Abstract 3743: New non-integrative MS2-based lentiviral particles for mRNA delivery: A safe and efficient opportunity for gene editing and immunotherapy applications

Safe and efficient cancer therapies using adoptive transfer of engineered T cells or gene editing are very challenging but very promising approaches nowadays. The scientific and clinical communities have been working for a long time together to encounter substantial clinical advances they have made possible thanks to numerous improvements in cell culture and gene transfer methods. Opportunities to improve gene transfer into primary T cells or hematopoietic stem cells involve a better design of the vectors used. Such improvements must lead to an increase of the efficiency in the percentage of positive cells, as well as a better level and duration of expression, cell phenotype preservation and the number of genes delivered. Lentiviral vectors have seen their use largely increased in clinical protocols over the past few years but safety concerns have been highlighted. First, the permanent genetic modification remains a focus of significant regulatory oversight and even integrase- or reverse transcriptase-deficient lentiviral vectors leads to residual integration events. Moreover it has been shown that some resulting CAR T cells can exhibit toxicity due to high and persistent expression. If mRNA delivery is a versatile, flexible, and safe mean for protein therapies, chemical or electroporation-based transfection protocols are known to induce cell toxicity and phenotype modifications of the target cells. Here, we describe a new chimeric lentiviral platform that allows mRNA delivery into the target cells without any genomic signature. The respective properties of the MS2 bacteriophage and the lentiviral vectors have been combined to build a non-integrative packaging system in which the wild type HIV packaging sequence is replaced by the MS2 stem-looprepeats and the MS2 Coat sequence is inserted into the NucleoCapsid sequence. The resulting lentiviral particle is able to deliver a non-viral RNA into the target cell, directly available for protein translation. Transduction of T cells and HSC with these RNA lentiviral particles (RLP) shows an efficient, fast and transient expression of both reporters and functional proteins such as genome editingenzymes. Cell viability of such engineered cells and multiple genes expression analyses will be presented. This transient mRNA delivery mediated by a lentiviral particle is a powerful tool, useful to induce an efficient CAR delivery and ensure a complete loss of the CAR-driven T cells activity. The possibility to express multiple genes at once in the target cells is an attractive therapeutic perspective. One of the advantages of the MS2RLP system is its ability to utilize lentiviral production platforms already validated in clinical settings. The RNAs transferred by the MS2RLPs are directly expressed into the cytoplasm, which completely removes the risk of integration, an important safety consideration for human use. Citation Format: Pascale Bouille, Cedric Auffray, Jean-Christophe Pages, Christine Duthoit, Regis Gayon. New non-integrative MS2-based lentiviral particles for mRNA delivery: A safe and efficient opportunity for gene editing and immunotherapy applications. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3743.