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. 2024 May 14;57(5):1105-1123.e8.
doi: 10.1016/j.immuni.2024.04.006. Epub 2024 May 3.

Glucose-driven histone lactylation promotes the immunosuppressive activity of monocyte-derived macrophages in glioblastoma

Alessandra De Leo  1 , Alessio Ugolini  2 , Xiaoqing Yu  3 , Fabio Scirocchi  4 , Delia Scocozza  5 , Barbara Peixoto  1 , Angelica Pace  6 , Luca D'Angelo  7 , James K C Liu  8 , Arnold B Etame  8 , Aurelia Rughetti  6 , Marianna Nuti  6 , Antonio Santoro  7 , Michael A Vogelbaum  8 , Jose R Conejo-Garcia  9 , Paulo C Rodriguez  5 , Filippo Veglia  10
Affiliations

Glucose-driven histone lactylation promotes the immunosuppressive activity of monocyte-derived macrophages in glioblastoma

Alessandra De Leo et al. Immunity. .

Abstract

Immunosuppressive macrophages restrict anti-cancer immunity in glioblastoma (GBM). Here, we studied the contribution of microglia (MGs) and monocyte-derived macrophages (MDMs) to immunosuppression and mechanisms underlying their regulatory function. MDMs outnumbered MGs at late tumor stages and suppressed T cell activity. Molecular and functional analysis identified a population of glycolytic MDM expressing GLUT1 with potent immunosuppressive activity. GBM-derived factors promoted high glycolysis, lactate, and interleukin-10 (IL-10) production in MDMs. Inhibition of glycolysis or lactate production in MDMs impaired IL-10 expression and T cell suppression. Mechanistically, intracellular lactate-driven histone lactylation promoted IL-10 expression, which was required to suppress T cell activity. GLUT1 expression on MDMs was induced downstream of tumor-derived factors that activated the PERK-ATF4 axis. PERK deletion in MDM abrogated histone lactylation, led to the accumulation of intratumoral T cells and tumor growth delay, and, in combination with immunotherapy, blocked GBM progression. Thus, PERK-driven glucose metabolism promotes MDM immunosuppressive activity via histone lactylation.

Keywords: ER stress; PERK; brain cancer; glioblastoma; glycolysis; histone lactylation; immunosuppression; metabolism; myeloid cells; tumor-associated macrophages.

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Conflict of interest statement

Declaration of interests The authors declare no competing interests.

