Wang Chelsia Qiuxia, Leong Leonard, Yang Lei, Xu Shengli, Lam Kong-Peng, Choo Andre Boon-Hwa, Tan Andy Hee-Meng
Immune Cell Manufacturing, Bioprocessing Technology Institute (BTI), Agency for Science, Technology and Research (ASTAR), Singapore, Singapore.
Biotherapeutics Development, Bioprocessing Technology Institute (BTI), Agency for Science, Technology and Research (ASTAR), Singapore, Singapore.
Front Immunol. 2025 Apr 3;16:1557766. doi: 10.3389/fimmu.2025.1557766. eCollection 2025.
Chimeric antigen receptor (CAR)-modified cell therapy products approved for clinical treatment of hematological malignancies have hitherto been based on T cells. NK cells represent a promising immune cell type that can be considered for CAR engineering due to their potential to be generated as off-the-shelf allogeneic cellular therapy. Viral transduction of NK cells with CARs has been fraught with challenges of long process time and poor CAR transduction efficiency. Here, we describe the development of an optimized protocol for electroporation-based delivery of CAR mRNA into NK cells expanded from human peripheral blood mononuclear cells in the presence of co-stimulating feeder cells. This enabled rapid assessment of the functional capacity of NK cells transiently expressing various CARs to kill liquid and solid tumor cells . Ultimately, we anticipate that such an approach will enable selection of CAR candidates for their subsequent clinical applicability and manufacturability.
迄今为止，已获批用于临床治疗血液系统恶性肿瘤的嵌合抗原受体（CAR）修饰的细胞治疗产品均基于T细胞。自然杀伤（NK）细胞是一种很有前景的免疫细胞类型，由于其有潜力作为现成的异基因细胞疗法产生，因此可考虑用于CAR工程。用CAR对NK细胞进行病毒转导一直充满了处理时间长和CAR转导效率低的挑战。在此，我们描述了一种优化方案的开发，该方案用于在共刺激饲养细胞存在的情况下，基于电穿孔将CAR mRNA递送至从人外周血单个核细胞扩增而来的NK细胞中。这使得能够快速评估瞬时表达各种CAR的NK细胞杀伤液体和实体肿瘤细胞的功能能力。最终，我们预计这种方法将能够选择CAR候选物以用于其后续的临床适用性和可制造性。
