Pyruvate kinase M2 is requisite for Th1 and Th17 differentiation
Michihito Kono, … , Nobuya Yoshida, George C. Tsokos
Michihito Kono, … , Nobuya Yoshida, George C. Tsokos
Published June 20, 2019
Citation Information: JCI Insight. 2019;4(12):e127395. https://doi.org/10.1172/jci.insight.127395.
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Research Article Immunology

Pyruvate kinase M2 is requisite for Th1 and Th17 differentiation

Abstract

Th1 and Th17 are important in the pathogenesis of autoimmune diseases and they depend on glycolysis as a source of energy. T cell antigen receptor signaling phosphorylates a serine/threonine kinase, calcium/calmodulin–dependent protein kinase IV (CaMK4), and promotes glycolysis. Based on these findings we hypothesized that CaMK4 promotes glycolysis. Camk4-deficient CD4+ T cells and cells treated with a CaMK4 inhibitor had less glycolysis compared with their counterparts. Pull-down of CaMK4 and mass spectrometry identified pyruvate kinase muscle isozyme (PKM), the final rate-limiting enzyme in glycolysis, as a binding partner. Coimmunoprecipitation and Western blotting showed that CaMK4 interacts directly with PKM2. Camk4-deficient CD4+ T cells displayed decreased pyruvate kinase activity. Silencing or pharmacological inhibition of PKM2 reduced glycolysis and in vitro differentiation to Th1 and Th17 cells, while PKM2 overexpression restored Th17 cell differentiation. Treatment with a PKM2 inhibitor ameliorated experimental autoimmune encephalomyelitis and CD4+ T cells treated with PKM2 inhibitor or Pkm2-shRNA caused limited disease activity in an adoptive cell transfer model of experimental autoimmune encephalomyelitis. Our data demonstrate that CaMK4 binds to PKM2 and promotes its activity, which is requisite for Th1 and Th17 differentiation in vitro and in vivo. PKM2 represents a therapeutic target for T cell–dependent autoimmune diseases.

Authors

Michihito Kono, Kayaho Maeda, Irina Stocton-Gavanescu, Wenliang Pan, Masataka Umeda, Eri Katsuyama, Catalina Burbano, Seo Yeon K. Orite, Milena Vukelic, Maria G. Tsokos, Nobuya Yoshida, George C. Tsokos

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Figure 6

Pkm2-shRNA–transfected Th17 cells reduce disease activity in an adoptive cell transfer EAE model.

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Pkm2-shRNA–transfected Th17 cells reduce disease activity in an adoptiv...
(A–C) Naive CD4+ T cells from 2D2 mice were cultured under Th17-polarizing conditions and transfected with Pkm2-shRNA or control shRNA. These cells were transferred into Rag1-deficient mice intravenously and EAE was induced. (A) Clinical scores of recipient mice are shown. Cumulative results of 2 independent experiments with 3–4 mice per group are shown (mean ± SEM); n = 7–8. (B) Cumulative results of body weight changes of recipient mice (mean ± SEM); n = 7–8. (C) Spinal cords were harvested on day 14, and absolute cell numbers of spinal cord–infiltrating IL-17A–producing CD4+ T cells from the mice that received Pkm2-specific– or control shRNA–treated cells were evaluated by flow cytometry on day 14. Cumulative data are shown (mean ± SEM); n = 7–8. *P < 0.05; **P < 0.01 by 2-way ANOVA (A and B) or 2-tailed t test (C).