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

Abstract

Acute myeloid leukemia (AML) is a heterogeneous malignancy that requires further therapeutic improvement, especially for the elderly and for subgroups with poor prognosis. A recently discovered Tcell receptor (TCR) targeting mutant nucleophosmin 1 (ΔNPM1) presents an attractive option for the development of a cancer antigen-targeted cellular therapy. Manufacturing of TCR-modified Tcells, 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 Tcells in an automated, closed, and good manufacturing practice-compliant process. Starting from cryopreserved leukapheresis, 2E8 CD8-positive Tcells were enriched, activated, lentivirally transduced, expanded, and finally formulated. By adjusting and optimizing culture conditions, we additionally reduced the manufacturing time from 12 to 8days while still achieving a clinically relevant yield of up to 5.5E9 ΔNPM1 TCR-engineered Tcells. The cellular product mainly consisted of highly viable CD8-positive Tcells with an early memory phenotype. ΔNPM1 TCR CD8 Tcells manufactured with the optimized process showed specific killing of AML invitro and invivo. The process has been implemented in an upcoming phase 1/2 clinical trial for the treatment of NPM1-mutated AML.