CAR +and CAR −T cells differentiate into an NK-like subset that is associated with increased inflammatory cytokines following infusion

Abstract Chimeric antigen receptor (CAR) T cells have demonstrable efficacy in treating B-cell malignancies. Factors such as product composition, lymphodepletion and immune reconstitution are known to influence CAR +T cell survival persistence, however the determinants of cells’ fate differentiation how patients’ reconstituting system therapy outcomes remain poorly understood. We applied single multi-omics analysis RNA, protein clonal profiles PBMC patients receiving donor-derived products from a piggybac CAR19 clinical trial (N=8 patients). used these data reconstruct trajectory, which explained observed phenotype +and −T within first 30 days post-infusion. Following initial lympho-depletion, endogenous −CD8 +, γδ CAR+ clonally expanded, differentiated across heterogenous phenotypes. They transitioned dominant resting or proliferating state into precursor exhausted cells, finally terminal NK-like phenotype. The subset was associated with increased serum concentrations proinflammatory cytokines including IL-12 IL-18. Validation on published cohort (61 patients) showed that this outcome at 6 months. Finally, we analysed detail one patient developed induced lymphoma discussed impact early phase determining unexpected outcome. These results demonstrate CAR- share profile may be determined by non-CAR derived signals immune-recovery National Health And Medical Research Funding (Australia) (APP1121643, APP1128416) BD Bioscience