iMATCH - an integrated modular assembly-system for therapeutic combination high-capacity adenovirus gene therapy

Abstract Adenovirus-mediated combination gene therapies have shown promising results in vaccination or treating malignant and genetic diseases. Nevertheless, an efficient system for the rapid assembly and incorporation of therapeutic genes into high-capacity adenoviral vectors (HCAdV) is still missing. Here, we developed the iMATCH platform which enables the generation and production of HCAdVs encoding therapeutic combinations in high quantity and purity within three weeks. Our modular cloning system facilitates the efficient combination of up to four expression cassettes and the rapid integration into HCAdV genomes with defined sizes. Helper viruses (HVs) and purification protocols were optimized to produce HCAdVs with distinct capsid modifications and unprecedented purity (0.1 ppm HVs). The constitution of HCAdVs, with adaptors for targeting and a shield of trimerized scFv for reduced liver clearance, mediated cell- and organ-specific targeting of HCAdVs. As proof of concept, we showed that a single HCAdV encoding an anti PD-1 antibody, IL-12 and IL-2 produced all proteins, and led to tumor regression and prolonged survival in tumor models, comparable to a mixture of single-payload HCAdVs in vitro and in vivo. Therefore, the iMATCH system provides a versatile platform for the generation of high-capacity gene therapy vectors with a high potential for clinical development.