Figure 6. GLUT3-dependent histone acetylation promotes Th17 cell effector function.

(A) Effects of 2-Hydroxycitrate (2-HC), Soraphen A (SorA) or 5-Tetradecyloxy-2-furonic acid (TOFA) to inhibit ATP-citrate lyase (ACLY), acetyl-CoA carboxylases (ACC1/2) or fatty acid synthase (FAS), respectively. (B) Analysis of IL-17 expression in Th17 cells treated with 2-HC, SorA or TOFA during differentiation (72 h) or re-stimulation (5 h). (C-J) GLUT3-dependent acetyl-CoA controls the epigenetic re-programming of Th17 cells. (C and D) Analysis of global histone 3 (H3) acetylation at lysins K9/14 (C) and K27 (D) in WT and GLUT3-deficient Th17 cells; means ± SEM of 5=9 mice. (E) Glucose-derived incorporation of [¹⁴C] carbons into histones of WT, GLUT3- and ACLY-deficient Th17 cells; means ± SEM of 3 mice. (F) Analysis of IL-17 expression in GLUT3-deficient Th17 cells treated with 10 mM Panobinostat for 24 h; means ± SEM of 4-7 mice. (G-K) Genome-wide analysis of H3 K9/14 acetylation in WT and GLUT3-deficient Th17 cells by chromatin immunoprecipitation followed by DNA-sequencing (ChIP-seq). (G) Heatmaps using depth-normalized coverages of global K9/14 histone acetylation of WT and GLUT3-deficient Th17 cells relative to input samples. (H) Analysis of H3 K9/14 acetylation at the Il17a and Il17f gene cluster in WT and GLUT3-deficient Th17. Promoters (green shading), enhancers and other conserved noncoding regions (CNS, orange shading) were determined using ATAC-seq datasets of Th17 cells (Qiu et al., 2020). (I and J) Quantification of K9/14 (I) and K27 (J) acetylation in WT and GLUT3-deficient Th17 cells at the CNS-2, Il17a and Il17f promoter using ChIP-qPCR; n=5-6 mice. (K) Analysis of H3 K9/14 acetylation at the Il2 locus in WT and GLUT3-deficient Th17 cells. *, p<0.05; **, p<0.01, ***, p<0.001 by unpaired Student’s t-test (C-F), (I) and (J).