EAGER: Biomanufacturing: Towards Reproducible and Scalable Biomanufacturing of Tumor-Specific T Cells with Optimal Phenotype and Function for Personalized Immunotherapy

在美国，约25%的死亡是由癌症造成的。开发一种有效的过继t细胞疗法（ACT），一种有前途的高度个性化的癌症免疫疗法，对社会有重大的好处。然而，开发可重复和可扩展的制造工艺来可靠地产生具有高抗肿瘤活性的T细胞仍然是一个重大挑战。该项目旨在通过开发一种快速、简单、全面的t细胞分析方法来解决这一挑战。它代表了一种全新的、完全不同的方法，具有变革潜力，解决了ACT商业化的多重障碍，包括制造可重复性、治疗有效性和患者之间的可变性。研究成果还将用于加强本科和研究生课程，推动工程推广工作，鼓励K-12学生尽早接触科学和工程，并提高公众的科学素养。创新的教学方法将被开发出来，可以很容易地被各级教育工作者采纳，以改善学生的学习。这些努力将共同帮助培养具有工程和生命科学技能和基础知识的新一代工程师。过继性T细胞疗法（ACT）是一种高度个性化的癌症免疫疗法，它涉及将体外制造的自然或基因工程肿瘤反应性T细胞输注到患者体内。单个naïve人类T细胞的扩增产生异质群体，表现出一系列表型和功能。虽然这种多样性有助于T细胞在不同阶段获得针对不同类型病原体和癌症的特化反应，但它代表了用于治疗目的的可复制、大规模制造T细胞的重大技术挑战。因此，建立一种高效的t细胞分析方法来密切监测它们的表型、功能和特异性是至关重要的。不幸的是，目前的标准技术需要多次样品运行，增加了制造过程的复杂性、可变性和工作量。为了解决这个问题，将开发一种T细胞分析方法，以便在单次运行中同时检测异质群体中单个T细胞的数十个参数。将开发所产生的高维数据的可视化方法，以促进制造过程质量控制步骤中的客观数据分析自动化。同时，通过开发和应用一种创新的人工抗原呈递系统，针对乳腺癌中发现的一系列亚显性肿瘤相关抗原的T细胞将从人类供体中扩增出来。结合这里开发的T细胞谱分析方法，这些T细胞为系统筛选亚显性T细胞的最佳扩增条件和确定与最佳抗肿瘤活性相关的分子指标提供了宝贵的机会。这些指标的定义将进一步简化质量控制协议，并促进ACT制造过程的标准化。

1645229 - Wen Cancer is responsible for about 25% of deaths in the US. Developing an effective adoptive T-cell therapy (ACT), a promising and highly personalized cancer immunotherapy, holds significant benefits for society. However, it remains a significant challenge to develop reproducible and scalable manufacturing processes to reliably generate T cells with high anti-tumor activity. This project aims to address this challenge by developing a rapid, simple, and comprehensive T-cell profiling method. It represents a novel and radically different approach with transformative potential by addressing multiple roadblocks to the commercialization of ACT, including manufacturing reproducibility, therapeutic effectiveness and patient-to-patient variability. Research findings will also be used to strengthen undergraduate and graduate curricula, to power engineering outreach endeavors that encourage early exposure of K-12 students to science and engineering, and to increase public scientific literacy. Innovative pedagogies will be developed that can be readily incorporated by educators at all levels to improve their students' learning. These efforts will collectively help create a new generation of engineers with skills and fundamental knowledge in both engineering and life sciences. Adoptive T-cell therapy (ACT) is a highly personalized cancer immunotherapy that involves the infusion into patients of their natural or genetically engineered tumor-reactive T cells manufactured ex vivo. Expansion of a single naïve human T cell yields a heterogeneous population exhibiting a range of phenotype and function. While this diversity helps T cells acquire specialized responses to different types of pathogens and cancers at different stages, it represents a significant technical challenge to reproducible, large-scale manufacturing of T cells for therapeutic purposes. Therefore, it is critical to have a high performance T-cell profiling method to closely monitor their phenotype, function and specificity. Unfortunately, the current standard technology requires several sample runs, adding complexity, variability and workload to the manufacturing process. To address this issue, a T-cell profiling method will be developed to enable simultaneous detection of dozens of parameters on single T cells in a heterogeneous population in a single run. Visualization methods of the resulting high-dimensional data will be developed to facilitate objective data-analysis automation in the quality control steps of the manufacturing process. In parallel, by developing and applying an innovative artificial antigen presentation system, T cells specific to a range of subdominant tumor-associated antigens found in breast cancer will be expanded from human donors. Coupled with the T-cell profiling method developed here, these T cells provide valuable opportunities to systematically screen for optimal expansion conditions of subdominant T cells and define their molecular metrics that correlate with the best anti-tumor activity. The definition of such metrics will further simplify the quality control protocol and facilitate the standardization of the ACT manufacturing process.