Extended Data Fig. 7: Inhibition of IDH2 induces memory features in human T cells.
From: Reductive carboxylation epigenetically instructs T cell differentiation
a,b, MFI of CD62L (a) and representative histogram (b) at day 9 post-activation on the surface of human CD8+ T cells isolated from healthy volunteer PBMCs, measured by flow cytometry. PBMCs from healthy donors were activated with anti-CD3/CD28 beads for 5 days and cultured 4 additional days prior phenotypic analyses. The cells were cultured in the presence of DMSO or IDH2i during the entire experiment (a–c). (n = 7 donors, pooled data from 2 independent experiments). c, Percentages of human CD8+ TSCM-like cells, characterized as CD45RA+, CD62L+, CD45RO−, CCR7+, CD95+, CD28+, CD27+. (n = 7 donors, pooled data from 2 independent experiments). d, Experimental set-up of NAML6 tumour experiment. Human T cells were transduced with anti-CD19 CAR T cells, expanded during 12 days in the presence of DMSO or IDH2i and subsequently transferred into tumour-bearing NSG mice. e–g, Human MILs from patients with multiple myeloma were activated with anti-CD3/CD28 beads for 5 days and cultured 4 additional days prior phenotypic analyses. The cells were cultured in the presence of DMSO or IDH2i during the entire experiment. Shown here is MFI of CD62L (e) and representative histogram (f), and percentage of TSCM (g) at day 9 post-activation in human CD8+ MILs. (n = 6 donors, pooled data from 2 independent experiments). h, Experimental set-up of NCI-H929 tumour experiment. Human T cells were transduced with anti-BCMA CAR T cells, expanded during 10 days in the presence of DMSO or IDH2i and subsequently transferred into tumour-bearing NSG mice. i, BLI quantification of total flux (p/s) (representative of 2 independent experiments). j–l, Fold-change expansion at day 9 post-activation of human PBMCs (j), human MILs (k) and human CAR T cells (l) cultured with DMSO or IDH2i. (n = 3 (j), n = 6 (k) and n = 2 donors (l), pooled data from 2 independent experiments (k). m, Representation (row z-scores) of differentially accessible regions in mouse CD8+ T cells after DMSO or IDH2i conditioning (ATAC-seq). n, Bar plots representing log2 fold change in mRNA expression of Sell, Tcf7 and Ccr7 upon IDH2i-compared to DMSO-conditioned CD8+ T cells (RNA-seq). Data represent mean ± s.e.m. and were analysed by paired two-tailed Student’s t-test (a,c,e,g,j–m).
