cAbl Kinase Regulates Inflammasome Activation and Pyroptosis via ASC Phosphorylation

J Immunol. 2021 Mar 15;206(6):1329-1336. doi: 10.4049/jimmunol.2000969. Epub 2021 Feb 10.

Abstract

Inflammasome activation is regulated in part by the posttranslational modification of inflammasome proteins. Tyrosine phosphorylation is one possible modification. Having previously shown that the protein tyrosine kinase (PTK) inhibitor AG126 greatly inhibits inflammasome activation, we sought to uncover the target kinase. To do this, we screened a commercial tyrosine kinase library for inhibition of inflammasome-dependent IL-18/IL-1β release and pyroptosis. THP-1 cells (human monocyte cell line) were incubated with PTK inhibitors (0.1, 1, and 10 μM) before stimulation with LPS followed by ATP. The PTK inhibitors DCC-2036 (Rebastinib) and GZD824, specific for Bcr-Abl kinase, showed the most severe reduction of IL-18 and lactate dehydrogenase release at all concentrations used. The suggested kinase target, cAbl kinase, was then deleted in THP-1 cells by CRISPR/Cas9 editing and then tested for its role in inflammasome function and potential to phosphorylate the inflammasome adaptor ASC. The cABL knockout not only significantly inhibited inflammasome function but also decreased release of phosphorylated ASC after LPS/ATP stimulation. One predicted target of cAbl kinase is tyrosine 146 in ASC. Complementation of ASC knockout THP-1 cells with mutated Y146A ASC significantly abrogated inflammasome activation and ASC oligomerization as compared with wild-type ASC complementation. Thus, these findings support cAbl kinase as a positive regulator of inflammasome activity and pyroptosis, likely via phosphorylation of ASC.

Publication types

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

MeSH terms

  • Adenosine Triphosphate / immunology
  • Benzamides / pharmacology
  • CARD Signaling Adaptor Proteins / genetics
  • CARD Signaling Adaptor Proteins / metabolism*
  • Gene Knock-In Techniques
  • Gene Knockout Techniques
  • Humans
  • Inflammasomes / drug effects
  • Inflammasomes / immunology*
  • Inflammasomes / metabolism
  • Lipopolysaccharides / immunology
  • Mutation
  • Phosphorylation / drug effects
  • Phosphorylation / genetics
  • Phosphorylation / immunology
  • Protein Kinase Inhibitors / pharmacology*
  • Proto-Oncogene Proteins c-abl / genetics
  • Proto-Oncogene Proteins c-abl / metabolism*
  • Pyrazoles / pharmacology
  • Pyridines / pharmacology
  • Pyroptosis / drug effects
  • Pyroptosis / immunology*
  • Quinolines / pharmacology
  • THP-1 Cells
  • Tyrphostins / pharmacology

Substances

  • 3-((1H-pyrazolo(3,4-b)pyridin-5-yl)ethynyl)-4-methyl-N-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)benzamide
  • Benzamides
  • CARD Signaling Adaptor Proteins
  • Inflammasomes
  • Lipopolysaccharides
  • PYCARD protein, human
  • Protein Kinase Inhibitors
  • Pyrazoles
  • Pyridines
  • Quinolines
  • Tyrphostins
  • AG 127
  • rebastinib
  • Adenosine Triphosphate
  • ABL1 protein, human
  • Proto-Oncogene Proteins c-abl