Generation and GMP scale-up of human CAR-T cells using non-viral <i>Sleeping Beauty</i> transposons for B cell malignances.

ABSTRACT: Most CAR-T therapies rely on genetic T cell engineering with integrating viral vectors that, although effective, are associated with prohibitive costs. Here we have generated TranspoCART19 cells, a fully functional 4-1BB second-generation CAR-T cell product targeting CD19, fused to a truncated version of the human EGFR (hEGFRt) as reporter gene and safety switch, based on the Sleeping Beauty transposon delivery system. Our manufacturing protocol allowed generation of TranspoCART19 cells under GMP conditions, showing similar in vitro and in vivo antitumoral efficacy than conventional CAR-T cells generated with lentiviral vectors. Additionally, membrane expression of hEGFRt facilitated in vivo CAR-T cell elimination after cetuximab administration. Safety analyses

SUBMITTER: Diez B 

PROVIDER: S-EPMC11874549 | biostudies-literature | 2025 Mar

REPOSITORIES: biostudies-literature