Egorova Anna A, Maretina Marianna A, Kiselev Anton V
D.O. Ott Research Institute of Obstetrics, Gynecology and Reproductology, Saint-Petersburg, Russia.
Saint-Petersburg State University, Saint-Petersburg, Russia.
Methods Mol Biol. 2019;1974:57-68. doi: 10.1007/978-1-4939-9220-1_5.
Discovery of small interfering RNA as a tool for specific gene inhibition led to the development of new therapeutic strategy for the treatment of cancers. The efficacious delivery of therapeutic siRNAs into the cells is a crucial step in RNA interference (RNAi) application, but it remains challenging. Non-viral vectors can provide specific cellular uptake, stable siRNA complex formation, and intracellular siRNA release. Recently, we evaluated modular peptide carrier L1 bearing CXCR4 targeting ligand for its ability to condense siRNA and facilitate endosomal escape and VEGFA gene silencing in CXCR4-expressing endothelial and glioblastoma cells. The present chapter showcases the ability of L1 targeted peptide carrier to form complexes with siRNA and provide efficient VEGFA gene knockdown. We showed that siRNA delivery by means of L1 peptide carrier can result in significant decrease of VEGFA gene expression in A172 glioblastoma cells and in EA.hy 926 endothelial cells. Also, delivery of anti-VEGFA siRNA/peptide complexes led to significant inhibition of endothelial cell migration. Our results showed that L1 peptide carrier modified with CXCR4 ligand is a promising tool for targeted siRNA delivery into CXCR4-expressing cancer and endothelial cells.
小干扰RNA作为一种特异性基因抑制工具的发现，促使了癌症治疗新策略的发展。将治疗性小干扰RNA有效递送至细胞内是RNA干扰（RNAi）应用中的关键步骤，但这仍然具有挑战性。非病毒载体能够实现特异性细胞摄取、稳定的小干扰RNA复合物形成以及细胞内小干扰RNA释放。最近，我们评估了携带CXCR4靶向配体的模块化肽载体L1在CXCR4表达阳性的内皮细胞和成胶质细胞瘤细胞中浓缩小干扰RNA、促进内体逃逸以及沉默VEGFA基因的能力。本章展示了L1靶向肽载体与小干扰RNA形成复合物并有效敲低VEGFA基因的能力。我们发现，通过L1肽载体递送小干扰RNA可导致A172成胶质细胞瘤细胞和EA.hy 926内皮细胞中VEGFA基因表达显著降低。此外，抗VEGFA小干扰RNA/肽复合物的递送导致内皮细胞迁移受到显著抑制。我们的结果表明，用CXCR4配体修饰的L1肽载体是一种将小干扰RNA靶向递送至CXCR4表达阳性的癌细胞和内皮细胞的有前景的工具。
