NK细胞疗法作为一种潜在的癌症治疗方法已经获得了极大的关注。走向治疗用途,NK细胞需要被激活和扩增以获得高效和大量的有效剂量。这通常通过用细胞因子离体刺激以增强功能或扩增10-14天以增加其活性和数量来完成。对于现成的产品,获得产生大剂量的有效NK细胞的稳健方法是非常期望的。值得注意的是,过去的报道表明,用IL-12,IL-15和IL-18刺激NK细胞赋予它们类似记忆的特性,更好的抗肿瘤活性,和坚持。虽然这种方法产生的NK细胞具有临床上有利的特征,并得到了令人鼓舞的早期血液恶性肿瘤治疗结果的支持。其有限的可扩展性,初始剂量的可变性,以及患者特定生产的必要性阻碍了其更广泛的应用。在这项研究中,用来自K562-41BBL-mbIL21细胞的PM21颗粒刺激NK细胞与使用细胞因子IL-12,IL-15和IL-18的记忆样诱导相结合,以产生具有增强的抗肿瘤功能的NK细胞。使用细胞因子与PM21颗粒(细胞因子和颗粒,CAP)显着增强NK细胞扩增,在14天内实现了惊人的8200倍。机械上,与单独的PM21颗粒相比,这种显着改善是由于关键刺激配体(4-1BBL和IL-2)的受体上调,导致驱动NK细胞大量生长的协同作用,展示了更有效的治疗应用的潜力。CAP-NK细胞的治疗潜力由增强的代谢适应性证明。持久性,和抗肿瘤功能在体外和体内。最后,CAP-NK细胞适用于目前用于开发治疗性NK细胞产品的技术,包括基于CRISPR/Cas9的技术来产生三基因敲除或基因敲入。一起来看,这些数据表明,细胞因子的添加增强了已经有效的体外产生治疗性NK细胞与PM21颗粒的方法,产生优异的NK细胞产品的制造效率和潜在的治疗应用。
NK cell therapeutics have gained significant attention as a potential cancer treatment. Towards therapeutic use, NK cells need to be activated and expanded to attain high potency and large quantities for an effective dosage. This is typically done by ex vivo stimulation with cytokines to enhance functionality or expansion for 10-14 days to increase both their activity and quantity. Attaining a robust methodology to produce large doses of potent NK cells for an off-the-shelf product is highly desirable. Notably, past reports have shown that stimulating NK cells with IL-12, IL-15, and IL-18 endows them with memory-like properties, better anti-tumor activity, and persistence. While this approach produces NK cells with clinically favorable characteristics supported by encouraging early results for the treatment of hematological malignancies, its limited scalability, variability in initial doses, and the necessity for patient-specific production hinder its broader application. In this study, stimulation of NK cells with PM21-particles derived from K562-41BBL-mbIL21 cells was combined with memory-like induction using cytokines IL-12, IL-15, and IL-18 to produce NK cells with enhanced anti-tumor function. The use of cytokines combined with PM21-particles (cytokine and particle, CAP) significantly enhanced NK cell expansion, achieving a remarkable 8,200-fold in 14 days. Mechanistically, this significant improvement over expansion with PM21-particles alone was due to the upregulation of receptors for key stimulating ligands (4-1BBL and IL-2), resulting in a synergy that drives substantial NK cell growth, showcasing the potential for more effective therapeutic applications. The therapeutic potential of CAP-NK cells was demonstrated by the enhanced metabolic fitness, persistence, and anti-tumor function both in vitro and in vivo. Finally, CAP-NK cells were amenable to current technologies used in developing therapeutic NK cell products, including CRISPR/Cas9-based techniques to generate a triple-gene knockout or a gene knock-in. Taken together, these data demonstrate that the addition of cytokines enhanced the already effective method of ex vivo generation of therapeutic NK cells with PM21-particles, yielding a superior NK cell product for manufacturing efficiency and potential therapeutic applications.