Xiao Liu1, Tica Pichulik1, Olaf-Oliver Wolz1, Truong-Minh Dang1, Andrea Stutz2, Carly Dillen3, Magno Delmiro Garcia1, Helene Kraus4, Sabine Dickhöfer1, Ellen Daiber5, Lisa Münzenmayer5, Silke Wahl6, Nikolaus Rieber7, Jasmin Kümmerle-Deschner7, Amir Yazdi8, Mirita Franz-Wachtel6, Boris Macek6, Markus Radsak9, Sebastian Vogel10, Berit Schulte5, Juliane Sarah Walz11, Dominik Hartl7, Eicke Latz12, Stephan Stilgenbauer13, Bodo Grimbacher4, Lloyd Miller3, Cornelia Brunner14, Christiane Wolz5, Alexander N R Weber15. 1. Interfaculty Institute for Cell Biology, Department of Immunology, University of Tübingen, Tübingen, Germany. 2. Institute of Innate Immunity, University Hospital Bonn, Bonn, Germany. 3. Department of Dermatology, Johns Hopkins University, Baltimore, Md. 4. Centre of Chronic Immunodeficiency, University Hospital Freiburg, Freiburg, Germany. 5. Interfaculty Institute of Microbiology and Infection Medicine, University of Tübingen, Tübingen, Germany. 6. Proteome Center Tübingen, University of Tübingen, Tübingen, Germany. 7. Department of Pediatrics I, University Hospital Tübingen, Tübingen, Germany. 8. Department of Dermatology, University Hospital Tübingen, Tübingen, Germany. 9. Medical Hospital III, University Hospital Mainz, Mainz, Germany. 10. Department of Cardiology and Cardiovascular Diseases, University Hospital Tübingen, Tübingen, Germany. 11. Medical Hospital II (Department of Hematology and Oncology), University Hospital Tübingen, Tübingen, Germany. 12. Institute of Innate Immunity, University Hospital Bonn, Bonn, Germany; Division of Infectious Diseases & Immunology, University of Massachusetts, Worcester, Mass. 13. Department of Internal Medicine III, Ulm University Hospital, Ulm, Germany. 14. Department of Otorhinolaryngology, Ulm University Medical Center, Ulm, Germany. 15. Interfaculty Institute for Cell Biology, Department of Immunology, University of Tübingen, Tübingen, Germany. Electronic address: alexander.weber@uni.tuebingen.de.
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.
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 NLRP3inflammasome-linked inflammation could potentially be targeted pharmacologically through BTK.
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