CliniMACS Prodigy Manufacturing of Switchable, AND-Gate CAR T Cells

Research output: Contribution to journalResearch articleContributedpeer-review

Contributors

  • Alexandra von Jutrzenka-Trzebiatowski - , Helmholtz-Zentrum Dresden-Rossendorf (HZDR) (Author)
  • Rutuja Gupte - , Helmholtz-Zentrum Dresden-Rossendorf (HZDR) (Author)
  • Cansu Daglar - , Helmholtz-Zentrum Dresden-Rossendorf (HZDR) (Author)
  • Nicole Berndt - , Helmholtz-Zentrum Dresden-Rossendorf (HZDR) (Author)
  • Claudia Arndt - , Helmholtz-Zentrum Dresden-Rossendorf (HZDR), Mildred Scheel Early Career Center (Author)
  • Michael Bachmann - , National Center for Tumor Diseases Dresden, Helmholtz-Zentrum Dresden-Rossendorf (HZDR), German Cancer Consortium (DKTK) Partner Site Dresden, German Cancer Research Center (DKFZ) (Author)
  • Anja Feldmann - , Helmholtz-Zentrum Dresden-Rossendorf (HZDR), National Center for Tumor Diseases Dresden, German Cancer Consortium (DKTK) Partner Site Dresden, German Cancer Research Center (DKFZ) (Author)

Abstract

The Reverse Chimeric Antigen Receptor (RevCAR) system is an adapter CAR T cell technology that allows the precise tuning of T cell activity and, thus, improved safety management. RevCAR T cells recognize and eradicate tumor cells via a bispecific adapter molecule, termed the RevCAR Target Module (RevTM). To further reduce the risk of on-target off-tumor toxicities, Dual-RevCAR T cells can be employed. These cells harbor two different RevCAR constructs, with the signaling domain of either CD3zeta or CD28. Therefore, Dual-RevCAR T cells only exert their full function when both RevCAR constructs are triggered simultaneously upon recognition of two different tumor antigens via RevTMs, enabling a precise AND-gate targeting approach and rendering them highly interesting for clinical application. For this purpose, standardized and reproducible clinical-grade cell manufacturing is required, for which the CliniMACS Prodigy can be used. Here, we present that automated processing of RevCAR and Dual-RevCAR T cells via the CliniMACS Prodigy results in potent expansion, strong transduction, and a favorable phenotype for clinical application. Moreover, obtained cell products were highly functional in a strict RevTM-dependent manner for both monospecific and AND-gate targeting, clearly underlining their high potential for clinical application against various tumor entities.

Details

Original languageEnglish
Article number5024
JournalInternational journal of molecular sciences
Volume26
Issue number11
Publication statusPublished - 23 May 2025
Peer-reviewedYes

External IDs

PubMedCentral PMC12154027
Scopus 105007726463

Keywords

Keywords

  • Antigens, Neoplasm/immunology, CD28 Antigens/immunology, CD3 Complex/immunology, Humans, Immunotherapy, Adoptive/methods, Neoplasms/therapy, Receptors, Antigen, T-Cell/genetics, Receptors, Chimeric Antigen/immunology, T-Lymphocytes/immunology