Novel co-culture strategies of tumor organoids with autologous T-cells reveal clinically relevant combinations of immune-checkpoint and targeted therapies

Podaza, E.; Capuano, J.; Al Assaad, M.; Kuo, H.-h.; Markowitz, G.; Irizarry, A.; Ravichandran, H.; Ackermann, S.; Kane, T.; Manohar, J.; Sigouros, M.; Moyer, J.; Bhinder, B.; Chandra, P.; Malbari, M.; Boehnke, K.; Mosquera, J. M.; Mittal, V.; Sboner, A.; Gokozan, H.; Altorki, N.; Elemento, O.; Martin, M. L.

doi:10.1101/2023.07.05.546622

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bioRxiv · 10.1101/2023.07.05.546622

Novel co-culture strategies of tumor organoids with autologous T-cells reveal clinically relevant combinations of immune-checkpoint and targeted therapies

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Abstract

Patient derived tumor organoids (PDTOs) have become relevant pre-clinical models for therapeutic modelling since they highly recapitulate patients response to treatment. Nevertheless, their value for immunotherapy modelling has not been fully explored. We developed a tumor processing protocol that enable the establishment of PDTOs and tumor infiltrating lymphocytes (TILs) isolation. By the optimization of functional assays, we compared the T-cells effector functions of matching PBMCs and TILs, demonstrating that PBMCs after co-culture and TILs after initial expansion display similar responses. In addition, the evaluation of cytokine production by fluorospot in combination with an image-based killing assay enable the screening of different immune-checkpoint inhibitors as well as its combination with target inhibitors. Our proof-of-concept functional assays showed the potential and versatility of PDTOs and T-cells co-culture systems for immunotherapy screening. The optimization of scalable functional assays downstream co-culture represents a significant step forward to increase the value of PDTOs as pre-clinical models for immunotherapeutic screens.

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Podaza, E., Capuano, J., Al Assaad, M., Kuo, H.-h., Markowitz, G., Irizarry, A., Ravichandran, H., Ackermann, S., Kane, T., Manohar, J., Sigouros, M., Moyer, J., Bhinder, B., Chandra, P., Malbari, M., Boehnke, K., Mosquera, J. M., Mittal, V., Sboner, A., Gokozan, H., Altorki, N., Elemento, O., Martin, M. L.. 2023-07-05. Novel co-culture strategies of tumor organoids with autologous T-cells reveal clinically relevant combinations of immune-checkpoint and targeted therapies. https://doi.org/10.1101/2023.07.05.546622

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