TY - JOUR
T1 - Three-Dimensional Modeling of Solid Tumors and Their Microenvironment to Evaluate T Cell Therapy Efficacy In Vitro
AU - Pscheid, Ronja
AU - Drent, Esther
AU - Wienke, Judith
AU - Strijker, Josephine
AU - Throsby, Mark
AU - Molenaar, Jan
N1 - Copyright © 2023 by The American Association of Immunologists, Inc.
PY - 2023/7/15
Y1 - 2023/7/15
N2 - Immunotherapy development for solid tumors remains challenging, partially due to a lack of reproducible, cost-effective in vitro three-dimensional (3D) models to mimic the heterogeneous and complex tumor microenvironment. Here, we investigate the cellular anti-tumor reactivity of αβ T cells engineered to express a defined γδ TCR (TEG A3). For that purpose, we developed a 3D cytotoxicity assay targeting cell line–derived spheroids or patient-derived tumor organoids formed in serum-free media. Tumor cell lysis by TEG A3 was monitored using the Incucyte S3 live-cell imaging system with the apoptosis marker caspase 3/7 green and endpoint readouts of IFN-γ secretion in the supernatant. The 3D cytotoxicity assay model system was able to adequately demonstrate TEG A3 reactivity toward targets expressing an isoform of CD277 (CD277J). To obtain a more complex heterogeneous tumor microenvironment, patient-derived organoids were mixed with unmatched patient-derived fibroblasts or matched cancer-associated fibroblasts. In all assays, we demonstrated the tumor target specificity of TEG A3, lysing tumor cells within 48 h. Our study demonstrates the utility of complex 3D cytotoxicity assay model systems incorporating the tumor microenvironment in the functional evaluation of T cell–based adoptive immunotherapy, providing a useful platform for early-stage preclinical development of immunotherapies.
AB - Immunotherapy development for solid tumors remains challenging, partially due to a lack of reproducible, cost-effective in vitro three-dimensional (3D) models to mimic the heterogeneous and complex tumor microenvironment. Here, we investigate the cellular anti-tumor reactivity of αβ T cells engineered to express a defined γδ TCR (TEG A3). For that purpose, we developed a 3D cytotoxicity assay targeting cell line–derived spheroids or patient-derived tumor organoids formed in serum-free media. Tumor cell lysis by TEG A3 was monitored using the Incucyte S3 live-cell imaging system with the apoptosis marker caspase 3/7 green and endpoint readouts of IFN-γ secretion in the supernatant. The 3D cytotoxicity assay model system was able to adequately demonstrate TEG A3 reactivity toward targets expressing an isoform of CD277 (CD277J). To obtain a more complex heterogeneous tumor microenvironment, patient-derived organoids were mixed with unmatched patient-derived fibroblasts or matched cancer-associated fibroblasts. In all assays, we demonstrated the tumor target specificity of TEG A3, lysing tumor cells within 48 h. Our study demonstrates the utility of complex 3D cytotoxicity assay model systems incorporating the tumor microenvironment in the functional evaluation of T cell–based adoptive immunotherapy, providing a useful platform for early-stage preclinical development of immunotherapies.
KW - Humans
KW - Neoplasms/therapy
KW - T-Lymphocytes
KW - Immunotherapy, Adoptive/methods
KW - Immunotherapy
KW - Cell- and Tissue-Based Therapy
KW - Tumor Microenvironment
UR - https://www.mendeley.com/catalogue/751b4bc8-4382-3b4c-97a9-84d4517e0292/
U2 - 10.4049/jimmunol.2200573
DO - 10.4049/jimmunol.2200573
M3 - Article
C2 - 37294309
SN - 0022-1767
VL - 211
SP - 229
EP - 240
JO - Journal of Immunology
JF - Journal of Immunology
IS - 2
ER -