DEVELOPMENT AND EVALUATION OF FOLATE-CONJUGATED LIPID NANOPARTICLES FOR THE SITE-SPECIFIC DELIVERY OF SIRNA TARGETING THE KRAS GENE IN NON-SMALL CELL LUNG CANCER (NSCLC)

One of the most exciting and difficult areas of precision oncology is the therapeutic use of RNA interference (RNAi) molecules, such as small interfering RNA (siRNA). Although siRNA may silence almost every gene that causes disease, its clinical translation is significantly hampered by its high rate of systemic breakdown, poor cellular internalisation, and substantial off-target accumulation in the liver and spleen. In order to target Folate Receptor-alpha overexpressing cancers, including non-small cell lung cancer (NSCLC), this study suggests the development and assessment of Ligand-Targeted Nanoparticles (LTNPs) intended for the site-specific delivery of siRNA using Folate-conjugated Lipid Nanoparticles (FA-LNPs) as a model system. LNPs are synthesised using microfluidic mixing as part of the study approach, and an ionisable lipid core is added to aid in endosomal escape. To act as the targeting moiety, folic acid was covalently coupled to the distal ends of PEG-lipids. A homogeneous particle size of about 85 nm with a low Polydispersity Index (PDI < 0.1) was validated by characterisation using Dynamic Light Scattering (DLS), guaranteeing an ideal circulation time. Comparing FR-alpha positive A549 cell lines to non-targeted LNPs, in vitro study showed a 3.5-fold increase in cellular uptake. Additionally, a significant 72% decrease in the expression of the target oncogene (KRAS) at a sub-micromolar concentration was found using quantitative Real-Time PCR (qRT-PCR). These findings suggest that by lowering the necessary dosage and minimising systemic toxicity, ligand-mediated active targeting greatly expands the therapeutic window of RNA therapies. The next generation of personalised RNA therapeutics may depend on the incorporation of cell-specific ligands, according to this research, which offers a scalable framework for the creation of "smart" nanocarriers.