mRNA-based CAR-T Cell Engineering via Antibody-Conjugated Lipid Nanoparticles

Abstract

This thesis investigates the use of lipid nanoparticles (LNPs) for chimeric antigen receptor (CAR) mRNA (messenger RNA) delivery to T cells as a non-viral approach for cell engineering. Experiments assessed LNP uptake across different cell types and demonstrated minimal uptake in T cells. To address this limitation, antibody-conjugated LNPs were used to enable efficient delivery to T cells. Transfection efficiency was evaluated using GFP reporter mRNA as well as CAR expression detected via an FMC63 idiotype antibody. Functional activity was assessed through co-culture killing assays against CD19-positive Nalm6-GFP target cells across multiple effector-to-target (E:T) ratios. The results demonstrate antibody-conjugated LNPs enhance mRNA delivery to T cells and enable the generation of functional CAR-T cells capable of mediating tumour cell killing.