EAGER BIOMANUFACTURING: A Microscale Testbed to Assay and Manufacture CAR T-Cell Immunotherapies

University Of Wisconsin-Madison, Madison WI

Investigators

Abstract

1645123 - Saha Genetically-engineered human T-cells that express chimeric antigen receptors (CARs) have been used to target cancer associated antigens. While CAR T-cells have had success in treating hematological (blood related) malignancies, they are only just starting to be used to combat solid tumors. Currently, results in solid tumor trials have not been as promising as those seen in leukemia and have highly inconsistent outcomes. One possible cause of the differences in response may be due to the population heterogeneity of CAR activity among T-cells. Efficient biomanufacturing platforms are needed to define the functional heterogeneity after genetic engineering and to robustly generate large quantities of high quality CAR T-cells. Here, the investigators propose that the isolation of functionally cytotoxic CAR T-cell fractions is important for the efficacy of immunotherapies. Successful completion of this project will develop a flexible, cost-effective, and scalable microscale culture system to isolate functional gene-edited CAR T-cells, expand the fraction, and assess in vivo functionality. This project will develop a novel paradigm on how to approach gene-editing processes and heterogeneity in T-cell biomanufacturing. The investigators propose to fractionate complex modified T-cell cultures based on CAR functionality. The microscale approach will be non-destructive after gene editing and pre-patient infusion. Then, the investigators will provide one of the first analyses of CAR heterogeneity and fractionation. Defined CAR T-cell populations will be profiled in vivo in xenograft mouse models of solid cancer. In the long-term, a microscale testbed could be used to increase the level of cell-intrinsic complexity programmed into immunotherapies by synthetic biologists and the level of cell-extrinsic complexity built into functional assays by tissue engineers. Education and outreach activities will also train cell engineers at the graduate and undergraduate levels, and will provide outreach to K-12 students, K-12 teachers, and the general public on aspects of gene editing and biomanufacturing.