Light-triggered self-assembled nanostructures for efficient and safe production of engineered therapeutic immune cells

Immunotherapy using chimeric antigen receptor (CAR)-modified cells is a promising approach to treat cancer, which requires the manufacturing of CAR-cell products. An essential part of the manufacturing process is the transfection of patient-derived cells with the CAR construct and with compounds to knock-out immune suppressive pathways to enhance their therapeutic potential. Currently this is performed by viral transduction or by electroporation, which are associated with several drawbacks including safety issues, limited reproducibility, cytotoxicity and reduced therapeutic potency. Photoporation is a new transfection technology that combines laser irradiation with photothermal nanoparticles to safely and efficiently deliver functional molecules into a wide variety of cells. Unfortunately, in spite of its promise it is still not efficient enough for very large gene-editing compounds such as mRNA or pDNA. This project is aimed at enhancing the intracellular delivery of gene-editing compounds in patient-derived T and NK cells by light-triggered ‘nanobombs’ as a new kind of more performant biocompatible photothermal nanomaterial. Different types of light-triggered nanobombs will be synthesized of which the delivery efficiency, acute cytotoxicity, as well as effects on cell homeostasis and therapeutic functionality will be compared with state-of-the art electroporation. The best formulations will be taken forward to produce CAR cells for preclinical e in vivo.