Niraj Shrestha, Michael J Dee, Pallavi Chaturvedi, Gilles M Leclerc, Mary Mathyer, Celeste Dufour, Laura Arthur, Michelle Becker-Hapak, Mark Foster, Ethan McClain, Natalia Valderrama Pena, Karen Kage, Xiaoyun Zhu, Varghese George, Bai Liu, Jack Egan, Christian Echeverri, Meng Wang, Lijing You, Lin Kong, Liying Li, Melissa M Berrien-Elliott, Matthew L Cooper, Todd A Fehniger, Peter R Rhode, Hing C Wong
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引用次数: 0
摘要
使用IL-12、IL-15和IL-18在体外过夜激活产生的记忆样(ML)自然杀伤(NK)细胞进行的适应性细胞疗法(ACT)已显示出治疗血液恶性肿瘤的前景。我们最近报道了一种由 IL-12、IL-15 和 IL-18 结构域组成的多功能融合分子 HCW9201,它可以取代单个细胞因子,用于人类 ML NK 细胞编程的启动("启动 "步骤)。然而,这种方法不包括体内外扩增,因此限制了测试不同剂量和计划的能力。在此,我们报告了由人类 IL-7、IL-15 和 IL-21 细胞因子组成的多功能融合分子 HCW9206 的设计和生成。我们观察到,用 HCW9206 和 HCW9101(一种 IgG1 抗体,可识别 HCW9206 的支架结构域)培养 14 天的 HCW9201 激发的人 NK 细胞扩增了 300 倍以上("扩增 "步骤)。这种扩增既依赖于 HCW9206 细胞因子,也依赖于 IgG1 mAb 与 NK 细胞上 CD16 受体的相互作用。由此产生的 "Prime and Expand" ML NK 细胞表现出更高的代谢能力、稳定的表观遗传 IFNG 启动子去甲基化、更强的体外和体内抗肿瘤活性,以及在 NSG 小鼠体内更强的持久性。因此,"Prime and Expand "策略代表了一种简单的无饲养细胞方法,可简化临床级 ML NK 细胞的生产,支持多剂量和现成的 ACT。
A "Prime and Expand" strategy using the multifunctional fusion proteins to generate memory-like NK cells for cell therapy.
Adoptive cellular therapy (ACT) using memory-like (ML) natural killer (NK) cells, generated through overnight ex vivo activation with IL-12, IL-15, and IL-18, has shown promise for treating hematologic malignancies. We recently reported that a multifunctional fusion molecule, HCW9201, comprising IL-12, IL-15, and IL-18 domains could replace individual cytokines for priming human ML NK cell programming ("Prime" step). However, this approach does not include ex vivo expansion, thereby limiting the ability to test different doses and schedules. Here, we report the design and generation of a multifunctional fusion molecule, HCW9206, consisting of human IL-7, IL-15, and IL-21 cytokines. We observed > 300-fold expansion for HCW9201-primed human NK cells cultured for 14 days with HCW9206 and HCW9101, an IgG1 antibody, recognizing the scaffold domain of HCW9206 ("Expand" step). This expansion was dependent on both HCW9206 cytokines and interactions of the IgG1 mAb with CD16 receptors on NK cells. The resulting "Prime and Expand" ML NK cells exhibited elevated metabolic capacity, stable epigenetic IFNG promoter demethylation, enhanced antitumor activity in vitro and in vivo, and superior persistence in NSG mice. Thus, the "Prime and Expand" strategy represents a simple feeder cell-free approach to streamline manufacturing of clinical-grade ML NK cells to support multidose and off-the-shelf ACT.
期刊介绍:
Cancer Immunology, Immunotherapy has the basic aim of keeping readers informed of the latest research results in the fields of oncology and immunology. As knowledge expands, the scope of the journal has broadened to include more of the progress being made in the areas of biology concerned with biological response modifiers. This helps keep readers up to date on the latest advances in our understanding of tumor-host interactions.
The journal publishes short editorials including "position papers," general reviews, original articles, and short communications, providing a forum for the most current experimental and clinical advances in tumor immunology.