Rapid manufacturing of non-activated potent CAR T cells

Abstract

Chimaeric antigen receptor (CAR) T cells can generate durable clinical responses in B-cell haematologic malignancies. The manufacturing of these T cells typically involves their activation, followed by viral transduction and expansion ex vivo for at least 6 days. However, the activation and expansion of CAR T cells leads to their progressive differentiation and the associated loss of anti-leukaemic activity. Here we show that functional CAR T cells can be generated within 24 hours from T cells derived from peripheral blood without the need for T-cell activation or ex vivo expansion, and that the efficiency of viral transduction in this process is substantially influenced by the formulation of the medium and the surface area-to-volume ratio of the culture vessel. In mouse xenograft models of human leukaemias, the rapidly generated non-activated CAR T cells exhibited higher anti-leukaemic in vivo activity per cell than the corresponding activated CAR T cells produced using the standard protocol. The rapid manufacturing of CAR T cells may reduce production costs and broaden their applicability.

CAR-T Cell Performance: How to Improve Their Persistence?

“…The former protocol for the rapid manufacturing of CAR-T cells may reduce the production costs and broaden their applicability. However, immunosuppressive factors in the TME may hinder the ability to generate functional CAR-T cells using this approach ( 141 ). The facts previously described must be considered relevant in the microenvironment of solid tumors.…”

Manufacturing CD20/CD19-targeted iCasp9 regulatable CAR-TSCMcells usingqCART, theQuantum pBac-based CAR-T system

“…One approach to harness the potency of less-differentiated T cells such as T SCM is to shorten the ex vivo expansion time of CAR-T product. Ghassemi et al (2022) demonstrated that shortening the lentiviral CAR-T manufacturing process to 24hours resulted in a higher percentage of CAR-T cells with a memory phenotype. 32 Though these CAR-T cells displayed stronger in vivo anti-tumor activity than their conventional counterpart, it remains to be seen whether such a rapid manufacturing process can be successfully translated into a clinical setting.…”

“…Ghassemi et al (2022) demonstrated that shortening the lentiviral CAR-T manufacturing process to 24hours resulted in a higher percentage of CAR-T cells with a memory phenotype. 32 Though these CAR-T cells displayed stronger in vivo anti-tumor activity than their conventional counterpart, it remains to be seen whether such a rapid manufacturing process can be successfully translated into a clinical setting. Another approach is to culture CAR-T cells in the presence of specific cytokines or chemical reagents.…”

The Past, Present, and Future of Non-Viral CAR T Cells

“…In recent years, more and more functional advancements are taking place to reduce ex vivo manipulation of CAR T cells in order to preserve their persistence and anti-tumor activity ( 93 ). One of these efforts is represented by the study of Chicaybam and collaborators, demonstrating the possibility of generating SB-engineered anti-CD19 CAR T cells in 8 days activating with Transact (Miltenyi Biotec) after electroporation.…”