Yonezawa Ogusuku Isabella Elias, Herbel Vera, Lennartz Simon, Brandes Caroline, Argiro Eva, Fabian Caroline, Hauck Carola, Hoogstraten Conny, Veld Sabrina, Hageman Lois, Teppert Karin, Koutsoumpli Georgia, Griffioen Marieke, Mockel-Tenbrinck Nadine, Schaser Thomas, de Groot Rosa, Johnston Ian C D, Lock Dominik
Miltenyi Biotec B.V. & Co. KG, 51429 Bergisch Gladbach, Germany.
Department of Hematology, Leiden University Medical Center, 2333 ZA Leiden, the Netherlands.
Mol Ther Methods Clin Dev. 2024 Feb 27;32(2):101224. doi: 10.1016/j.omtm.2024.101224. eCollection 2024 Jun 13.
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 T cell 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 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-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 12 to 8 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 and . The process has been implemented in an upcoming phase 1/2 clinical trial for the treatment of NPM1-mutated AML.
急性髓系白血病（AML）是一种异质性恶性肿瘤，需要进一步改进治疗方法，尤其是针对老年患者和预后不良的亚组患者。最近发现的靶向突变型核磷蛋白1（ΔNPM1）的T细胞受体（TCR）为开发癌症抗原靶向细胞疗法提供了一个有吸引力的选择。然而，TCR修饰的T细胞的制造仍然受到复杂、耗时且费力的程序的限制。因此，本研究专门探讨了在自动化、封闭且符合药品生产质量管理规范（GMP）的过程中大规模制造ΔNPM1特异性T细胞的要求。从冷冻保存的白细胞单采开始，富集、激活、慢病毒转导、扩增2E8 CD8阳性T细胞，最后进行制剂配制。通过调整和优化培养条件，我们还将制造时间从12天缩短至8天，同时仍能实现高达5.5E9个经ΔNPM1 TCR工程改造的T细胞的临床相关产量。细胞产品主要由具有早期记忆表型的高活力CD8阳性T细胞组成。采用优化工艺制造的ΔNPM1 TCR CD8 T细胞对AML显示出特异性杀伤作用。该工艺已在即将开展的1/2期临床试验中用于治疗NPM1突变的AML。
