Inhibition of 2-hydroxyglutarate elicits metabolic reprogramming and mutant IDH1 glioma immunity in mice
Padma Kadiyala, … , Pedro R. Lowenstein, Maria G. Castro
Padma Kadiyala, … , Pedro R. Lowenstein, Maria G. Castro
Published December 17, 2020
Citation Information: J Clin Invest. 2021;131(4):e139542. https://doi.org/10.1172/JCI139542.
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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 10

D-2HG does not inhibit T cell proliferation and activation.

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D-2HG does not inhibit T cell proliferation and activation. (A) Experime...
(A) Experimental design to analyze the impact of D-2HG on T cell proliferation. (B) Flow plots show representative CFSE stains of unstimulated splenocytes (inactivated T cells), splenocytes undergoing proliferation in response to 100 nM SIINFEKL (activated T cells), and the effect of 0.25 mM D-2HG on SIINFEKL-induced T cell proliferation. (C) Quantification of OT-1 splenocytes undergoing T cell proliferation. ***P < 0.001, 1-way ANOVA. Bars represent mean ± SEM (n = 3 technical replicates). (D) Quantification of IFN-γ levels in the supernatants of OT-1 splenocytes stimulated with SIINFEKL in the presence of D-2HG. IFN-γ levels were assessed by ELISA. (E) Representative flow plots of OT-1 splenocytes incubated with 0.25 mM D-2HG in the presence of 100 nM SIINFEKL peptide for 4 days and stained with Annexin V-FITC and propidium iodide (PI). Live activated T cells (CD3+/CD8+/IFN-γ+) were identified as Annexin V negative and PI negative. Dead cells undergoing early apoptosis were identified as Annexin V positive and PI negative. Dead T cells undergoing late apoptosis were identified as Annexin V positive and PI positive. Bars represent quantitative analysis of the distribution of live and dead cells during 4-day incubation period with D-2HG (n = 3 technical replicates).