Synergistic Anti-Angiogenic Effects Using Peptide-Based Combinatorial Delivery of siRNAs Targeting VEGFA, VEGFR1, and Endoglin Genes

Abstract

Angiogenesis is a process of new blood vessel formation, which plays a significant role in carcinogenesis and the development of diseases associated with pathological neovascularization. An important role in the regulation of angiogenesis belongs to several key pathways such as VEGF-pathways, TGF-β-pathways, and some others. Introduction of small interfering RNA (siRNA) against genes of pro-angogenic factors is a promising strategy for the therapeutic suppression of angiogenesis. These siRNA molecules need to be specifically delivered into endothelial cells, and non-viral carriers modified with cellular receptor ligands can be proposed as perspective delivery systems for anti-angiogenic therapy purposes. Here we used modular peptide carrier L1, containing a ligand for the CXCR4 receptor, for the delivery of siRNAs targeting expression of VEGFA, VEGFR1 and endoglin genes. Transfection properties of siRNA/L1 polyplexes were studied in CXCR4-positive breast cancer cells MDA-MB-231 and endothelial cells EA.Hy926. We have demonstrated the efficient down-regulation of endothelial cells migration and proliferation by anti-VEGFA, anti-VEGFR1, and anti-endoglin siRNA-induced silencing. It was found that the efficiency of anti-angiogenic treatment can be synergistically improved via the combinatorial delivery of anti-VEGFA and anti-VEGFR1 siRNAs. Thus, this approach can be useful for the development of therapeutic angiogenesis inhibition.

Characterization of iRGD-Ligand Modified Arginine-Histidine-Rich Peptides for Nucleic Acid Therapeutics Delivery to αvβ3 Integrin-Expressing Cancer Cells

“…According to the results in Figure 10 b, VEGFA gene expression was decreased by RGD1-mediated anti-VEGFA siRNA delivery by 28.74% and 41.9%, respectively. Previously, we have shown that similar silencing of VEGFA gene expression resulted in significant anti-angiogenic effects in endothelial cell modeland in rat model of endometriosis [ 44 , 46 ]. Thus, RGD1-polyplexes can be suggested as siRNA-carriers in anti-angiogenic therapeutic RNAi.…”

“…Transfection experiments in EA.hy926 cells were performed in triplicate as previously described [ 46 ]. After incubation in a fully supplemented cell culture medium for 48 h, cells were taken for RNA extraction.…”

Modulating the Crosstalk between the Tumor and the Microenvironment Using SiRNA: A Flexible Strategy for Breast Cancer Treatment

“…In this context, delivery of siRNA specifically into endothelial cells is a promising strategy to impair tumor angiogenesis. To this end, a study by Egorova et al [ 69 , 70 ] showed how siRNA-mediated silencing of vascular epithelial growth factor signaling (VEGF-A, VEGFR1) and endoglin, a co-receptor for TGF-β, impaired endothelial cell migration and proliferation. However, an efficient and specific siRNA delivery system is crucial for the development of this promising approach.…”

“…The VEGF–VEGFR axis [ 138 ] plays central roles in angiogenic processes. In order to reduce angiogenetic progression in breast cancer, Egorova et al [ 70 ] used an L1 peptide-based polyplex for the delivery of siRNAs targeting VEGFA, VEGFR1, and endoglin transcripts in CXCR4-positive breast cancer and endothelial cells lines. Polyplexes are polymeric systems in which nucleic acids such as siRNAs are complexed with a cationic polymer through electrostatic interactions [ 139 , 140 ].…”

Drug Delivery of siRNA Therapeutics

“…They used three types of cationic cholesterol derivatives to investigate an optimal formulation to achieve the best performance in terms of gene-silencing and cellular uptake effects. In Egorova et al, [9] researches on modular peptide carriers for the delivery of siRNAs to therapeutic angiogenesis inhibition were performed. In particular, the transfection properties of siRNA as polyplexes were studied in breast cancer cells and endothelial cells.…”