Stability of Dendriplexes Formed by Anti-HIV Genetic Material and Poly(propylene imine) Dendrimers in the Presence of Glucosaminoglycans

Abstract

There are several barriers to the application of dendriplexes formed by poly(propylene imine) dendrimers and genetic material for gene therapy. One limitation is their interaction with extracellular matrix components such as glucosaminoglycans. These can displace the genetic material from the dendriplexes, affecting their transfection activity. In this study, we analyzed the interaction between dendriplexes and the four main glucosaminoglycans (heparin, heparan sulfate, chondroitin sulfate, and hyaluronic acid) by fluorescence polarization and gel electrophoresis. Dendriplexes were formed by combining three anti-HIV antisense oligodeoxynucleotides with three poly(propylene imine) dendrimers of the fourth generation: unmodified and partially modified with maltose and maltotriose (open shell glycodendrimers). The data showed that the effect of glucosaminoglycans on dendriplexes depends on the glucosaminoglycan type and the oligosaccharide serving as the surface group of the dendrimer. Heparin at physiological concentrations destroys dendriplexes formed by open shell glycodendrimers, but dendriplexes based on unmodified poly(propylene imine) dendrimers are stable in its presence. The other glucosaminoglycans at physiological concentrations cannot destroy dendriplexes formed by any of the dendrimers studied.

Dendritic glycopolymers based on dendritic polyamine scaffolds: view on their synthetic approaches, characteristics and potential for biomedical applications

“…Due to their negative charge, they can displace oligonucleotide cargo from dendriplexes, thus seriously affecting the transfection process. For this reason, Szewczyk et al 265 between complexes formed by PPI G3 (naked, maltose-and maltotriose-modified) and anti-HIV ODNs (AT, SREV and GEM91) and four GAGs (heparin, heparan sulfate, chondroitin sulfate, and hyaluronic acid). Addition of heparin, possessing the highest negative charge of all GAGs, to unmodified PPI-ODNs complexes had no or only a slight effect up to 10 mg mL À1 as assessed by the fluorescence polarization method.…”

“…On the other hand, while naked PPI-ODNs complexes are unaffected by physiological concentrations of heparin in the absence of cell medium, glycodendrimer-ODNs complexes undergo dissociation in the presence of these GAGs. 265…”

The AGMA1 poly(amidoamine) inhibits the infectivity of herpes simplex virus in cell lines, in human cervicovaginal histocultures, and in vaginally infected mice

“…Polycationic dendrimers have been so far developed mainly for the transfection of genetic material into eukaryotic cells for gene therapy, an approach that has been found however to be burden by the tendency of these dendrimers to bind to glycosaminoglycans of the cells surface [31]. With these premises, some HSPG-targeting polycationic dendrimers have been developed and assayed in vitro against different viruses, showing promising features: the peptide dendrimer SB105-A10, containing clusters of basic amino acids, proved to be a potent inhibitor of cytomegalovirus [32], HSV-1, HSV-2, a broad spectrum of genital HPV types, R5, and X4 HIV-1 and was found to exert its action mainly by binding to HS exposed on the cell surface [33e35].…”

Dendrimers

“…58 The additional benefit of this technique is its possible application in complex systems when the interaction of dendriplexes with proteins or glucosaminoglycans is being studied. 59 The basis of the fluorescent dye intercalation assay is the intercalation of fluorescent dyes (ethidium bromide, PicoGreen, GelStar, etc.) into double-stranded DNA or RNA.…”