Tryptophan potentiates CD8+ T cells against cancer cells by TRIP12 tryptophanylation and surface PD-1 downregulation

J Immunother Cancer. 2021 Jul;9(7):e002840. doi: 10.1136/jitc-2021-002840.

Abstract

Background: Tryptophan catabolites suppress immunity. Therefore, blocking tryptophan catabolism with indoleamine 2,3-dioxygenase (IDO) inhibitors is pursued as an anticancer strategy.

Methods: The intracellular level of tryptophan and kynurenine was detected by mass spectrum analysis. The effect of tryptophan and IDO inhibitors on cell surface programmed cell death protein 1 (PD-1) level were measured by flow cytometry. A set of biochemical analyses were used to figure out the underlying mechanism. In vitro co-culture system, syngeneic mouse models, immunofluorescent staining, and flow cytometry analysis were employed to investigate the role of tryptophan and IDO inhibitor in regulating the cytotoxicity of CD8+ T cells.

Results: Here, we reported that IDO inhibitors activated CD8+ T cells also by accumulating tryptophan that downregulated PD-1. Tryptophan and IDO inhibitors administration, both increased intracellular tryptophan, and tryptophanyl-tRNA synthetase (WARS) overexpression decreased Jurkat and mice CD8+ T cell surface PD-1. Mechanistically, WARS tryptophanylated lysine 1136 of and activated E3 ligase TRIP12 to degrade NFATc1, a PD-1 transcription activator. SIRT1 de-tryptophanylated TRIP12 and reversed the effects of tryptophan and WARS on PD-1. Tryptophan or IDO inhibitors potentiated CD8+ T cells to induce apoptosis of co-cultured cancer cells, increased cancer-infiltrating CD8+ T cells and slowed down tumor growth of lung cancer in mice.

Conclusions: Our results revealed the immune-activating efficacy of tryptophan, and suggested tryptophan supplemental may benefit IDO inhibitors and PD-1 blockade during anticancer treatments.

Keywords: CD8-positive T-lymphocytes; immunotherapy; indoleamine 2,3-dioxygenase.

MeSH terms

  • Animals
  • CD8-Positive T-Lymphocytes / metabolism*
  • Carrier Proteins / metabolism*
  • Cell Proliferation
  • Down-Regulation
  • Female
  • Humans
  • Mice
  • Programmed Cell Death 1 Receptor / metabolism*
  • Transfection
  • Tryptophan / pharmacology
  • Tryptophan / therapeutic use*
  • Ubiquitin-Protein Ligases / metabolism*

Substances

  • Carrier Proteins
  • Programmed Cell Death 1 Receptor
  • Tryptophan
  • TRIP12 protein, human
  • Ubiquitin-Protein Ligases