Combination of TAT-HMGB1A and R3V6 amphiphilic peptide for plasmid DNA delivery with anti-inflammatory effect

Abstract

High mobility group box 1 (HMGB1) is not only a non-histone nuclear protein but also a pro-inflammatory cytokine. Previously, it has been shown that HMGB1 box A domain (HMGB1A) is an antagonist of HMGB1. HMGB1A can deliver plasmid DNA (pDNA) into cells due to its positive charge and ability to form complexes with pDNA. In this study, TAT-linked HMGB1A (TAT-HMGB1A) was produced using recombinant DNA technology. The pDNA/TAT-HMGB1A/R3V6 ternary complex was then prepared for efficient delivery of pDNA by coating the pDNA/HMGB1A complex with the R3V6 amphiphilic peptide. The particle size of the pDNA/TAT-HMGB1A/R3V6 complex was approximately 120nm and had the highest delivery efficiency at a 1: 5: 15 weight ratio. The pDNA/TAT-HMGB1A/R3V6 complex had a higher transfection efficiency than the pDNA/poly-L-lysine (PLL), pDNA/R3V6, and pDNA/TAT-HMGB1A complexes. In the RAW 264.7 cells activated by lipopolysaccharides, the TNF-alpha and IL-6 levels were decreased by the addition of the pDNA/TAT-HMGB1A/R3V6 complex, suggesting that TAT-HMGB1A maintained the anti-inflammatory effect of HMGB1A. Delivery of the heme oxygenease-1 (HO-1) gene using TAT-HMGB1A/R3V6 further decreased the pro-inflammatory cytokines, due to the anti-inflammatory effect of HO-1. The results suggest that the pDNA/ TAT-HMGB1A/R3V6 complex may be a useful gene carrier for gene therapy of inflammatory diseases.