Novel cell therapy for the efficient intracellular delivery of genetically encoded payloads 4466

Abstract Description There are currently no approved therapeutic approaches for the targeting of intracellular disease processes using biologics, such as monoclonal antibodies. A major hurdle to this approach is the efficient delivery of biologics across the plasma membrane of the respective target cells. Here, we develop a novel cell therapy allowing the efficient delivery of such payloads using naturally occurring fusogens causing transient membrane-membrane fusion between the engineered cells and target cells. Following identification of fusogens that are amenable to immune cell engineering, by not showing syncytia formation or altering viability, we demonstrate that stable expression of these fusogens can be combined with the co-expression of various payloads, such as DNA-modifying enzymes, single-chain variable fragments, or enzymes. We show that expression of the fusogen is sufficient to allow efficient payload transfer to different target cell types, including epithelial cells and lymphocytes. Using quantitative RT-PCR, we show that mRNA encoding the respective payload is efficiently transferred from the engineered cell to the target cells. Using various model systems, we demonstrate that the transferred payloads are functional in target cells, by demonstrating targeted gene editing using recombinases and base editors, protein degradation, and luciferase activity. This new cell therapy renders acute and chronic intracellular disease processes druggable by genetically encoded therapeutics. Funding Sources Department of Defense CDMRP W81XWH-22-ALSRP-TIA; Maryland Department of Health’s Cigarette Restitution Fund Program CH-649-CRF; NIH P30CA134274 Topic Categories Technological Innovations in Immunology (TECH)