title: The NLRP3 inflammasome and bruton's tyrosine kinase in platelets co-regulate platelet activation, aggregation, and in vitro thrombus formation.
abstract: Cleavage of interleukin-1β (IL-1β) is a key inflammatory event in immune cells and platelets, which is mediated by nucleotide-binding domain leucine rich repeat containing protein (NLRP3)-dependent activation of caspase-1. In immune cells, NLRP3 and caspase-1 form inflammasome complexes with the adaptor proteins apoptosis-associated speck-like protein containing a CARD (ASC) and bruton's tyrosine kinase (BTK). In platelets, however, the regulatory triggers and the functional effects of the NLRP3 inflammasome are unknown. Here, we show in vitro that the platelet NLRP3 inflammasome contributes to platelet activation, aggregation, and thrombus formation. NLRP3 activity, as monitored by caspase-1 activation and cleavage and secretion of IL-1β, was upregulated in activated platelets, which was dependent on platelet BTK. Pharmacological inhibition or genetic ablation of BTK in platelets led to decreased platelet activation, aggregation, and in vitro thrombus formation. We identify a functionally relevant link between BTK and NLRP3 in platelets, with potential implications in disease states associated with abnormal coagulation and inflammation.
authors.affiliation: Department of Surgery, University of Pittsburgh, Pittsburgh, PA, USA.
authors.forename: Pranav
authors.initials: P
authors.lastname: Murthy
authors.affiliation: Department of Cardiology and Cardiovascular Diseases, Eberhard Karls University Tübingen, Germany.
authors.forename: Filip
authors.initials: F
authors.lastname: Durco
authors.affiliation: Department of Surgery, University of Pittsburgh, Pittsburgh, PA, USA.
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Miller-Ocuin
authors.affiliation: Department of Surgery, University of Pittsburgh, Pittsburgh, PA, USA.
authors.forename: Teiko
authors.initials: T
authors.lastname: Takedai
authors.affiliation: Department of Surgery, University of Pittsburgh, Pittsburgh, PA, USA.
authors.forename: Shruthi
authors.initials: S
authors.lastname: Shankar
authors.affiliation: Department of Surgery, University of Pittsburgh, Pittsburgh, PA, USA.
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: Liang
authors.affiliation: Interfaculty Institute for Cell Biology, Department of Immunology, Eberhard Karls University Tübingen, Germany.
authors.forename: Xiao
authors.initials: X
authors.lastname: Liu
authors.affiliation: Department of Surgery, University of Pittsburgh, Pittsburgh, PA, USA.
authors.forename: Xiangdong
authors.initials: X
authors.lastname: Cui
authors.affiliation: Department of Surgery, University of Pittsburgh, Pittsburgh, PA, USA.
authors.forename: Ulka
authors.initials: U
authors.lastname: Sachdev
authors.affiliation: Department of Cardiology and Cardiovascular Diseases, Eberhard Karls University Tübingen, Germany.
authors.forename: Dominik
authors.initials: D
authors.lastname: Rath
authors.affiliation: Department of Surgery, University of Pittsburgh, Pittsburgh, PA, USA.
authors.forename: Michael T
authors.initials: MT
authors.lastname: Lotze
authors.affiliation: Department of Surgery, University of Pittsburgh, Pittsburgh, PA, USA.
authors.forename: Herbert J
authors.initials: HJ
authors.lastname: Zeh
authors.affiliation: Department of Cardiology and Cardiovascular Diseases, Eberhard Karls University Tübingen, Germany.
authors.forename: Meinrad
authors.initials: M
authors.lastname: Gawaz
authors.affiliation: Interfaculty Institute for Cell Biology, Department of Immunology, Eberhard Karls University Tübingen, Germany.
authors.forename: Alexander N
authors.initials: AN
authors.lastname: Weber
authors.affiliation: Department of Cardiology and Cardiovascular Diseases, Eberhard Karls University Tübingen, Germany. Electronic address: sebastian.vogel@med.uni-tuebingen.de.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Vogel
mesh_terms: D000077329:Agammaglobulinaemia Tyrosine Kinase; D000818:Animals; D001549:Benzamides / Q000494:pharmacology; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000494:pharmacology; D002478:Cells, Cultured; D006801:Humans; D058847:Inflammasomes / Q000378:metabolism*; D053583:Interleukin-1beta / Q000378:metabolism; D008808:Mice, Inbred CBA; D018345:Mice, Knockout; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein / Q000235:genetics / Q000378:metabolism*; D009550:Nigericin / Q000494:pharmacology; D015539:Platelet Activation / Q000187:drug effects / Q000502:physiology*; D010974:Platelet Aggregation / Q000187:drug effects / Q000502:physiology; D011505:Protein-Tyrosine Kinases / Q000235:genetics / Q000378:metabolism*; D013927:Thrombosis / Q000378:metabolism*
chemical_list: D001549:Benzamides; D019086:Bridged Bicyclo Compounds, Heterocyclic; C555057:CGI 1746; D058847:Inflammasomes; D053583:Interleukin-1beta; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein; C492716:Nlrp3 protein, mouse; D011505:Protein-Tyrosine Kinases; D000077329:Agammaglobulinaemia Tyrosine Kinase; C000625949:BTK protein, human; C000625951:Btk protein, mouse; D009550:Nigericin
keywords: Aggregation; Bruton's tyrosine kinase; NLRP3; Platelets; Thrombosis