Design and development of a DNA vaccine for high grade serous ovarian cancer

Furthermore, PRAME overexpression correlated with lower metastasis-free survival and was prevalent in platinum-resistant HGSC. A DNA vaccine encoding PRAME was developed using the cationic cell-penetrating peptide RALA to encapsulate and deliver the plasmid DNA (pDNA). The RALA/pPRAME nanoparticles (NPs) were characterized, showing a mean size of <150 nm and a zeta potential >10 at N:P ratios above 4, which is optimal for cellular uptake.

Gel retardation and PicoGreen assays demonstrated effective pDNA condensation and encapsulation efficiency >80% at N:P ratios of 6 and above. In vitro transfection of NCTC 929 and DC 2.4 cell lines confirmed the functionality of the NPs, with observed PRAME overexpression and minimal cytotoxicity (cell viability >80% at N:P ratio 4 and above). Lyophilisation of the RALA/pPRAME NPs with 10% trehalose preserved their characteristics and functionality.

In vivo studies in mice showed an enhanced immune response, with elevated CD4+ and CD8+ T-cell activity, reduced tumor growth, and improved survival rates in vaccinated mice compared to controls. These findings suggest that the RALA/pPRAME vaccine holds promise for treating platinum resistant HGSC and warrants further investigation.<br><br><i>Thesis is embargoed until 31 July 2029</i>.<br><br><br>