RGD-POLYMER TARGETING PLASMID TO ANGIOGENIC ENDOTHELIUM

描述(由申请人提供)。项目摘要在目前的应用中，合成了PEI-PEG-RGD靶向基因载体并进行了表征。将构建的编码sFllt-1的表达载体携带到新生血管真皮微血管内皮细胞中。体内研究表明，该递送系统携带的sFlt-1表达延缓了肿瘤的生长，提高了动物的存活率。因此，PEI-PEG-RGD/pCMV-sFlt-1复合体可用于肿瘤特异性抗血管生成基因治疗。此外，还构建了IL-2与sFlk-1的融合表达载体。体外和体内实验表明，该化合物具有良好的抗肿瘤活性。在这一新的应用中，我们建议传递siRNA来沉默血管内皮生长因子及其受体。RGD将偶联到可还原的聚合物聚(CBA-DAH)上，这种聚合物是已知的无毒的，可以极大地提高几种细胞系的转染率。RGD-CHOL-R9C也将被合成，因为CHOL-R9C已经证明了siRNA的高转染率。此外，还将设计RGD-聚乙二醇水溶脂聚合物(WSLP)，因为WSLP已被本实验室证明是一种良好的载体。这些与siRNA络合的RGD偶联聚合物将进行广泛的体外和体内动物研究。在小鼠前列腺癌和乳腺癌模型中，肿瘤生长的抑制将在结果中得到证实。所获得的数据可用于设计治疗癌症患者的siRNA载体。与公共健康相关：三种新设计的聚合物将与RGD结合。预计体内和体外的研究都将为siRNA传递带来积极的结果，并可应用于肿瘤治疗。

DESCRIPTION (provided by applicant). PROJECT SUMMARY ABSTRACT In the current application, PEI-PEG-RGD targeted gene carrier was synthesized and characterized. The constructed plasmid encoding sFllt-1 was carried to angiogenic dermal microvascular endothelial cells. The in vivo studies demonstrated that expressed sFlt-1 carried by this delivery system delayed tumor growth and increased survival rate of animal. Therefore, PEI-PEG-RGD/pCMV- sFlt-1 complex can be useful to develop tumor specific anti-angiogenic gene therapy. In addition, combination of IL-2 and sFlk-1 plasmid was constructed. The in vitro and in vivo studies demonstrated excellent efficiency of anti-tumor activity. In this renewal application, we propose to deliver siRNA for silencing VEGF and VEGF receptors. RGD will be conjugated to the reducible polymer, poly (CBA-DAH), which is known to be non-toxic and greatly enhances transfection in several cell lines. RGD-chol-R9C will also be synthesized, since chol-R9C has demonstrated high transfection of siRNA. In addition, RGD- PEG-water soluble lipopolymer (WSLP) will be designed as WSLP has been proven an excellent carrier in this laboratory. Extensive in vitro and in vivo animal studies will be carried out with these RGD-conjugated polymers complexed with siRNA. Inhibition of tumor growth in murine prostate and breast adenocarcinoma models will be proven in the results. The obtained data can be utilized for the design of siRNA delivery to treat cancer patients. PUBLIC HEALTH RELEVANCE: Three newly designed polymers will be conjugated with RGD. It is expected that both in vitro and in vivo studies will present positive results for siRNA delivery, which can be applied to tumor treatment.