Fig. 3: BACH2^OE enhances function of 22/28-S.

a, Schematic of engineering approach using two lentiviral vectors with distinct transduction markers. b, Expression of transgenic BACH2. c, PD1 expression at the end of CAR T cell manufacture. d, Memory subset composition of CAR products at the end of manufacture. Subsets defined as naive cells (T_N cells, CD62L⁺CD45RO⁻), T_CM cells (CD62L⁺CD45RO⁺), T_EM cells (CD62L⁻CD45RO⁺) or terminal effector (T_Eff, CD62L⁻CD45RO⁻) cells. c,d, Representing n = 4 donors. e, Normalized expression of lineage-defining proteins quantified by mass cytometry. f, Principal component analysis representing variance in gene expression of CAR T cells at the end of manufacturing (n = 1 donor performed in technical triplicate). g, Heatmap of DEGs across 22/BB-S, 22/28-S and 22/28-S⁺BACH2^OE. h, Lineage signatures of genes upregulated in 22/28-S⁺BACH2^OE compared to 22/28-S as defined in the ImmuneSig database. i, Gene set variation analysis (GSVA) of 22/BB-S, 22/28-S and 22/28-S⁺BACH2^OE using naive, effector and exhausted signatures. j, Survival of Nalm6 over time in co-cultures with CAR T cells. k, Fold CAR T cell expansion on day 6 of co-culture with Nalm6. l, CAR T cell PD1 expression on day 6 of co-culture with Nalm6. m, Representative flow cytometry plots of co-expression of Eomes and TCF1 on day 3 of co-culture. For j–m, data represent n = 4 donors. n, BACH2 protein expression after CRISPR-mediated BACH2 disruption in CAR T cells. o, Memory subset composition of CAR T cell products at the end of manufacture. p, Nalm6 survival over time in co-culture with CAR T cells. q, Fold CAR T cell expansion on day 6 of co-culture with Nalm6. For n–q, representative data are from n = 2 donors. All data are presented as either individual values or mean values ± s.e.m. P < 0.05, P < 0.01, P < 0.001, P < 0.0001 by two-way ANOVA with Bonferroni’s adjustment for multiple comparisons. PC, principal component. Schematic in a created with BioRender.com.