Pfefferle Aline, Contet Julian, Wong Kahlia, Chen Charlotte, Verhoeyen Els, Slichter Chloe K, Schluns Kimberly S, Cursons Joseph, Berry Richard, Nikolic Iva, Rautela Jai, Huntington Nicholas D
Biomedicine Discovery Institute and the Department of Biochemistry and Molecular Biology Monash University Clayton VIC Australia.
oNKo-Innate Pty Ltd Moonee Ponds VIC Australia.
Clin Transl Immunology. 2024 May 2;13(5):e1507. doi: 10.1002/cti2.1507. eCollection 2024.
Autologous chimeric antigen receptor (CAR) T-cell therapy of B-cell malignancies achieves long-term disease remission in a high fraction of patients and has triggered intense research into translating this successful approach into additional cancer types. However, the complex logistics involved in autologous CAR-T manufacturing, the compromised fitness of patient-derived T cells, the high rates of serious toxicities and the overall cost involved with product manufacturing and hospitalisation have driven innovation to overcome such hurdles. One alternative approach is the use of allogeneic natural killer (NK) cells as a source for CAR-NK cell therapy. However, this source has traditionally faced numerous manufacturing challenges.
To address this, we have developed an optimised expansion and transduction protocol for primary human NK cells primed for manufacturing scaling and clinical evaluation. We have performed an in-depth comparison of primary human NK cell sources as a starting material by characterising their phenotype, functionality, expansion potential and transduction efficiency at crucial timepoints of our CAR-NK manufacturing pipeline.
We identified adult peripheral blood-derived NK cells to be the superior source for generating a CAR-NK cell product because of a higher maximum yield of CAR-expressing NK cells combined with potent natural, as well as CAR-mediated anti-tumor effector functions.
Our optimised manufacturing pipeline dramatically improves lentiviral transduction efficiency of primary human NK cells. We conclude that the exponential expansion pre- and post-transduction and high on-target cytotoxicity make peripheral blood-derived NK cells a feasible and attractive CAR-NK cell product for clinical utility.
B 细胞恶性肿瘤的自体嵌合抗原受体（CAR）T 细胞疗法使很大一部分患者实现了长期疾病缓解，并引发了将这一成功方法应用于其他癌症类型的深入研究。然而，自体 CAR-T 细胞制造涉及的复杂物流、患者来源 T 细胞的适应性受损、严重毒性的高发生率以及产品制造和住院的总体成本推动了创新以克服这些障碍。一种替代方法是使用同种异体自然杀伤（NK）细胞作为 CAR-NK 细胞疗法的来源。然而，传统上这种来源面临着众多制造挑战。
为了解决这一问题，我们开发了一种优化的扩增和转导方案，用于为扩大制造规模和临床评估准备的原代人 NK 细胞。我们通过在 CAR-NK 细胞制造流程的关键时间点表征其表型、功能、扩增潜力和转导效率，对作为起始材料的原代人 NK 细胞来源进行了深入比较。
我们确定成人外周血来源的 NK 细胞是生成 CAR-NK 细胞产品的优质来源，因为表达 CAR 的 NK 细胞最大产量更高，同时具有强大的天然以及 CAR 介导的抗肿瘤效应功能。
我们优化的制造流程显著提高了原代人 NK 细胞的慢病毒转导效率。我们得出结论，转导前后的指数扩增和高靶向细胞毒性使外周血来源的 NK 细胞成为一种用于临床的可行且有吸引力的 CAR-NK 细胞产品。