Figures

Figure 1.
Figure 1.. MDM replace MG during GBM progression and display potent immunosuppressive activity
(A) Gate strategy for TAMs in mouse brain tumors. (B) TAM proportion in mouse brain tumors (CT2A=5 mice, SB28=4 mice, GL261=5 mice) and in normal brain (Sham=5 mice). (C) TAM proportion at D0=5 mice, D12=4 mice, D24=5 mice, by FACS. (D) Suppression of T cell proliferation by TAMs from SB28 tumors, by FACS. (E) Kaplan–Meier survival curves (n = 17 isotype; n=25 αCD49d). (F) Gate strategy and TAM frequency in human GBM tumors (ND n=22; Rec n=14). (G) Proportion of TAMs, by FACS. (H) Survival analysis in GBM patients. (I) Suppression of T cell proliferation by TAMs from human GBM, by FACS.
Figure 2.
Figure 2.. Glycolytic profile of MDM and suppressive functions of GLUT1+ MDM.
(A) GSEA in MDM vs MG from mouse brain tumors (GL261). (B) Metabolite enrichment pathway analysis in MDM vs MG from mouse brain tumors (SB28). (C) ECAR in MDM and MG cells by Seahorse. (D) 2-NDGB uptake by FACS. (E) Proportion of 2-NDGBhigh cells in MDM from mouse brain tumors (SB28). (F) GLUT1 expression by FACS. (G) Proportion of GLUT1+ cells in MDM from mouse brain tumors, by FACS. (H) Glycolysis and Glycolytic Capacity by Seahorse. (I) Suppression of T cell proliferation by GLUT1− and GLUT1+ MDM from mouse brain tumors, FACS. (J) Suppression of T cell proliferation by Slc2a1fl/flCre+ MDM from mouse brain tumors, by FACS. (K) GLUT1 expression in human GBM tumors, by FACS. (L) Proportion of GLUT1+ cells in MDM from human GBM, by FACS. (M) Proportion of 2-NDGBhigh cells in MDM from human GBM, by FACS. (N) Suppression of T proliferation by human GLUT1− and GLUT1+MDM, by FACS.
Figure 3.
Figure 3.. Brain tumor-derived factors support high glycolysis and suppressive functions in BMDM.
(A) Uptake of 2-NDGB and GLUT1 expression in TES-BMDM, by FACS. (B) Basal OCR and ECAR analysis in TES-BMDM by Seahorse. (C) Basal ECAR, glycolysis and glycolytic capacity in TES-BMDM by Seahorse. (D) Suppression of T proliferation by TES-BMDM, by FACS. (E) Suppression of T proliferation by TES-BMDM treated with 1mM 2DG for 24h, by FACS. (F) Intracellular lactate levels in TES-BMDM treated with 2DG or with GNE-140 for 24h. (G) Intracellular lactate levels in TES-BMDM treated for the indicated times. (H) Suppression of T proliferation by TES-BMDM treated with GNE-140, by FACS. (I) Analysis of pHrodo Red in TAMs pulsed with lactic acid, by FACS. (J) Intracellular lactate levels in TAMs from brain tumors, by lactate colorimetric assay.
Figure 4.
Figure 4.. Glucose supports the expression of immunosuppressive genes in MDM.
(A) Heatmap of ATAC-seq of TAMs from mouse brain tumors (SB28 model), n=3. (B) Significant overlap between RNA-seq and ATAC-seq regulated genes in MDM vs MG. Up-regulated genes (RNA: padj<0.1005, log2FC>1; ATAC: padj<0.001, log2FC>1), down-regulated genes (RNA: padj<0.1005, log2FC< −1; ATAC: padj<0.001, log2FC< −1). (C) Il10 expression by RT-qPCR; n=10–13. (D) IL10 expression by WB. (E) IL10 expression by WB. (F) Il10 expression by RT-qPCR and IL10 release by ELISA, in TES-BMDM. (G) Kaplan–Meier survival curves (SB28). (H) Suppression of T proliferation by GLUT1+MDM from brain tumors, in presence of αIL10, by FACS. (I) Suppression of T proliferation by TES-BMDM in presence of αIL10, by FACS. (J) IL10 expression in MDM and MG from GBM patients, by RT-qPCR. (K) IL10 expression in TAMs from human GBM, by WB. (L) IL-10 expression in human TES-MDM by RT-qPCR; n=2. (M) Correlation analysis in GBM patients. (N) Survival analysis of GBM patients. (O) Il10 expression by TES-BMDM treated with 1mM 2DG, by RT-qPCR. (P) mIL10 release by ELISA, in TES-BMDM. (Q) Il10 expression in TES-BMDM treated with 5μM GNE-140 by RT-qPCR. (R) mIL10 release by TES-BMDM treated with GNE-140, by ELISA.
Figure 5.
Figure 5.. Glycolysis supports Kla in TES-BMDM.
(A) PanKla, PanKac expression in TES-BMDM, by WB. (B) PanKla, PanKac expression in TES-BMDM, by WB. (C) 13C incorporation in Kla histones in TES-BMDM loaded with U-13C6-glucose. (D) 13C incorporation in Kla histones in TES-BMDM loaded with U-13C6-glucose or 13C3-lactate. (E) PanKla expression in TAMs from mouse brain tumors, by WB. (F) PanKla and IL10 expression in MG and MDM from human brain tumors. (G) PanKla expression in GLUT1+MDM, by FACS. (H) PanKla expression in mouse GLUT1+ MDM, by WB. (I) PanKla and IL10 expression in mouse 2-NDGBhigh MDM, WB. (J) Kla marks at Il10 promoter in GLUT1+ MDM by CUT&RUN-qPCR. (K) Kla marks at Il10 promoter in mouse TES-BMDM, by CUT&RUN-qPCR. (L) PanKla, PanKac expression in TES-BMDM treated with 50nM CPI-1612 for 24h, by WB. (M). Il10 expression in TES-BMDM treated with CPI-1612, by RT-qPCR. (N) mIL10 release by TES-BMDM, by ELISA. (O) PanKla marks at Il10 promoters in TES-BMDM treated with CPI-1612, by CUT&RUN-qPCR.
Figure 6.
Figure 6.. PERK supports GLUT1 expression in immunosuppressive MDM in GBM tumors.
(A) PERK, pPERK expression in GLUT1+MDM from mouse brain tumors, by WB. (B) ATF4 in GLUT1+MDM, by WB. (C) Scl2a1 expression in MDM from Eif2ak3fl/flCre+ mice by RT-qPCR. (D) Proportion of GLUT1+ cells in MDM from Eif2ak3fl/flCre+ mice, by FACS. (E) Glycolysis and Glycolytic Capacity analysis in MDM from Eif2ak3fl/flCre+ mice, by Seahorse. (F) Suppression of T cell proliferation by MDM from Eif2ak3fl/flCre+, by FACS. (G) ATF4 at Slc2a1 promoter in THG-BMDM, by CUT&RUN-qPCR. (H) Scl2a1 expression in THG-BMDM by RT-qPCR. (I) 2-NDGB uptake by THG-BMDM by FACS. (J) Suppression of T cell proliferation by THG-BMDM, by FACS. (K) ATF4 expression in TES-BMDM, by WB. (L-M) Glycolysis analysis in TES-BMDM cells or THG-BMDM generated from Eif2ak3fl/flCre+ mice, by Seahorse. (N) PanKla expression in TES-BMDM generated from Eif2ak3fl/flCre+ mice, WB. (O) PanKla, panKac expression in THG-BMDM, by WB. (P) Kla marks at Il10 promoter in THG-BMDM, by CUT&RUN-qPCR.
Figure 7.
Figure 7.. PERK-deletion blocks tumor progression in combination with immunotherapy.
(A) Kaplan–Meier survival curves. (B) Kaplan–Meier survival curves of mice re-challenged with SB28, 60 days after brain tumor injection (SB28). (C) FACS-analysis of T cells at day14, after two doses of α4–1BB (day10, day12). (D) FACS-analysis of intratumoral TEFF, TEX, PEX collected at day14, after two doses of α4–1BB (day10, day12). (E) FACS-analysis of cytokine production by mouse intratumoral CD8 T cells at day14.
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References

