Egorova Anna, Petrosyan Mariya, Maretina Marianna, Bazian Elena, Krylova Iuliia, Baranov Vladislav, Kiselev Anton
Department of Genomic Medicine, D.O. Ott Research Institute of Obstetrics, Gynecology and Reproductology, Mendeleevskaya Line 3, 199034 Saint-Petersburg, Russia.
Pharmacology Group, D.O. Ott Research Institute of Obstetrics, Gynecology and Reproductology, Mendeleevskaya Line 3, 199034 Saint-Petersburg, Russia.
Pharmaceutics. 2023 Aug 9;15(8):2108. doi: 10.3390/pharmaceutics15082108.
Anti-angiogenic RNAi-based therapy can be considered as a possible strategy for the treatment of endometriosis (EM), which is the most common gynecological disease. Targeted delivery of siRNA therapeutics is a prerequisite for successful treatment without adverse effects. Here we evaluated the RGD1-R6 peptide carrier as a non-viral vehicle for targeted siRNA delivery to endothelial cells in vitro and endometrial implants in vivo. The physicochemical properties of the siRNA complexes, cellular toxicity, and GFP and VEGFA gene silencing efficiency were studied, and anti-angiogenic effects were proved in cellular and animal models. The modification of siRNA complexes with iRGD ligand resulted in a two-fold increase in gene knockdown efficiency and three-fold decrease in endothelial cells' migration in vitro. Modeling of EM in rats with the autotransplantation of endometrial tissue subcutaneously was carried out. Efficiency of anti-angiogenic EM therapy in vivo by anti-VEGF siRNA/RGD1-R6 complexes was evaluated by the implants' volume measurement, CD34 immunohistochemical staining, and VEGFA gene expression analysis. We observed a two-fold decrease in endometriotic implants growth and a two-fold decrease in VEGFA gene expression in comparison with saline-treated implants. RNAi-mediated therapeutic effects were comparable with Dienogest treatment efficiency in a rat EM model. Taken together, these findings demonstrate the advantages of RGD1-R6 peptide carrier as a delivery system for RNAi-based therapy of EM.
基于抗血管生成RNA干扰的疗法可被视为治疗子宫内膜异位症（EM，最常见的妇科疾病）的一种可能策略。靶向递送小干扰RNA（siRNA）疗法是实现无副作用成功治疗的前提条件。在此，我们评估了RGD1-R6肽载体作为一种非病毒载体，用于在体外将siRNA靶向递送至内皮细胞，并在体内递送至子宫内膜植入物。研究了siRNA复合物的物理化学性质、细胞毒性以及绿色荧光蛋白（GFP）和血管内皮生长因子A（VEGFA）基因沉默效率，并在细胞和动物模型中证实了其抗血管生成作用。用整合素靶向激动剂（iRGD）配体修饰siRNA复合物，导致体外基因敲低效率提高两倍，内皮细胞迁移减少三倍。通过皮下自体移植子宫内膜组织对大鼠进行子宫内膜异位症建模。通过测量植入物体积、CD34免疫组化染色和VEGFA基因表达分析，评估抗VEGF siRNA/RGD1-R6复合物在体内对子宫内膜异位症的抗血管生成治疗效果。与生理盐水处理的植入物相比，我们观察到子宫内膜异位植入物生长减少两倍，VEGFA基因表达减少两倍。在大鼠子宫内膜异位症模型中，RNA干扰介导的治疗效果与地诺孕素治疗效果相当。综上所述，这些发现证明了RGD1-R6肽载体作为基于RNA干扰的子宫内膜异位症治疗递送系统的优势。
