Using Piggybac transposons, scientists develop a simple and cost-effective protocol for manufacturing chimeric antigen receptor (CAR) targeting CD19 for adoptive therapies.
Adoptive immunotherapy using CAR-T cells targeting CD19 has been tested as a novel approach for the treatment of various cancers. Current clinical trials use either retroviral or lentiviral-mediated gene transfer for engineering CAR-T cells. However, use of these viral-based vectors for T-cell modification represents significant manufacturing challenge due to the complexity of the transduction approach and the need for thorough quality control.
To save the labor, reduce the cost and improve the safety of CAR-modified T-cell therapy, Dr Pavel Otáhal and colleagues at Prague’s Institute of Hematology and Blood Transfusion developed a protocol for non-viral gene-transfer using piggyBac-transposon system. In the study published in Cytotherapy, the group transferred CAR19 gene into T cells using piggyBac-transposon system by electroporation and cultivated the cells in the presence of cytokines IL-4, IL-7 and IL-21 for selective expansion of CAR19-T cells.
The protocol resulted in >90% CAR+ T cells, which was observed following activation of the CAR receptor by either its cognate ligand (i.e., CD19 expressed on the surface of B cells) or anti-CAR antibody, and cultivation in the presence of cytokines IL-4 and IL-7.
Thus, the study presents PiggyBac-mediated CD19-specific T-cell therapy as a simple and inexpensive method for adoptive immunotherapy against cancers.
Source: Ptáčková P et al., A new approach to CAR T-cell gene engineering and cultivation using piggyBac transposon in the presence of IL-4, IL-7 and IL-21. Cytotherapy, February 2018. Reference