    1. Ott M, Tomaszowski KH, Marisetty A, Kong LY, Wei J, Duna M, Blumberg K, Ji X, Jacobs C, Fuller GN, et al. (2020). Profiling of patients with glioma reveals the dominant immunosuppressive axis is refractory to immune function restoration. JCI Insight 5. 10.1172/jci.insight.134386. - DOI - PMC - PubMed
    1. Abdelfattah N, Kumar P, Wang C, Leu JS, Flynn WF, Gao R, Baskin DS, Pichumani K, Ijare OB, Wood SL, et al. (2022). Single-cell analysis of human glioma and immune cells identifies S100A4 as an immunotherapy target. Nat Commun 13, 767. 10.1038/s41467-022-28372-y. - DOI - PMC - PubMed
    1. Goswami S, Walle T, Cornish AE, Basu S, Anandhan S, Fernandez I, Vence L, Blando J, Zhao H, Yadav SS, et al. (2020). Immune profiling of human tumors identifies CD73 as a combinatorial target in glioblastoma. Nat Med 26, 39–46. 10.1038/s41591-019-0694-x. - DOI - PMC - PubMed
    1. Medikonda R, Dunn G, Rahman M, Fecci P, and Lim M (2021). A review of glioblastoma immunotherapy. J Neurooncol 151, 41–53. 10.1007/s11060-020-03448-1. - DOI - PubMed
    1. Sampson JH, Gunn MD, Fecci PE, and Ashley DM (2020). Brain immunology and immunotherapy in brain tumours. Nat Rev Cancer 20, 12–25. - PMC - PubMed

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