title: Human NACHT, LRR, and PYD domain-containing protein 3 (NLRP3) inflammasome activity is regulated by and potentially targetable through Bruton tyrosine kinase.
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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Interfaculty Institute for Cell Biology, Department of Immunology, University of Tübingen, Tübingen, Germany.
authors.forename: Xiao
authors.initials: X
authors.lastname: Liu
authors.affiliation: Interfaculty Institute for Cell Biology, Department of Immunology, University of Tübingen, Tübingen, Germany.
authors.forename: Tica
authors.initials: T
authors.lastname: Pichulik
authors.affiliation: Interfaculty Institute for Cell Biology, Department of Immunology, University of Tübingen, Tübingen, Germany.
authors.forename: Olaf-Oliver
authors.initials: OO
authors.lastname: Wolz
authors.affiliation: Interfaculty Institute for Cell Biology, Department of Immunology, University of Tübingen, Tübingen, Germany.
authors.forename: Truong-Minh
authors.initials: TM
authors.lastname: Dang
authors.affiliation: Institute of Innate Immunity, University Hospital Bonn, Bonn, Germany.
authors.forename: Andrea
authors.initials: A
authors.lastname: Stutz
authors.affiliation: Department of Dermatology, Johns Hopkins University, Baltimore, Md.
authors.forename: Carly
authors.initials: C
authors.lastname: Dillen
authors.affiliation: Interfaculty Institute for Cell Biology, Department of Immunology, University of Tübingen, Tübingen, Germany.
authors.forename: Magno
authors.initials: M
authors.lastname: Delmiro Garcia
authors.affiliation: Centre of Chronic Immunodeficiency, University Hospital Freiburg, Freiburg, Germany.
authors.forename: Helene
authors.initials: H
authors.lastname: Kraus
authors.affiliation: Interfaculty Institute for Cell Biology, Department of Immunology, University of Tübingen, Tübingen, Germany.
authors.forename: Sabine
authors.initials: S
authors.lastname: Dickhöfer
authors.affiliation: Interfaculty Institute of Microbiology and Infection Medicine, University of Tübingen, Tübingen, Germany.
authors.forename: Ellen
authors.initials: E
authors.lastname: Daiber
authors.affiliation: Interfaculty Institute of Microbiology and Infection Medicine, University of Tübingen, Tübingen, Germany.
authors.forename: Lisa
authors.initials: L
authors.lastname: Münzenmayer
authors.affiliation: Proteome Center Tübingen, University of Tübingen, Tübingen, Germany.
authors.forename: Silke
authors.initials: S
authors.lastname: Wahl
authors.affiliation: Department of Pediatrics I, University Hospital Tübingen, Tübingen, Germany.
authors.forename: Nikolaus
authors.initials: N
authors.lastname: Rieber
authors.affiliation: Department of Pediatrics I, University Hospital Tübingen, Tübingen, Germany.
authors.forename: Jasmin
authors.initials: J
authors.lastname: Kümmerle-Deschner
authors.affiliation: Department of Dermatology, University Hospital Tübingen, Tübingen, Germany.
authors.forename: Amir
authors.initials: A
authors.lastname: Yazdi
authors.affiliation: Proteome Center Tübingen, University of Tübingen, Tübingen, Germany.
authors.forename: Mirita
authors.initials: M
authors.lastname: Franz-Wachtel
authors.affiliation: Proteome Center Tübingen, University of Tübingen, Tübingen, Germany.
authors.forename: Boris
authors.initials: B
authors.lastname: Macek
authors.affiliation: Medical Hospital III, University Hospital Mainz, Mainz, Germany.
authors.forename: Markus
authors.initials: M
authors.lastname: Radsak
authors.affiliation: Department of Cardiology and Cardiovascular Diseases, University Hospital Tübingen, Tübingen, Germany.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Vogel
authors.affiliation: Interfaculty Institute of Microbiology and Infection Medicine, University of Tübingen, Tübingen, Germany.
