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Anti-CD19 CAR-T cell immunotherapy for treatment of cancer is showing promising results in in patients harboring B cell malignancies, with overall response rates of 73%. However, there is no standard protocol for the generation of CAR-modified T cells, with different genetic modification vectors and expansion protocols being used. Viral vectors insert the transgene in the genome of the cell, providing long term CAR expression. However, their use is associated with high production costs and cumbersome quality controls, impacting the final cost of CAR-T cell therapies. Sleeping Beauty (SB) transposon system consist of plasmidbased integrative vectors that, through a cut-and-paste mechanism catalyzed by a transposase, recognize inverted terminal repeats flanking the transgene and insert it in the target cell genome. We show here that this system, combined with LCL-based T cell expansion can be used to efficiently transfect primary T lymphocytes and induce long term CAR expression throughout T cell expansion.

CAR-T cells generated using Sleeping Beauty vectors and expanded with a lymphoblastoid cell line (LCL) display in vivo antitumor activity