Inhibition of 2-hydroxyglutarate elicits metabolic reprogramming and mutant IDH1 glioma immunity in mice
Padma Kadiyala, Stephen V. Carney, Jessica C. Gauss, Maria B. Garcia-Fabiani, Santiago Haase, Mahmoud S. Alghamri, Felipe J. Núñez, Yayuan Liu, Minzhi Yu, Ayman Taher, Fernando M. Nunez, Dan Li, Marta B. Edwards, Celina G. Kleer, Henry Appelman, Yilun Sun, Lili Zhao, James J. Moon, Anna Schwendeman, Pedro R. Lowenstein, Maria G. Castro
Padma Kadiyala, Stephen V. Carney, Jessica C. Gauss, Maria B. Garcia-Fabiani, Santiago Haase, Mahmoud S. Alghamri, Felipe J. Núñez, Yayuan Liu, Minzhi Yu, Ayman Taher, Fernando M. Nunez, Dan Li, Marta B. Edwards, Celina G. Kleer, Henry Appelman, Yilun Sun, Lili Zhao, James J. Moon, Anna Schwendeman, Pedro R. Lowenstein, Maria G. Castro
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Research Article Immunology Neuroscience

Inhibition of 2-hydroxyglutarate elicits metabolic reprogramming and mutant IDH1 glioma immunity in mice

Abstract

Mutant isocitrate dehydrogenase 1 (IDH1-R132H; mIDH1) is a hallmark of adult gliomas. Lower grade mIDH1 gliomas are classified into 2 molecular subgroups: 1p/19q codeletion/TERT-promoter mutations or inactivating mutations in α-thalassemia/mental retardation syndrome X-linked (ATRX) and TP53. This work focuses on glioma subtypes harboring mIDH1, TP53, and ATRX inactivation. IDH1-R132H is a gain-of-function mutation that converts α-ketoglutarate into 2-hydroxyglutarate (D-2HG). The role of D-2HG within the tumor microenvironment of mIDH1/mATRX/mTP53 gliomas remains unexplored. Inhibition of D-2HG, when used as monotherapy or in combination with radiation and temozolomide (IR/TMZ), led to increased median survival (MS) of mIDH1 glioma–bearing mice. Also, D-2HG inhibition elicited anti–mIDH1 glioma immunological memory. In response to D-2HG inhibition, PD-L1 expression levels on mIDH1-glioma cells increased to similar levels as observed in WT-IDH gliomas. Thus, we combined D-2HG inhibition/IR/TMZ with anti–PDL1 immune checkpoint blockade and observed complete tumor regression in 60% of mIDH1 glioma–bearing mice. This combination strategy reduced T cell exhaustion and favored the generation of memory CD8+ T cells. Our findings demonstrate that metabolic reprogramming elicits anti–mIDH1 glioma immunity, leading to increased MS and immunological memory. Our preclinical data support the testing of IDH-R132H inhibitors in combination with IR/TMZ and anti-PDL1 as targeted therapy for mIDH1/mATRX/mTP53 glioma patients.

Authors

Padma Kadiyala, Stephen V. Carney, Jessica C. Gauss, Maria B. Garcia-Fabiani, Santiago Haase, Mahmoud S. Alghamri, Felipe J. Núñez, Yayuan Liu, Minzhi Yu, Ayman Taher, Fernando M. Nunez, Dan Li, Marta B. Edwards, Celina G. Kleer, Henry Appelman, Yilun Sun, Lili Zhao, James J. Moon, Anna Schwendeman, Pedro R. Lowenstein, Maria G. Castro

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Figure 1

IDH1-R132H inhibition in mIDH1 mouse-NSs and human-GCs confers radiosensitivity and promotes release of DAMPs.

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IDH1-R132H inhibition in mIDH1 mouse-NSs and human-GCs confers radiosens...
(A) Mouse-NS cultures were generated from WT-IDH or mIDH1 glioma models. (B–E) Clonogenic assay of mIDH1 and WT-IDH mouse-NSs and human-GCs. (B and C) Mouse-NS were treated with 0–8 Gy ionizing radiation (IR) and 1.5 μM AGI-5198 (red) or DMSO vehicle (blue). (D and E) Human-GCs SJGBM2 (WT-IDH) and MGG119 (mIDH1) were treated with 0–20 Gy IR and 5 μM AGI-5198 (red) or DMSO vehicle (blue). ****P < 0.0001, nonlinear regression. Bars represent mean ± SEM (n = 3 technical replicates). Representative images of colonies stained with crystal violet in each treatment group are shown under the survival fraction graphs. (F–I) Calreticulin (CRT), ATP, and HMGB1 expression levels within mIDH1 mouse-NSs and human-GCs. Mouse-NS were treated with 3 Gy IR and 1.5 μM AGI-5198 for 72 hours. Human-GC were treated with 10 Gy IR and 5 μM AGI-5198 for 72 hours. Quantification of CRT expression on mIDH1 mouse-NSs and human-GCs after treatment is shown in F and G, respectively. Representative histograms display CRT marker’s expression levels (black, isotype control; green, DMSO vehicle; light blue, IR; red, AGI-5198; dark blue, AGI5198 + IR). ****P < 0.0001, **P < 0.001, 1-way ANOVA. Bars represent mean ± SEM (n = 3 technical replicates). (H) Quantification of ATP release in the supernatant of mIDH1 mouse-NSs and human-GCs. *P < 0.01; **P < 0.01; ***P < 0.001, 1-way ANOVA. Bars represent mean ± SEM (n = 3 technical replicates). (I) Quantification of HMGB1 release in the supernatant of mIDH1 mouse-NSs and human-GCs. **P < 0.01; ***P < 0.001; ****P < 0.0001, 1-way ANOVA. Bars represent mean ± SEM (n = 3 technical replicates).