Extended Data Fig. 4: NAD metabolism is altered in aged CD8+ T cells.
From: Age-associated nicotinamide adenine dinucleotide decline drives CAR-T cell failure
(a) Levels of CD38 within Tn, Tcm and Tem populations in young and old CD8+ T cells isolated from splenocytes (n = 5 biologically independent samples). (b) Proportion of Tcm cells on day 7 upon activation of naïve CD8+ T cells derived from young mice (8 weeks old) transduced with a CD38 OE vector and expanded under effector (IL-2) or memory (IL-7 IL-15) polarizing conditions (n = 9 biologically independent samples). (c) mtDNA/nDNA ratio in young CD8 + T cells overexpressing CD38 after 7 days of expansion with IL-7 and IL-15 (n = 9 biologically independent samples). (d) DNA damage, as assessed by pH2AX staining, in freshly isolated CD8+ T cells from young and old mice (n = 4 biologically independent samples). (e) pH2AX levels in CD38hi and CD38low CD8+ T cells from young and old mice (n = 4 biologically independent samples). HER2-directed CAR-T cells were generated from old mice and expanded in the presence of IL-7 and IL-15 ± NMN ± 78c ± Olaparib (PARPi). On day 7, NAD/NADH ratio (f) and SRC as assessed by Seahorse XFe96 Analysis (g) were determined (n = 3, in (G) dots represent technical replicates from one representative experiment). Data are represented as mean values +/- SEM. Statistical analysis was performed using unpaired t-test (A and D), paired t-test (B and C), two-way ANOVA (E), or one-way ANOVA with multiple comparisons (F and G), as appropiate. ns = p > 0.05.
