CAREER: Novel expansion strategies for biomanufacturing of natural killer cells

工程T细胞疗法最近被美国食品和药物管理局批准用于对抗癌症。这些疗法利用人体的免疫系统更有效地攻击晚期癌症。自然杀伤（NK）细胞是用于治疗血癌和实体瘤的有前景的免疫细胞群体。需要安全、大规模的NK细胞生产来有效治疗大量患者。免疫细胞是复杂的，因此很难大量生产NK细胞。确保每个细胞具有高水平的活性增加了治疗性细胞制造的复杂性。一个挑战是开发一种不需要引入其他细胞来激活NK细胞的方法。研究人员建议开发一种能够激活NK细胞的人工脂质囊泡。这种方法将可用于生产各种治疗性细胞。还计划开展教育和外联活动。女性和代表性不足的少数民族将被积极招募，以追求STEM职业。这将得到视频和其他课程材料的支持，以提高数学能力。目前，NK细胞的大规模生长需要人工抗原呈递细胞（aAPC）的多轮刺激。它们最常来源于K562白血病细胞系。这些饲养细胞的使用导致临床生物制造过程中的监管问题，并阻碍治疗采用。该项目将开发用于大规模NK细胞扩增的无饲养层方法。通过阐明与aAPC共培养刺激NK细胞的机制，可以开发用于在没有细胞aAPC的情况下扩增NK细胞的方法。将在多轮刺激期间表征NK代谢。理解aAPC刺激如何诱导NK生长是特别感兴趣的。将在每轮刺激之前和之后进行RNA-seq分析，以鉴定被调节的通路。这些结果将为设计合成磷脂囊泡提供信息，这些囊泡向NK细胞提供关键的激活信号。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

Engineered T cell therapies were recently approved by the Food and Drug Administration to fight cancer. These therapies harness the body's immune system to more effectively attack advanced stage cancer. Natural killer (NK) cells are a promising immune cell population for the treatment of blood cancers and solid tumors. Safe, large-scale NK cell production is needed to effectively treat large numbers of patients. Immune cells are complex, so producing NK cells in large quantities is difficult. Guaranteeing that each cell has a high level of activity adds a level of complexity to therapeutic cell manufacturing. One challenge is to develop a process that does not require the introduction of other cells to activate the NK cells. The investigators propose to develop an artificial lipid vesicle that will activate the NK cells. This approach will be useful in the production of a variety of therapeutic cells. Education and outreach activities are also planned. Women and underrepresented minorities will be actively recruited to pursue STEM careers. This will be supported by videos and other curricular material developed to bolster competency in mathematics.Large-scale growth of NK cells currently requires multiple rounds of stimulation with artificial antigen-presenting cells (aAPCs). These are most often derived from the K562 leukemia cell line. The use of these feeder cells leads to regulatory concerns in clinical biomanufacturing processes and hinders therapeutic adoption. This project will develop feeder-free approaches for large-scale NK cell expansion. By elucidating the mechanisms through which co-culture with aAPCs stimulates NK cells, processes for expansion of NK cells without cellular aAPCs can be developed. NK metabolism will be characterized during multiple rounds of stimulation. Understanding how aAPC stimulation induces NK growth is of particular interest. RNA-seq analysis will be performed before and after each round of stimulation to identify pathways that are modulated. These results will inform the design of synthetic phospholipid vesicles that present critical activating signals to the NK cells.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.