Human NACHT, LRR, and PYD Domain-Containing Protein 3 (NLRP3) Inflammasome Activity Is Regulated by and Potentially Targetable Through Bruton Tyrosine Kinase

J Allergy Clin Immunol. 2017 Oct;140(4):1054-1067.e10. doi: 10.1016/j.jaci.2017.01.017. Epub 2017 Feb 16.

Abstract

Background: The Nod-like receptor NACHT, LRR, and PYD domain-containing protein 3 (NLRP3) and Bruton tyrosine kinase (BTK) are protagonists in innate and adaptive immunity, respectively. NLRP3 senses exogenous and endogenous insults, leading to inflammasome activation, which occurs spontaneously in patients with Muckle-Wells syndrome; BTK mutations cause the genetic immunodeficiency X-linked agammaglobulinemia (XLA). However, to date, few proteins that regulate NLRP3 inflammasome activity in human primary immune cells have been identified, and clinically promising pharmacologic targeting strategies remain elusive.

Objective: We sought to identify novel regulators of the NLRP3 inflammasome in human cells with a view to exploring interference with inflammasome activity at the level of such regulators.

Methods: After proteome-wide phosphoproteomics, the identified novel regulator BTK was studied in human and murine cells by using pharmacologic and genetic BTK ablation.

Results: Here we show that BTK is a critical regulator of NLRP3 inflammasome activation: pharmacologic (using the US Food and Drug Administration-approved inhibitor ibrutinib) and genetic (in patients with XLA and Btk knockout mice) BTK ablation in primary immune cells led to reduced IL-1β processing and secretion in response to nigericin and the Staphylococcus aureus toxin leukocidin AB (LukAB). BTK affected apoptosis-associated speck-like protein containing a CARD (ASC) speck formation and caspase-1 cleavage and interacted with NLRP3 and ASC. S aureus infection control in vivo and IL-1β release from cells of patients with Muckle-Wells syndrome were impaired by ibrutinib. Notably, IL-1β processing and release from immune cells isolated from patients with cancer receiving ibrutinib therapy were reduced.

Conclusion: Our data suggest that XLA might result in part from genetic inflammasome deficiency and that NLRP3 inflammasome-linked inflammation could potentially be targeted pharmacologically through BTK.

Keywords: Bruton tyrosine kinase; IL-1; Muckle-Wells syndrome; NLRP3; Staphylococcus aureus; X-linked agammaglobulinemia; ibrutinib; inflammasome; inflammation; macrophage.

MeSH terms

  • Adaptive Immunity
  • Adaptor Proteins, Signal Transducing
  • Agammaglobulinaemia Tyrosine Kinase
  • Agammaglobulinemia / genetics*
  • Animals
  • Apoptosis Regulatory Proteins
  • Bacterial Proteins / immunology
  • Cells, Cultured
  • Cryopyrin-Associated Periodic Syndromes / genetics*
  • Genetic Diseases, X-Linked / genetics*
  • Humans
  • Immunity, Innate
  • Inflammasomes / metabolism*
  • Leukocidins / immunology
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Molecular Targeted Therapy
  • NLR Family, Pyrin Domain-Containing 3 Protein / metabolism*
  • Nigericin / immunology
  • Protein-Tyrosine Kinases / genetics
  • Protein-Tyrosine Kinases / metabolism*
  • Proteomics
  • Pyrin Domain / genetics
  • Receptors, Cytoplasmic and Nuclear / metabolism
  • Staphylococcal Infections / immunology*
  • Staphylococcus aureus / immunology*

Substances

  • Adaptor Proteins, Signal Transducing
  • Apoptosis Regulatory Proteins
  • Bacterial Proteins
  • Inflammasomes
  • Leukocidins
  • NLR Family, Pyrin Domain-Containing 3 Protein
  • NLRP1 protein, human
  • NLRP3 protein, human
  • Receptors, Cytoplasmic and Nuclear
  • lamin B receptor
  • leukocidin AB, Staphylococcus aureus
  • Protein-Tyrosine Kinases
  • Agammaglobulinaemia Tyrosine Kinase
  • BTK protein, human
  • Btk protein, mouse
  • Nigericin

Supplementary concepts

  • Bruton type agammaglobulinemia