Fate induction in CD8 chimeric antigen receptor T-cells through asymmetric cell division

Early expansion and long-term persistence predict efficacy of chimeric antigen receptor T-cells (CARTs), but mechanisms governing effector versus memory CART differentiation and whether asymmetric cell division (ACD) induces differential fates in human CARTs remain unclear. We use target-induced proximity labeling to isolate first-division proximal--daughter and distal-daughter CARTs to interrogate TCR clonality, surface proteome, and transcriptome at the single-cell level. First-division proximal (LIPSTIC label positive, first cell division by CTV stain), first-division distal (LIPSTIC label negative, first cell division), activated-undivided (LIPSTIC label positive, zero cell division), and resting (LIPSTIC label negative, zero cell division) CARTs were isolated using fluorescence-activated cell sorting and analyzed by single cell sequencing.