γδ T cell-mediated cytotoxicity against patient-derived healthy and cancer cervical organoids

Dong, J.; Holthaus, D.; Peters, C.; Koster, S.; Ehsani, M.; Quevedo-Olmos, A.; Berger, H.; Zarobkiewicz, M.; Mangler, M.; Kumar Gurumurthy, R.; Hedemann, N.; Chumduri, C.; Kabelitz, D.; Meyer, T. F.

doi:10.1101/2023.07.25.550464

bioRxiv · 10.1101/2023.07.25.550464

γδ T cell-mediated cytotoxicity against patient-derived healthy and cancer cervical organoids

Dong, J.·Holthaus, D.·Peters, C.·Koster, S.·Ehsani, M.·Quevedo-Olmos, A.·Berger, H.·Zarobkiewicz, M.·Mangler, M.·Kumar Gurumurthy, R.·Hedemann, N.·Chumduri, C.·Kabelitz, D.·Meyer, T. F.

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Abstract

Cervical cancer is a leading cause of death among women globally, primarily driven by high-risk papillomaviruses. However, the effectiveness of chemotherapy is limited, underscoring the potential of personalized immunotherapies. Patient-derived organoids, which possess cellular heterogeneity, proper epithelial architecture and functionality, and long-term propagation capabilities offer a promising platform for developing viable strategies. In addition to {beta} T cells and natural killer (NK) cells, {gamma}{delta} T cells represent a cell population with significant therapeutic potential against both hematologic and solid tumours. To evaluate the efficacy of {gamma}{delta} T cells in cervical cancer treatment, we generated patient-derived healthy and cancer ectocervical organoids. Furthermore, we examined transformed healthy organoids, expressing HPV16 oncogenes E6 and E7. We analysed the effector function of in vitro expanded {gamma}{delta} T cells upon co-culture with organoids. Our findings demonstrated that healthy cervical organoids were less susceptible to {gamma}{delta} T cell-mediated cytotoxicity compared to HPV-transformed organoids and cancerous organoids. To identify the underlying pathways involved in this observed cytotoxicity, we performed bulk-RNA sequencing on the organoid lines, revealing differences in DNA-damage and cell cycle checkpoint pathways, as well as transcription of potential {gamma}{delta} T cell ligands. We validated these results using immunoblotting and flow cytometry. We also demonstrated the involvement of BTN3A1 and BTN2A1, crucial molecules for {gamma}{delta} T cell activation, as well as differential expression of PDL1/CD274 in cancer, E6/E7+ and healthy organoids. Interestingly, we observed a significant reduction in cytotoxicity upon blocking MSH2, a protein involved in DNA mismatch-repair. In summary, we established a co-culture system of {gamma}{delta} T cells with cervical cancer organoids, providing a novel in vitro model to optimize innovative patient-specific immunotherapies for cervical cancer.

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Dong, J., Holthaus, D., Peters, C., Koster, S., Ehsani, M., Quevedo-Olmos, A., Berger, H., Zarobkiewicz, M., Mangler, M., Kumar Gurumurthy, R., Hedemann, N., Chumduri, C., Kabelitz, D., Meyer, T. F.. 2023-07-26. γδ T cell-mediated cytotoxicity against patient-derived healthy and cancer cervical organoids. https://doi.org/10.1101/2023.07.25.550464

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