Abstract
Chimeric antigen receptor (CAR) T cell therapy has proven to be a successful treatment option for leukemias and lymphomas. These encouraging outcomes underscore the potential of adoptive cell therapy for other oncology applications, namely, solid tumors. However, CAR T cells are yet to succeed in treating solid tumors. Unlike liquid tumors, solid tumors create a hostile tumor microenvironment (TME). CAR T cells must traffic to the TME, survive, and retain their function to eradicate the tumor. Nevertheless, there is no universal preclinical model to systematically test candidate CARs and CAR targets for their capacity to infiltrate and eliminate human solid tumors in vivo. Here, we provide a detailed protocol to evaluate human CAR CD4+ helper T cells and CD8+ cytotoxic T cells in immunodeficient (NSG) mice bearing antigen-expressing human solid tumors.
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Acknowledgements
This work was funded by Human Islet Research Network (HIRN) Emerging Leader in Type 1 Diabetes grant U24DK104162-07 (LMRF), American Cancer Society Institutional Research Grant IRG-19-137-20 (LMRF), and Cellular, Biochemical and Molecular Sciences Training Grant 5T32GM132055 (RWC). This publication was supported by the South Carolina Clinical & Translational Research Institute with an academic home at the Medical University of South Carolina CTSA NIH/NCATS grant number UL1 TR001450 (LMRF). The contents are solely the responsibility of the authors and do not necessarily represent the official views of the NIH or NCATS. Supported in part by the Flow Cytometry and Cell Sorting Shared Resource, Hollings Cancer Center, Medical University of South Carolina (P30 CA138313). pSLCAR-CD19-28z was a gift from Scott McComb (Addgene plasmid #135991). Figures 1 and 2 were created with BioRender.com. Statistical analyses in Fig. 3 were performed using GraphPad Prism for Mac. The flow cytometry results in Figs. 4, 5, and 6 were analyzed using FlowJo v10.8 Software (BD Life Sciences).
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Cochrane, R.W., Fiorentino, A., Allen, E., Robino, R.A., Quiroga, J., Ferreira, L.M.R. (2024). How to Test Human CAR T Cells in Solid Tumors, the Next Frontier of CAR T Cell Therapy. In: Siciliano, V., Ceroni, F. (eds) Cancer Immunotherapy. Methods in Molecular Biology, vol 2748. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3593-3_16
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DOI: https://doi.org/10.1007/978-1-0716-3593-3_16
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