Single-cell dynamic analysis to improve the biomanufacturing process for engineering of cell therapy products

免疫细胞可以成为多种疾病有效治疗的基础。生产这些具有均匀活性的细胞是困难的。本项目将重点开发一种能够从无效细胞中分选有效细胞的微流体装置。有效人口将被表征。期望获得的见解将有助于设计更有效的嵌合抗原受体（CAR） T细胞。针对高中学生的互动讲座和实践活动将是该项目支持的外展活动的关键组成部分。通过多样性和女性参与工程项目激发高中女生对未来生物技术和生物工程研究的兴趣，应该会增加女性在STEM职业中的参与度。抗原是在生物体中引起免疫反应的分子。几种独特的抗原存在于某些人类癌细胞上。识别高亲和力结合这些抗原的T细胞受体（TCRs）是T细胞治疗癌症发展的关键瓶颈。该项目的目的是开发微流控技术，简化TCR筛选。这项技术将用于分选对抗原结合表现出生理反应的CD8+ T细胞。转录组测序将用于确定抗原识别TCR的最佳结构。这些序列的数据将用于设计具有最大特异性和选择性的T细胞，以识别癌细胞抗原。此外，对抗原具有更高亲和力的T细胞将被扩增，从而产生具有最佳抗肿瘤潜力的细胞群。提出的技术将用于确定引物和扩增的嵌合抗原受体（CAR） T细胞反应的程度和速度，这将使研究人员能够用特定的药物在期望的方向上“调整”细胞行为。最终目标是促进对免疫反应的理解，改善免疫疗法的生物制造和有效性。该奖项反映了美国国家科学基金会的法定使命，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。

Immune cells could form the basis of effective therapies for a variety of diseases. Producing these cells with uniform activity is difficult. This project will focus on developing a microfluidic device capable of sorting effective cells from ineffective ones. The effective population will be characterized. It is expected that the insight obtained will be useful in designing more effective chimeric antigen receptor (CAR) T cells. Interactive lectures and hands-on activities directed to high school students will be key components of the outreach activities supported by this project. Working through the Diversity and Women in Engineering Program to spark high school girls' interest in the future study of biotechnology and bioengineering should increase engagement of women in STEM careers. Antigens are molecules that induce an immune response in an organism. Several unique antigens are present on certain human cancer cells. Identifying T cell receptors (TCRs) that bind these antigens with high affinity is a key bottleneck in the development of T-cell therapy to treat cancer. The objective of this project is to develop microfluidic technology that simplifies TCR screening. This technology will be used to sort CD8+ T cells that display a physiological response to antigen binding. Transcriptomic sequencing will be used to identify the optimal structure of a TCR for antigen recognition. The data from these sequences will be used to engineer T cells with maximized specificity and selectivity for cancer cell antigen recognition. In addition, the sorted T cells, with a higher affinity for antigen, will be expanded to produce a cell population with optimal anti-tumor potential. The proposed technology will be used to determine the degree and rapidity of primed and expanded chimeric antigen receptor (CAR) T cell responses which will then enable researchers to “tune” cell behavior in a desired direction with specific drugs. The ultimate goal is to advance the understanding of immune responses and improve the biomanufacturing and effectiveness of immunotherapies.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.