Self‐Assembly of a Multifunctional Lipid With Core–Shell Dendrimer DNA Nanoparticles Enhanced Efficient Gene Delivery at Low Charge Ratios into RPE Cells

Abstract

Gene therapy is a promising approach to treat genetic diseases. Development of gene delivery systems with high efficiency and low toxicity is a high priority in clinical application of gene therapy. In this work, we designed and evaluated a hybrid nonviral gene delivery system composed of a multifunctional lipid ECO and a core-shell dendrimer G4 nanoglobule for efficient intracellular gene delivery at low charge ratios (N/P). The dendrimeric nanoglobule was designed to effectively condense the gene cargo and the cationic lipid was used to enhance cellular uptake. As expected, this hybrid system formed stable nanoparticles at low N/P ratios (overall N/P = 6) and had low cytotoxicity. The hybrid nanoparticles induced higher GFP (green fluorescent protein) expression in ARPE-19 cells in the culture medium containing 10% serum than in serum-free medium at low N/P ratios. The nanoparticles also mediated significant reporter gene GFP expression in the retina ex-vivo from wild type C57 mice and in vivo with BALB/c mice. Together, these results demonstrate that the rationally designed G4 nanoglobule/pDNA/ECO nanoparticles are a promising delivery system for in vitro and in vivo gene delivery at low charge ratios.

Targeted Multifunctional Lipid ECO Plasmid DNA Nanoparticles as Efficient Non-viral Gene Therapy for Leber’s Congenital Amaurosis

“…ECO is a multifunctional lipid that has demonstrated excellent efficiency for cytosolic delivery of a variety of genetic materials because of its intrinsic PERC effect 22, 23, 24, 25, 26. This study has shown that ECO is also effective for delivering therapeutic pDNA for non-viral gene replacement therapy in Rpe65 −/− mice.…”

“…This study has shown that ECO is also effective for delivering therapeutic pDNA for non-viral gene replacement therapy in Rpe65 −/− mice. The superior transfection properties of ECO/pDNA nanoparticles result from the multifunctional properties of the lipid carrier, including self-assembly formation of stable nanoparticles with pDNA without helper lipids, pH-sensitive amphiphilic cell membrane destabilization and endosomal escape, as well as reductive dissociation of the nanoparticles to release nucleic acids in the cytoplasm 22, 26. Here we tested a targeting mechanism that involves the use of IRBP to enhance pDNA delivery with ECO into RPE cells (Figure 8).…”

“…To assess GFP expression, the mice were sacrificed, and eyes were collected 3 days after injection, washed with penicillin-streptomycin solution (Sigma), and rinsed with Hank’s balanced salt solution (HyClone). Eye cups were prepared as described previously 26 . The retina and RPE layers were placed in glass-bottom confocal plates and fixed with 1 mL of PBS containing 4% paraformaldehyde.…”

“…Recently, we designed a multifunctional lipid, (1-aminoethyl)iminobis[N-(oleicylcysteinyl-1-amino-ethyl)propionamide] (ECO), as a simple and smart gene delivery carrier based on its mechanism of pH-sensitive amphiphilic endosomal escape and reductive cytosolic release (PERC) of nucleic acids 22, 23, 24, 25, 26. ECO contains a protonable ethylenediamine (E) head group, two cysteine (C) functional linkers, and two oleoyl (O) lipophilic tails.…”

Role of eIF4E on epithelial‐mesenchymal transition, invasion, and chemoresistance of prostate cancer cells

“…ECO/siRNA nanoparticles have been shown to mediate effective RNAi of various oncogenic targets in vitro and in vivo [8‐10]. To further confirm the role of eIF4E in PCa cells, the effects of down‐regulation of eIF4E were assessed using RNAi based on ECO/siRNA nanoparticles.…”