An efficient PEGylated gene delivery system with improved targeting: Synergism between octaarginine and a fusogenic peptide

Abstract

Because of their ability to translocate different cargos into cells, arginine-rich cell penetrating peptides (CPPs) are promising vehicles for drug and gene delivery. The use of CPP-based carriers, however, is hampered by the lack of specificity and by interactions with negative serum components. Polyethylene glycol (PEG) is used to decrease such non-specific interactions, albeit its use is associated with reduced transfection efficiency. In this study, we describe the development of PEGylated CPP-based gene carrier with an improved targeting and a high transfection activity. The system was prepared by condensing DNA with a polycation followed by coating with a lipid envelope containing the octaarginine (R8) peptide as a model CPP. R8-modified nanoparticles produced high transfection activities, but the efficiency was reduced by PEG shielding. The reduced activity could be fully restored by the addition of a targeting ligand and a pH-sensitive fusogenic peptide. The efficiency of the proposed system is quite high, even in the presence of serum, and shows improved targeting and selectivity. Surprisingly, the effect of the fusogenic peptide was dramatically reduced in the absence of R8. Although shielded, R8 augmented the activity of the fusogenic peptide, suggesting a synergistic effect between the two peptides at the intracellular level.

Lipid Nanoparticles for Cell-Specific <i>in Vivo</i> Targeted Delivery of Nucleic Acids

“…One example of LPHNPs is the multifunctional envelope-type nanodevice (MEND) first described by Kogure et al 42) The MEND system was upgraded with various modifications to enhance the efficiency of gene delivery in vitro and in vivo. [43][44][45] Remarkably, the use of octaatgrinine (R8) peptide significantly enhanced the cellular uptake and nuclear delivery, and the use of GALA peptide enhanced endosomal escape. [46][47][48] The combination of R8 and GALA peptides produced very efficient MENDs based on synergism between the two peptides.…”

“…[43][44][45] Remarkably, the use of octaatgrinine (R8) peptide significantly enhanced the cellular uptake and nuclear delivery, and the use of GALA peptide enhanced endosomal escape. [46][47][48] The combination of R8 and GALA peptides produced very efficient MENDs based on synergism between the two peptides. 49) Different MENDs were prepared encapsulating pDNA, siRNA, or mRNA and could be successfully used for efficient gene delivery in vitro and in vivo.…”

Nucleic acid delivery to differentiated retinal pigment epithelial cells using cell-penetrating peptide as a carrier

“…Cell penetrating peptides are one potential class of non-viral gene delivery systems; these can be either cationic or amphipathic in nature. These peptides have cell penetrating segments and they have been used to deliver small molecules, antibodies and nucleic acids into cells [6][7][8][9].…”

DNA Vaccines—How Far From Clinical Use?

“…However, most of these studies have been performed under serum-poor conditions in vitro, and the possibility of rapid formation of a protein corona around the NC and engagement of negatively charged serum components by CPP prior to cellular engagement needs to be taken into account [ 134 ]. NC surface modifications like dense decoration with polyethylene glycol (PEG) have been introduced to limit serum protein adsorption, and to retain CPP activity [ 135 ]. Major safety concerns associated with the use of NC are are biocompatibility, biodistributiion and clearance, and the induction of unwanted immune reactions [ 136 ].…”