CAREER: Novel expansion strategies for biomanufacturing of natural killer cells

转基因T细胞疗法最近被美国食品和药物管理局批准用于抗癌。这些疗法利用人体的免疫系统来更有效地攻击晚期癌症。自然杀伤(NK)细胞是治疗血癌和实体瘤的一种很有前途的免疫细胞群。需要安全、大规模的NK细胞生产才能有效地治疗大量患者。免疫细胞很复杂，因此很难大量生产NK细胞。保证每个细胞具有高水平的活性增加了治疗细胞制造的复杂性。其中一个挑战是开发一种不需要引入其他细胞来激活NK细胞的过程。研究人员提议开发一种能够激活NK细胞的人造脂囊。这种方法将在生产各种治疗性细胞方面有用。还计划开展教育和外联活动。将积极招募妇女和任职人数不足的少数群体从事STEM职业。这将得到为提高数学能力而开发的视频和其他课程材料的支持。目前，NK细胞的大规模增长需要人工抗原呈递细胞(AAPC)的多轮刺激。这些细胞最常来自K562白血病细胞系。这些饲养层细胞的使用导致了临床生物制造过程中的监管问题，并阻碍了治疗的采用。该项目将开发无饲养器的方法来大规模扩增NK细胞。通过阐明与aAPC共培养刺激NK细胞的机制，可以开发在没有细胞aAPC的情况下扩增NK细胞的过程。NK代谢将在多轮刺激中表现出来。了解AAPC刺激如何诱导NK生长是特别有趣的。将在每一轮刺激前后进行RNA-SEQ分析，以确定被调制的通路。这些结果将为向NK细胞提供关键激活信号的合成磷脂微囊的设计提供参考。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势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.