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

Basar, Rafet; Uprety, Nadima; Ensley, Emily; Daher, May; Klein, Kimberly; Martinez, Fernando; Aung, Fleur; Shanley, Mayra; Hu, Bingqian; Gokdemir, Elif; Nunez Cortes, Ana Karen; Mendt, Mayela; Silva, Francia Reyes; Acharya, Sunil; Laskowski, Tamara; Muniz-Feliciano, Luis; Banerjee, Pinaki P.; Li, Ye; Li, Sufang; Garcia, Luciana Melo; Lin, Paul; Shaim, Hila; Yates, Sean G.; Marin, David; Kaur, Indreshpal; Rao, Sheetal; Mak, Duncan; Lin, Angelique; Miao, Qi; Dou, Jinzhuang; Chen, Ken; Champlin, Richard E.; Shpall, Elizabeth J.; Rezvani, Katayoun

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 expand SARS-CoV-2 T-cells from the peripheral blood of COVID-19-recovered donors and non-exposed controls using different culture conditions. We observe 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 result 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 are directed against structural SARS-CoV-2 proteins, including the receptor-binding domain of Spike. SARS-CoV-2 T-cells cannot be efficiently expanded from the peripheral blood of non-exposed controls. Since corticosteroids are used for the management of severe COVID-19, we propose an efficient strategy to inactivate the glucocorticoid receptor gene (NR3C1) in SARS-CoV-2 CTLs using CRISPR-Cas9 gene editing.