Label-free in vitro assays predict the potency of anti-disialoganglioside chimeric antigen receptor T-cell products |
| |
| Affiliation: | 1. Regenerative Bioscience Center, University of Georgia, Athens, Georgia, USA;2. Division of Neuroscience, Biomedical and Health Sciences Institute, University of Georgia, Athens, Georgia, USA;3. Edgar L. Rhodes Center for Animal and Dairy Science, College of Agriculture and Environmental Science, University of Georgia, Athens, Georgia, USA;4. Department of Biochemistry & Molecular Biology, Complex Carbohydrate Research Center, University of Georgia, Athens, Georgia, USA;5. Institute of Bioinformatics, University of Georgia, Athens, Georgia, USA;6. Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, Wisconsin USA;7. Axion BioSystems, Atlanta, Georgia USA;8. Department of Pediatrics, University of Wisconsin School of Medicine and Public Health, Madison, Wisconsin USA;9. University of Wisconsin Carbone Cancer Center, University of Wisconsin-Madison, Madison, Wisconsin USA;10. Department of Pathology, Northwestern University Feinberg School of Medicine, Chicago, Illinois USA;11. Evolved Analytics LLC, Rancho Santa Fe, California, USA;12. Morgridge Institute for Research, Madison, Wisconsin, USA |
| |
| Abstract: | Background aimsChimeric antigen receptor (CAR) T cells have demonstrated remarkable efficacy against hematological malignancies; however, they have not experienced the same success against solid tumors such as glioblastoma (GBM). There is a growing need for high-throughput functional screening platforms to measure CAR T-cell potency against solid tumor cells.MethodsWe used real-time, label-free cellular impedance sensing to evaluate the potency of anti-disialoganglioside (GD2) targeting CAR T-cell products against GD2+ patient-derived GBM stem cells over a period of 2 days and 7 days in vitro. We compared CAR T products using two different modes of gene transfer: retroviral transduction and virus-free CRISPR-editing. Endpoint flow cytometry, cytokine analysis and metabolomics data were acquired and integrated to create a predictive model of CAR T-cell potency.ResultsResults indicated faster cytolysis by virus-free CRISPR-edited CAR T cells compared with retrovirally transduced CAR T cells, accompanied by increased inflammatory cytokine release, CD8+ CAR T-cell presence in co-culture conditions and CAR T-cell infiltration into three-dimensional GBM spheroids. Computational modeling identified increased tumor necrosis factor α concentrations with decreased glutamine, lactate and formate as being most predictive of short-term (2 days) and long-term (7 days) CAR T cell potency against GBM stem cells.ConclusionsThese studies establish impedance sensing as a high-throughput, label-free assay for preclinical potency testing of CAR T cells against solid tumors. |
| |
| Keywords: | |
| 本文献已被 ScienceDirect 等数据库收录！ |
|
