Go to The Journal of Clinical Investigation
  • About
  • Editors
  • Consulting Editors
  • For authors
  • Publication ethics
  • Transfers
  • Advertising
  • Job board
  • Contact
  • Current issue
  • Past issues
  • By specialty
    • COVID-19
    • Cardiology
    • Immunology
    • Metabolism
    • Nephrology
    • Oncology
    • Pulmonology
    • All ...
  • Videos
  • Collections
    • Resource and Technical Advances
    • Clinical Medicine
    • Reviews
    • Editorials
    • Perspectives
    • Top read articles
  • JCI This Month
    • Current issue
    • Past issues

  • Current issue
  • Past issues
  • Specialties
  • In-Press Preview
  • Concise Communication
  • Editorials
  • Viewpoint
  • Top read articles
  • About
  • Editors
  • Consulting Editors
  • For authors
  • Publication ethics
  • Transfers
  • Advertising
  • Job board
  • Contact
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.
View: Text | PDF
Research Article Immunology

Pyruvate kinase M2 is requisite for Th1 and Th17 differentiation

  • Text
  • PDF
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

×

Figure 1

CaMK4 promotes glycolysis under ionomycin stimulation.

Options: View larger image (or click on image) Download as PowerPoint
CaMK4 promotes glycolysis under ionomycin stimulation.
(A and B) Camk4-s...
(A and B) Camk4-sufficient or -deficient naive CD4+ T cells were cultured with CD3 and CD28 Abs for 36 hours in vitro and extracellular acidification rate (ECAR) was analyzed by an extracellular flux analyzer. Glycolysis (A) and glycolytic capacity (B) in T cells from Camk4-sufficient or -deficient mice with or without ionomycin stimulation. Cumulative data of ECAR are shown (mean ± SEM); n = 4. (C and D) Camk4-sufficient naive CD4+ T cells were cultured with CD3 and CD28 Abs for 36 hours and after 1-hour pretreatment with or without CaMK4 inhibitor, KN-93, ECAR was measured. Glycolysis (C) and glycolytic capacity (D) in T cells with or without KN-93 (n = 4). Cumulative data of ECAR are shown (mean ± SEM); n = 4. *P < 0.05 by ANOVA. (E–H) Camk4-sufficient or -deficient naive CD4+ T cells were cultured under Th1- (E and F) or Th17-polarizing conditions (G and H) for 3 days in vitro and ECAR was analyzed by an extracellular flux analyzer. Glycolysis (E and G) and glycolytic capacity (F and H) in T cells from Camk4-sufficient or -deficient mice with ionomycin stimulation. Cumulative data of ECAR are shown (mean ± SEM); n = 4. *P < 0.05 by 2-tailed t test.

Copyright © 2023 American Society for Clinical Investigation
ISSN 2379-3708

Sign up for email alerts