Pyruvate kinase M2 is requisite for Th1 and Th17 differentiation

JCI Insight. 2019 Jun 20;4(12):e127395. doi: 10.1172/jci.insight.127395.

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.

Keywords: Autoimmune diseases; Autoimmunity; Immunology; T cells.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Calcium-Calmodulin-Dependent Protein Kinase Type 4 / metabolism*
  • Carrier Proteins / antagonists & inhibitors
  • Carrier Proteins / metabolism*
  • Dimethyl Sulfoxide / pharmacology
  • Encephalomyelitis, Autoimmune, Experimental / enzymology
  • Encephalomyelitis, Autoimmune, Experimental / immunology
  • Enzyme Inhibitors / pharmacology
  • Glycolysis
  • Lymphopoiesis*
  • Membrane Proteins / antagonists & inhibitors
  • Membrane Proteins / metabolism*
  • Mice, Inbred C57BL
  • Naphthoquinones / pharmacology
  • Th1 Cells / drug effects
  • Th1 Cells / enzymology*
  • Th1 Cells / physiology
  • Th17 Cells / drug effects
  • Th17 Cells / enzymology*
  • Th17 Cells / physiology
  • Thyroid Hormones / metabolism*

Substances

  • Carrier Proteins
  • Enzyme Inhibitors
  • Membrane Proteins
  • Naphthoquinones
  • Thyroid Hormones
  • thyroid hormone-binding proteins
  • shikonin
  • CAMK4 protein, human
  • Calcium-Calmodulin-Dependent Protein Kinase Type 4
  • Dimethyl Sulfoxide