authors.forename: Berit
authors.initials: B
authors.lastname: Schulte
authors.affiliation: Medical Hospital II (Department of Hematology and Oncology), University Hospital Tübingen, Tübingen, Germany.
authors.forename: Juliane Sarah
authors.initials: JS
authors.lastname: Walz
authors.affiliation: Department of Pediatrics I, University Hospital Tübingen, Tübingen, Germany.
authors.forename: Dominik
authors.initials: D
authors.lastname: Hartl
authors.affiliation: Institute of Innate Immunity, University Hospital Bonn, Bonn, Germany; Division of Infectious Diseases & Immunology, University of Massachusetts, Worcester, Mass.
authors.forename: Eicke
authors.initials: E
authors.lastname: Latz
authors.affiliation: Department of Internal Medicine III, Ulm University Hospital, Ulm, Germany.
authors.forename: Stephan
authors.initials: S
authors.lastname: Stilgenbauer
authors.affiliation: Centre of Chronic Immunodeficiency, University Hospital Freiburg, Freiburg, Germany.
authors.forename: Bodo
authors.initials: B
authors.lastname: Grimbacher
authors.affiliation: Department of Dermatology, Johns Hopkins University, Baltimore, Md.
authors.forename: Lloyd
authors.initials: L
authors.lastname: Miller
authors.affiliation: Department of Otorhinolaryngology, Ulm University Medical Center, Ulm, Germany.
authors.forename: Cornelia
authors.initials: C
authors.lastname: Brunner
authors.affiliation: Interfaculty Institute of Microbiology and Infection Medicine, University of Tübingen, Tübingen, Germany.
authors.forename: Christiane
authors.initials: C
authors.lastname: Wolz
authors.affiliation: Interfaculty Institute for Cell Biology, Department of Immunology, University of Tübingen, Tübingen, Germany. Electronic address: alexander.weber@uni.tuebingen.de.
authors.forename: Alexander N R
authors.initials: ANR
authors.lastname: Weber
mesh_terms: D056704:Adaptive Immunity; D048868:Adaptor Proteins, Signal Transducing; D000077329:Agammaglobulinaemia Tyrosine Kinase; D000361:Agammaglobulinemia / Q000235:genetics*; D000818:Animals; D051017:Apoptosis Regulatory Proteins; D001426:Bacterial Proteins / Q000276:immunology; D002478:Cells, Cultured; D056587:Cryopyrin-Associated Periodic Syndromes / Q000235:genetics*; D040181:Genetic Diseases, X-Linked / Q000235:genetics*; D006801:Humans; D007113:Immunity, Innate; D058847:Inflammasomes / Q000378:metabolism*; D007956:Leukocidins / Q000276:immunology; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D058990:Molecular Targeted Therapy; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein / Q000378:metabolism*; D000070576:NLR Proteins; D009550:Nigericin / Q000276:immunology; D011505:Protein-Tyrosine Kinases / Q000235:genetics / Q000378:metabolism*; D040901:Proteomics; D000071196:Pyrin Domain / Q000235:genetics; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism; D013203:Staphylococcal Infections / Q000276:immunology*; D013211:Staphylococcus aureus / Q000276:immunology*; D000097682:Lamin B Receptor
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D000077329:Agammaglobulinaemia Tyrosine Kinase; D051017:Apoptosis Regulatory Proteins; D001426:Bacterial Proteins; D058847:Inflammasomes; D007956:Leukocidins; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein; D000070576:NLR Proteins; D009550:Nigericin; D011505:Protein-Tyrosine Kinases; D018160:Receptors, Cytoplasmic and Nuclear; D000097682:Lamin B Receptor; C427903:NLRP1 protein, human; C453881:NLRP3 protein, human; C586430:leukocidin AB, Staphylococcus aureus; C000625949:BTK protein, human; C000625951:Btk protein, mouse
keywords: Bruton tyrosine kinase; IL-1; Muckle-Wells syndrome; NLRP3; Staphylococcus aureus; X-linked agammaglobulinemia; ibrutinib; inflammasome; inflammation; macrophage