Supramolecular Assembly of Amino Acid Based Cationic Polymer for Efficient Gene Transfection Efficiency in Triple Negative Breast Cancer.

Abstract

The success of gene therapy is enormously dependent on an efficient gene carrier, and in this context, cationic polymers still continue to play a major role particularly with respect to the safety issue compared to viral vectors. Developing an efficient gene carrier system having promising gene transfection efficiency with low toxicity is the foremost impediment associated with a nonviral carrier. Here, we explored amino acid based biocompatible polymers synthesized via reversible addition-fragmentation chain transfer (RAFT) polymerization where glycine (Gly), leucine (Leu), and phenyl alanine (Phe) amino acids were used as the pendent groups of the polymeric brushes. The presence of both a hydrophobic group (long chain aliphatic group associated with the RAFT agent) and hydrophilic amino groups was associated with the supramolecular assembly of the polymeric chain having hydrodynamic sizes within the range of 150-300 nm with a positive zeta potential of 30 ± 5 mV. All polymers showed very low toxicity and possessed >80% cell viability even at a very high concentration of 1000 μg/mL against both normal and cancerous cells. In addition to this, the polymers also showed excellent blood compatibility, and negligible hemolysis was observed at the concentration of 500 μg/mL. All polymers showed efficient DNA complexation capability as well as excellent protection of DNA against highly negatively charged surfactant and enzymatic digestion, although the efficiency was dependent on the N/P ratio of polymer/DNA complexes. Interestingly, the phenyl alanine moiety containing polymer brush P(HEMA-Phe-NH2) showed a hexagonal shaped nanoparticle after complexation with pDNA and consequently showed higher cellular uptake, resulting in a higher transfection efficiency in a triple negative breast cancer cell, the MDA-MB-231 cell. Therefore, the synthesized polymer containing an amino acid pendent group, especially the phenyl alanine moiety, may be a promising nonviral gene carrier system in gene therapy application in the future.