024 Asymmetric cell division for fate induction of chimeric antigen receptor (CAR) T cells

Early expansion and long-term persistence predict efficacy of genetically-engineered T cells. While this is thought to reflect induction effector memory cell populations provide both short-term clearance long-lasting remission, the cellular mechanisms fate after activation through synthetic receptors are unknown. A better understanding such processes could improve therapeutic outcome. Here we show that human cells engineered express chimeric antigen (CARs) undergo asymmetric division (ACD) with distinct proximal distal daughter adopt phenotypes, respectively. Using molecular proximity labeling distinguish first cells, demonstrate target-engaged CAR molecules remain on establish asymmetry between in proliferative pace, cytolytic function metabolic program. The single transcriptional program driven by c-myc, mTORC1 JAK-STAT3 (each adjusted p<0.001) resulting predominance glycolytic metabolism, features consistent differentiation. Conversely, utilize BACH-2, ETS-2 KLF2 shape their transcriptome rely oxidative phosphorylation, indicating a precursor phenotype. In vivo characterization two xenograft leukemia mouse models evaluate confirms superior elimination (n=14, p=0.0004) (n=12, p=0.0051) establishing these as precursors responsible for Collectively, studies uncover ACD novel framework differentiation influencing outcomes.