Abstract
Background & Aim The approval of the very first chimeric antigen receptor (CAR) - T therapies (Kymriah and Yescarta) by European and American agencies has marked a truly revolutionary period in cancer immuno-therapeutics. Currently, most autologous CAR-T therapies are generated via a centralized manufacturing process whereby the patients’ T cells are isolated, followed by introduction of the genetic material via (lenti) viral transduction, expansion of the transduced T cells and finally re-infusion of the CAR-T cells into the patient. The generation of autologous CAR-T treatments is very laborious and is associated with very high costs. Additionally, the requirement of large batches of clinical grade (lenti) virus is adding further complexity to the overall process. For that reason, efforts are being made to both simplify the manufacturing process and reduce the overall costs. The work described herein is part of a consolidated effort among leading UK universities to generate an on- or near- body manufacturing device that will generate autologous CAR-T cells. The specific primary aim of this study was to identify the optimal methodology for CAR gene delivery within a micro factory setting. Methods, Results & Conclusion The gene delivery methods that were explored were both genome-integrative and non-integrative, were both viral and non-viral and included: 1) lentiviral vector delivery, 2) mRNA delivery and 3) minicircle-based Sleeping Beautytransposon/transposase delivery. The efficiency of CAR-T cell generation was assessed via flow cytometric analysis.
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