Neutrophil elastase activates protease-activated receptor-2 (PAR2) and transient receptor potential vanilloid 4 (TRPV4) to cause inflammation and pain

Peishen Zhao, Tina Marie Lieu, Nicholas Barlow, Silvia Sostegni, Silke Haerteis, Christoph Korbmacher, Wolfgang Liedtke, Nestor N Jimenez-Vargas, Stephen Vanner, Nigel William Bunnett

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Proteases that cleave protease-activated receptor-2 (PAR2)at Arg36 ? Ser37 reveal a tethered ligand that binds to the cleaved receptor. PAR2 activates transient receptor potential (TRP) channels of nociceptive neurons to induce neurogenic inflammation and pain. Although proteases that cleave PAR2 at noncanonical sites can trigger distinct signaling cascades, the functional importance of the PAR2-biased agonism is uncertain. We investigated whether neutrophil elastase, a biased agonist of PAR2, causes inflammation and pain by activating PAR2 and TRP vanilloid 4 (TRPV4). Elastase cleaved human PAR2 at Ala66 ? Ser67 and Ser67 ? Val68. Elastasestimulated PAR2-dependent cAMP accumulation and ERK1/2 activation, but not Ca2+ mobilization, in KNRK cells. Elastase induced PAR2 coupling to Goas but not Goaq in HEK293 cells. Although elastase did not promote recruitment of G protein-coupled receptor kinase-2 (GRK2) or ?-arrestin to PAR2, consistent with its inability to promote receptor endocytosis, elastase did stimulate GRK6 recruitment. Elastase caused PAR2-dependent sensitization of TRPV4 currents in Xenopus laevis oocytes by adenylyl cyclaseand protein kinase A (PKA)-dependent mechanisms. Elastase stimulated PAR2-dependent cAMP formationand ERK1/2 phosphorylation, and a PAR2- and TRPV4-mediated influx of extracellular Ca2+ in mouse nociceptors. Adenylyl cyclase and PKA-mediated elastase-induced activation of TRPV4 and hyperexcitability of nociceptors. Intraplantar injection of elastase to mice caused edema and mechanical hyperalgesia by PAR2- and TRPV4-mediated mechanisms. Thus, the elastasebiased agonism of PAR2 causes Gas-dependent activation of adenylyl cyclase and PKA, which activates TRPV4 and sensitizes nociceptors to cause inflammation and pain. Our results identify a novel mechanism of elastase-induced activation of TRPV4 and expand the role of PAR2 as a mediator of protease-driven inflammation and pain.
Original languageEnglish
Pages (from-to)13875 - 13887
Number of pages13
JournalJournal of Biological Chemistry
Volume290
Issue number22
DOIs
Publication statusPublished - 2015

