Generation of glucocorticoid resistant SARS-CoV-2 T-cells for adoptive cell therapy

Basar, R.; Uprety, N.; Ensley, E.; Daher, M.; Klein, K.; Martinez, F.; Aung, F.; Shanley, M.; Hu, B.; Gokdemir, E.; Mendt, M.; Reyes Silva, F.; Acharya, S.; Laskowski, T.; Muniz-Feliciano, L.; Banerjee, P.; Li, Y.; Li, S.; Melo Garcia, L.; Lin, P.; Shaim, H.; Yates, S. G.; Marin, D.; Kaur, I.; Rao, S.; Mak, D.; Lin, A.; Miao, Q.; Dou, J.; Chen, K.; Champlin, R.; Shpall, E. J.; Rezvani, K.

doi:10.1101/2020.09.15.298547

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bioRxiv · 10.1101/2020.09.15.298547

Generation of glucocorticoid resistant SARS-CoV-2 T-cells for adoptive cell therapy

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Abstract

Adoptive cell therapy with viral-specific T cells has been successfully used to treat life-threatening viral infections, supporting the application of this approach against COVID-19. We expanded SARS-CoV-2 T-cells from the peripheral blood of COVID-19-recovered donors and non-exposed controls using different culture conditions. We observed that the choice of cytokines modulates the expansion, phenotype and hierarchy of antigenic recognition by SARS-CoV-2 T-cells. Culture with IL-2/4/7 but not other cytokine-driven conditions resulted in >1000 fold expansion in SARS-CoV-2 T-cells with a retained phenotype, function and hierarchy of antigenic recognition when compared to baseline (pre-expansion) samples. Expanded CTLs were directed against structural SARS-CoV-2 proteins, including the receptor-binding domain of Spike. SARS-CoV-2 T-cells could not be efficiently expanded from the peripheral blood of non-exposed controls. Since corticosteroids are used for the management of severe COVID-19, we developed an efficient strategy to inactivate the glucocorticoid receptor gene (NR3C1) in SARS-CoV-2 CTLs using CRISPR-Cas9 gene editing.

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Basar, R., Uprety, N., Ensley, E., Daher, M., Klein, K., Martinez, F., Aung, F., Shanley, M., Hu, B., Gokdemir, E., Mendt, M., Reyes Silva, F., Acharya, S., Laskowski, T., Muniz-Feliciano, L., Banerjee, P., Li, Y., Li, S., Melo Garcia, L., Lin, P., Shaim, H., Yates, S. G., Marin, D., Kaur, I., Rao, S., Mak, D., Lin, A., Miao, Q., Dou, J., Chen, K., Champlin, R., Shpall, E. J., Rezvani, K.. 2020-09-15. Generation of glucocorticoid resistant SARS-CoV-2 T-cells for adoptive cell therapy. https://doi.org/10.1101/2020.09.15.298547

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