Highly Efficient And Robust Delivery Of Multicistronic Therapeutic Cargo In Immune Cells For Both RUO And Clinical Applications

By Kirsti L. Walker, Bryan J. Jones, Kalyani Kota, Katie Hornberger, Kari T. Wenzel, John R. Leerar, Dalton Smedley, Rebecca Haugen, Ariel Miller, Joseph G. Skeate, Emily J. Pomeroy, Nicholas J. Slipek, Bryce J. Wick, Walker S. Lahr, Erin Stelljes, Beau R. Weber, Brandon S. Moriarty, David L. Hermanson, Xiaobai Patrinostro, Neil Otto

The development of genome engineering tools has led to various clinical trials for immune and stem cell therapies. Most of these therapies use viral vectors for delivery, which carry risks and limitations. To overcome such challenges, a non-viral transposase-based editing platform called TcBuster™-M (TcB-M) has been developed. TcB-M provides rapid cell manufacturing at lower cost and is less constrained by cargo size. It can be combined with endonucleases such as CRISPR for more complex editing.

The platform shows higher cargo integration rates in primary T-cells and peripheral blood-derived NK cells, without impacting cell growth. TcB-M has a safer integration profile compared to other engineered CAR-Ts and can accelerate the delivery of crucial therapies. The tool also outperforms other transposases in terms of transposition efficiency and has been used in vivo to control the tumor burden of Burkitt’s lymphoma. The study also includes analyses of site-integration, transposition efficiency, and the combination of CAR- T and CAR- NK therapy in controlling tumors.