title: Bimodal role of NADPH oxidases in the regulation of biglycan-triggered IL-1β synthesis.
abstract: Biglycan, a ubiquitous proteoglycan, acts as a danger signal when released from the extracellular matrix. As such, biglycan triggers the synthesis and maturation of interleukin-1β (IL-1β) in a Toll-like receptor (TLR) 2-, TLR4-, and reactive oxygen species (ROS)-dependent manner. Here, we discovered that biglycan autonomously regulates the balance in IL-1β production in vitro and in vivo by modulating expression, activity and stability of NADPH oxidase (NOX) 1, 2 and 4 enzymes via different TLR pathways. In primary murine macrophages, biglycan triggered NOX1/4-mediated ROS generation, thereby enhancing IL-1β expression. Surprisingly, biglycan inhibited IL-1β due to enhancement of NOX2 synthesis and activation, by selectively interacting with TLR4. Synthesis of NOX2 was mediated by adaptor molecule Toll/IL-1R domain-containing adaptor inducing IFN-β (TRIF). Via myeloid differentiation primary response protein (MyD88) as well as Rac1 activation and Erk phosphorylation, biglycan triggered translocation of the cytosolic NOX2 subunit p47(phox) to the plasma membrane, an obligatory step for NOX2 activation. In contrast, by engaging TLR2, soluble biglycan stimulated the expression of heat shock protein (HSP) 70, which bound to NOX2, and consequently impaired the inhibitory function of NOX2 on IL-1β expression. Notably, a genetic background lacking biglycan reduced HSP70 expression, rescued the enhanced renal IL-1β production and improved kidney function of Nox2(-/y) mice in a model of renal ischemia reperfusion injury. Here, we provide a novel mechanism where the danger molecule biglycan influences NOX2 synthesis and activation via different TLR pathways, thereby regulating inflammation severity. Thus, selective inhibition of biglycan-TLR2 or biglycan-TLR4 signaling could be a novel therapeutic approach in ROS-mediated inflammatory diseases.
authors.affiliation: Pharmazentrum Frankfurt, Institut für Allgemeine Pharmakologie und Toxikologie, Klinikum der Goethe-Universität Frankfurt am Main, Frankfurt am Main, Germany.
authors.forename: Louise Tzung-Harn
authors.initials: LT
authors.lastname: Hsieh
authors.affiliation: Pharmazentrum Frankfurt, Institut für Allgemeine Pharmakologie und Toxikologie, Klinikum der Goethe-Universität Frankfurt am Main, Frankfurt am Main, Germany.
authors.forename: Helena
authors.initials: H
authors.lastname: Frey
authors.affiliation: Pharmazentrum Frankfurt, Institut für Allgemeine Pharmakologie und Toxikologie, Klinikum der Goethe-Universität Frankfurt am Main, Frankfurt am Main, Germany; Department of Pharmacology, National Institute for Chemical-Pharmaceutical Research and Development, 112 Vitan Avenue, Bucharest, Romania.
authors.forename: Madalina-Viviana
authors.initials: MV
authors.lastname: Nastase
authors.affiliation: Pharmazentrum Frankfurt, Institut für Allgemeine Pharmakologie und Toxikologie, Klinikum der Goethe-Universität Frankfurt am Main, Frankfurt am Main, Germany.
authors.forename: Claudia
authors.initials: C
authors.lastname: Tredup
authors.affiliation: Pharmazentrum Frankfurt, Institut für Allgemeine Pharmakologie und Toxikologie, Klinikum der Goethe-Universität Frankfurt am Main, Frankfurt am Main, Germany.
authors.forename: Adrian
authors.initials: A
authors.lastname: Hoffmann
authors.affiliation: Pharmazentrum Frankfurt, Institut für Allgemeine Pharmakologie und Toxikologie, Klinikum der Goethe-Universität Frankfurt am Main, Frankfurt am Main, Germany.
authors.forename: Chiara
authors.initials: C
authors.lastname: Poluzzi
authors.affiliation: Pharmazentrum Frankfurt, Institut für Allgemeine Pharmakologie und Toxikologie, Klinikum der Goethe-Universität Frankfurt am Main, Frankfurt am Main, Germany.
authors.forename: Jinyang
authors.initials: J
authors.lastname: Zeng-Brouwers
authors.affiliation: Pharmazentrum Frankfurt, Institut für Allgemeine Pharmakologie und Toxikologie, Klinikum der Goethe-Universität Frankfurt am Main, Frankfurt am Main, Germany.
authors.forename: Tina
authors.initials: T
authors.lastname: Manon-Jensen
authors.affiliation: Institut für Kardiovaskulare Physiologie, Klinikum der Goethe-Universität Frankfurt am Main, Frankfurt am Main, Germany.
authors.forename: Katrin
authors.initials: K
authors.lastname: Schröder
authors.affiliation: Institut für Kardiovaskulare Physiologie, Klinikum der Goethe-Universität Frankfurt am Main, Frankfurt am Main, Germany.
authors.forename: Ralf P
authors.initials: RP
authors.lastname: Brandes
authors.affiliation: Department of Pathology, Anatomy and Cell Biology and the Cancer Cell Biology and Signaling Program, Kimmel Cancer Center, Sidney Kimmel Medical College at Thomas Jefferson University, Philadelphia, PA 19107, USA.
authors.forename: Renato V
authors.initials: RV
authors.lastname: Iozzo
authors.affiliation: Pharmazentrum Frankfurt, Institut für Allgemeine Pharmakologie und Toxikologie, Klinikum der Goethe-Universität Frankfurt am Main, Frankfurt am Main, Germany. Electronic address: schaefer@med.uni-frankfurt.de.
authors.forename: Liliana
authors.initials: L
authors.lastname: Schaefer
mesh_terms: D000818:Animals; D058578:Biglycan / Q000378:metabolism*; D002478:Cells, Cultured; D016209:Interleukin-8 / Q000378:metabolism*; D007668:Kidney / Q000293:injuries / Q000378:metabolism; D008264:Macrophages / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000378:metabolism*; D051379:Mice; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism*; D000074624:NADPH Oxidase 1; D000074662:NADPH Oxidase 2; D000074663:NADPH Oxidase 4; D019255:NADPH Oxidases / Q000378:metabolism*; D015427:Reperfusion Injury / Q000209:etiology; D015398:Signal Transduction; D051193:Toll-Like Receptors / Q000378:metabolism
chemical_list: C546884:BGN protein, human; D058578:Biglycan; D016209:Interleukin-8; D008562:Membrane Glycoproteins; D051193:Toll-Like Receptors; D009247:NADH, NADPH Oxidoreductases; C488842:Cybb protein, mouse; D000074624:NADPH Oxidase 1; D000074662:NADPH Oxidase 2; D000074663:NADPH Oxidase 4; D019255:NADPH Oxidases; C000614711:NOX1 protein, mouse; C422943:Nox4 protein, mouse
keywords: Extracellular matrix; Inflammation; Reactive oxygen species (ROS); Small leucine-rich proteoglycan; Toll-like receptor