Title:
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A bio-inspired gene therapy approach to treat metastatic prostate cancer
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Five DBVs were synthesized, expressed, purified, and complexed with plasmid DNA for analysis
using a range of physicochemical and biological assays. Four of the vectors (RMHT, RM3GT,
T-RMG, and T-RMR) formed cationic nanoparticles with plasmid DNA, and were capable of
transfecting prostate cancer cells. Conversely RMGT formed highly cytotoxic anionic particles
incapable of transfecting cells. RMHT was the most promising vector as it protected the cargo
from serum and facilitated specific delivery into castrate resistant prostate cancer cells.
RAT, a fusogenic peptide, is composed of an arginine rich cell penetrating peptide, a steric
alpha-helicallinker, and a metastatic tumour targeting ligand named TMTP1. RAT formed
non-toxic cationic nanoparticles with plasmid DNA that were capable of specifically transfecting
castrate resistant prostate cancer cells through receptor-mediated endocytosis. Furthermore,
RAT nanoparticies remained stable over a wide temperature range, and protected DNA from
degradation by serum endonucleases.
RAT was investigated in combination with iNOS plasmids driven by constitutive (CMV) and tumour
type specific (hOC) promoters. RAT facilitated iNOS gene expression, and subsequent
generation of μM concentrations of the NO free radical. This equated to a maximum of 59% cell
kill and less than 74% clonogenicity in prostate cancer cells. Systemic delivery in vivo slightly
delayed tumour growth, and therefore future considerations will include dosing strategies ,and
circulation enhancement. These bio-inspired gene delivery platforms overcome a multitude of
biological barriers and the results generated in this thesis warrant further investigation.
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