Efficient downregulation of VEGF in retinal pigment epithelial cells by integrin ligand-labeled liposome-mediated siRNA delivery

Abstract

Background:The purpose of this study was to demonstrate the effectiveness of an integrin peptide ligand-labeled liposomal delivery system loaded with vascular endothelial growth factor (VEGF)-siRNA in a model study of gene therapy for retinopathy using human retinal pigment epithelial cells. Methods: Arg(R)-Gly(G)-Asp(D) motif peptide conjugating polyethylene glycol modified (RGD-PEGylated) liposomes were prepared using a thin-film hydration method and optimized for surface charge, particle size, small interfering RNA (siRNA) load, and entrapment efficiency. Reverse transcriptase-polymerase chain reaction and enzyme-linked immunosorbent assays were used to determine VEGF levels in retinal pigment epithelial cells. Cytotoxicity was determined using the 3-[4, 5-dimethylthiazol-2-yl]-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay and flow cytometry. Results: Physicochemical properties, including particle size, zeta potential, and siRNA load, of the prepared RGD-PEGylated liposomes and their entrapment efficiency were determined to be within the following ranges: 123.8-234.1 nm, 17.31-40.09 mV, 5.27%-6.33%, and .97%, respectively. RGD-PEGylated liposome-mediated fluorescent-labeled siRNA delivery demonstrated significantly enhanced cellular uptake, and 3 mol% RGD-PEGylated liposomes (having 3β-[N-(N', N'-dimethylaminoethane) carbamoyl] cholesterol (DC-cholesterol) DSPE and DSPE-PEG(2000)-RGD with molar ratio of 50/47/3) were shown to have better efficacy with regard to specificity for retinal pigment epithelial cells, reduced cytotoxicity, and knockdown of the target molecule. Conclusion: By integrin receptor-mediated endocytosis, 3 mol% RGD-PEGylated liposomes were shown to be a suitable vector when loaded with VEGF-siRNA for efficient downregulation of VEGF in retinal pigment epithelial cells at both the protein and gene levels. This integrin ligand-modified liposomal delivery system has therapeutic potential for ocular gene therapy.

<p>Codelivery of GRP78 siRNA and docetaxel via RGD-PEG-DSPE/DOPA/CaP nanoparticles for the treatment of castration-resistant prostate cancer</p>

“…CaP-based nanoparticles are especially widely used for siRNA delivery, due to their good biocompatibility and high delivery efficiency. Since neovascular vessels are abundant in tumors, and αvβ3 integrin is highly expressed in cancer, we used the RGD peptide as a target moiety in our study 23,38. In this study, we prepared a novel LCP-RGD targeted nanoparticle, which can effectively encapsulate and then slowly release the siRNA and DTXL.…”

“…However, due to its poor in vivo stability and poor cellular uptake, this approach requires an effective gene delivery vehicle to deliver the siRNA to target cancer cells 19. Over the past few years, various nanoparticle delivery vectors have been designed to effectively deliver chemotherapeutic agent and siRNA 20–23. Among them, calcium phosphate (CaP) nanoparticles have attracted much attention with regard to its use in siRNA delivery 24,25.…”

Preparation and optimisation of anionic liposomes for delivery of small peptides and cDNA to human corneal epithelial cells

“…Using a 4:2:1 ratio of PC/CAP/DOPE, we have developed an anionic liposome preparation with a hydrodynamic size within the range for optimal peptide delivery (Chen et al, 2013, Allon et al, 2012). While larger particles can encapsulate more cargo, liposome uptake is highly dependent on and inversely correlated with particle size.…”

Nanotechnology for Age-Related Macular Degeneration

“…harmaceutics 2021, 13, x FOR PEER REVIEW continuously, so as to broadly solve non-hereditary retinal diseases, such as AMD. et al proved that RGD-PEGylated liposomes loaded with VEGF-siRNA can effec reduce the level of VEGF in ARPE-19 cells and have therapeutic potential for ophth gene therapy [46]. Takashima et al prepared pDNA/PEI composite liposomes, w showed high nucleic acid encapsulation efficiency and cell absorption capacity in A 19 cells, which significantly improved the efficacy of posterior gene therapy [47] worth noting that liposomes injected intravenously into the eye will not dist smoothly and remain in the extensive lymphatic vascular network [48].…”