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。生产具有均匀活性的这些细胞是困难的。该项目将专注于开发一种能够从无效细胞中分选有效细胞的微流体装置。将对有效群体进行表征。预计所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。针对高中生的互动讲座和实践活动将是该项目支持的外联活动的关键组成部分。通过多样性和妇女参与工程项目，激发高中女生对未来生物技术和生物工程研究的兴趣，应该会增加妇女在STEM职业中的参与。抗原是在生物体中诱导免疫应答的分子。某些人类癌细胞上存在几种独特的抗原。鉴定以高亲和力结合这些抗原的T细胞受体（TCR）是开发治疗癌症的T细胞疗法的关键瓶颈。该项目的目标是开发简化TCR筛选的微流控技术。该技术将用于分选对抗原结合表现出生理反应的CD8+ T细胞。转录组测序将用于鉴定用于抗原识别的TCR的最佳结构。来自这些序列的数据将用于工程化T细胞，使其对癌细胞抗原识别具有最大的特异性和选择性。此外，对抗原具有更高亲和力的分选的T细胞将被扩增以产生具有最佳抗肿瘤潜力的细胞群。 该技术将用于确定引发和扩增的嵌合抗原受体（CAR）T细胞反应的程度和速度，然后使研究人员能够用特定药物将细胞行为“调整”到所需的方向。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。

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.