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

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.