基团的用量比例，筛选出效果显著的体系进行动物实验。建立荷人癌细胞移植瘤的裸鼠模型，研究各系统在动物体内的毒性反应及抗肿瘤效果。

中文关键词： 肿瘤靶向；联合治疗；基因治疗；多肽；刺激响应性

英文摘要： Two series of nonviral combined gene therapy systems, presented tumor targeting ability, anti-tumor activity and bio-safety, will be prepared in this study. System 1: Avidin will be used as matrix, and various biotinylated cell-penetrating peptides (CPPs) obtained from solid-phase synthesis step will be coupled to avidin surface, which attributed to the avidin-biotin bridge. Then, two therapeutic genes with different anti-tumor activities will be complexed with CPP-avidin bioconjugates via electrostatic interaction. After that, biotinylated transferrin used as tumor targeted ligand will cover the complexes, and finally a series of tumor targeted combined gene therapy systems will be formed. System 2: By using avidin-biotin bridge, various biotinylated CPPs/therapeutic gene complexes, biotinylated TRAIL and biotinylated transferrin will be coupled to avidin surface, to obtain a series of tumor targeted therapeutic gene/TRAIL combined therapy systems. Plasmid pDsRed2-N1-p53 and VEGF siRNA will be used as therapeutic genes. Characteristics of these obtained biotinylated CPPs and combined gene therapy systems will be evaluated in terms of agarose gel electrophoresis, SEM, particle size and ζ-potential measurements. Their in vitro cell viability and transfection ability will be investigated in different cell lines. A

英文关键词： tumor-targeting；combined therapy；gene therapy；peptide；stimuli-responsive