Automated manufacture of ΔNPM1 TCR-engineered T cells for AML therapy

Acute myeloid leukemia (AML) is a heterogeneous malignancy that requires further therapeutic improvement, especially for the elderly and subgroups with poor prognosis. A recently discovered T cell receptor (TCR) targeting mutant nucleophosmin 1 (ΔNPM1), presents an attractive option for development of a cancer antigen targeted cellular therapy. Manufacturing of TCR-modified T cells, however, is still limited by a complex, time-consuming and laborious procedure. Therefore, this study specifically addressed the requirements for a scaled manufacture of ΔNPM1-specific T cells in an automated, closed and Good Manufacturing Practice (GMP)-compliant process. Starting from cryopreserved leukapheresis, 2E8 CD8-positive T cells were enriched, activated, lentivirally transduced, expanded and finally formulated. By adjusting and optimizing culture conditions, we additionally reduced the manufacturing time from twelve to eight days while still achieving a clinically relevant yield of up to 5.5E9 ΔNPM1 TCR-engineered T cells. The cellular product mainly consisted of highly viable CD8-positive T cells with an early memory phenotype. ΔNPM1-TCR CD8 T cells manufactured with the optimized process showed specific killing of AML in vitro and in vivo. The process has been implemented in an upcoming phase I/II clinical trial for the treatment of NPM1-mutated AML.