Cite this

Zhao, Peishen ; Lieu, Tina Marie ; Barlow, Nicholas ; Sostegni, Silvia ; Haerteis, Silke ; Korbmacher, Christoph ; Liedtke, Wolfgang ; Jimenez-Vargas, Nestor N ; Vanner, Stephen ; Bunnett, Nigel William. / Neutrophil elastase activates protease-activated receptor-2 (PAR2) and transient receptor potential vanilloid 4 (TRPV4) to cause inflammation and pain. In: Journal of Biological Chemistry. 2015 ; Vol. 290, No. 22. pp. 13875 - 13887.
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title = "Neutrophil elastase activates protease-activated receptor-2 (PAR2) and transient receptor potential vanilloid 4 (TRPV4) to cause inflammation and pain",
abstract = "Proteases that cleave protease-activated receptor-2 (PAR2)at Arg36 ? Ser37 reveal a tethered ligand that binds to the cleaved receptor. PAR2 activates transient receptor potential (TRP) channels of nociceptive neurons to induce neurogenic inflammation and pain. Although proteases that cleave PAR2 at noncanonical sites can trigger distinct signaling cascades, the functional importance of the PAR2-biased agonism is uncertain. We investigated whether neutrophil elastase, a biased agonist of PAR2, causes inflammation and pain by activating PAR2 and TRP vanilloid 4 (TRPV4). Elastase cleaved human PAR2 at Ala66 ? Ser67 and Ser67 ? Val68. Elastasestimulated PAR2-dependent cAMP accumulation and ERK1/2 activation, but not Ca2+ mobilization, in KNRK cells. Elastase induced PAR2 coupling to Goas but not Goaq in HEK293 cells. Although elastase did not promote recruitment of G protein-coupled receptor kinase-2 (GRK2) or ?-arrestin to PAR2, consistent with its inability to promote receptor endocytosis, elastase did stimulate GRK6 recruitment. Elastase caused PAR2-dependent sensitization of TRPV4 currents in Xenopus laevis oocytes by adenylyl cyclaseand protein kinase A (PKA)-dependent mechanisms. Elastase stimulated PAR2-dependent cAMP formationand ERK1/2 phosphorylation, and a PAR2- and TRPV4-mediated influx of extracellular Ca2+ in mouse nociceptors. Adenylyl cyclase and PKA-mediated elastase-induced activation of TRPV4 and hyperexcitability of nociceptors. Intraplantar injection of elastase to mice caused edema and mechanical hyperalgesia by PAR2- and TRPV4-mediated mechanisms. Thus, the elastasebiased agonism of PAR2 causes Gas-dependent activation of adenylyl cyclase and PKA, which activates TRPV4 and sensitizes nociceptors to cause inflammation and pain. Our results identify a novel mechanism of elastase-induced activation of TRPV4 and expand the role of PAR2 as a mediator of protease-driven inflammation and pain.",
author = "Peishen Zhao and Lieu, {Tina Marie} and Nicholas Barlow and Silvia Sostegni and Silke Haerteis and Christoph Korbmacher and Wolfgang Liedtke and Jimenez-Vargas, {Nestor N} and Stephen Vanner and Bunnett, {Nigel William}",
year = "2015",
doi = "10.1074/jbc.M115.642736",
language = "English",
volume = "290",
pages = "13875 -- 13887",
journal = "Journal of Biological Chemistry",
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Neutrophil elastase activates protease-activated receptor-2 (PAR2) and transient receptor potential vanilloid 4 (TRPV4) to cause inflammation and pain. / Zhao, Peishen; Lieu, Tina Marie; Barlow, Nicholas; Sostegni, Silvia; Haerteis, Silke; Korbmacher, Christoph; Liedtke, Wolfgang; Jimenez-Vargas, Nestor N; Vanner, Stephen; Bunnett, Nigel William.

In: Journal of Biological Chemistry, Vol. 290, No. 22, 2015, p. 13875 - 13887.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Neutrophil elastase activates protease-activated receptor-2 (PAR2) and transient receptor potential vanilloid 4 (TRPV4) to cause inflammation and pain

AU - Zhao, Peishen

AU - Lieu, Tina Marie

AU - Barlow, Nicholas

AU - Sostegni, Silvia

AU - Haerteis, Silke

AU - Korbmacher, Christoph

AU - Liedtke, Wolfgang

AU - Jimenez-Vargas, Nestor N

AU - Vanner, Stephen

AU - Bunnett, Nigel William

PY - 2015

Y1 - 2015

N2 - Proteases that cleave protease-activated receptor-2 (PAR2)at Arg36 ? Ser37 reveal a tethered ligand that binds to the cleaved receptor. PAR2 activates transient receptor potential (TRP) channels of nociceptive neurons to induce neurogenic inflammation and pain. Although proteases that cleave PAR2 at noncanonical sites can trigger distinct signaling cascades, the functional importance of the PAR2-biased agonism is uncertain. We investigated whether neutrophil elastase, a biased agonist of PAR2, causes inflammation and pain by activating PAR2 and TRP vanilloid 4 (TRPV4). Elastase cleaved human PAR2 at Ala66 ? Ser67 and Ser67 ? Val68. Elastasestimulated PAR2-dependent cAMP accumulation and ERK1/2 activation, but not Ca2+ mobilization, in KNRK cells. Elastase induced PAR2 coupling to Goas but not Goaq in HEK293 cells. Although elastase did not promote recruitment of G protein-coupled receptor kinase-2 (GRK2) or ?-arrestin to PAR2, consistent with its inability to promote receptor endocytosis, elastase did stimulate GRK6 recruitment. Elastase caused PAR2-dependent sensitization of TRPV4 currents in Xenopus laevis oocytes by adenylyl cyclaseand protein kinase A (PKA)-dependent mechanisms. Elastase stimulated PAR2-dependent cAMP formationand ERK1/2 phosphorylation, and a PAR2- and TRPV4-mediated influx of extracellular Ca2+ in mouse nociceptors. Adenylyl cyclase and PKA-mediated elastase-induced activation of TRPV4 and hyperexcitability of nociceptors. Intraplantar injection of elastase to mice caused edema and mechanical hyperalgesia by PAR2- and TRPV4-mediated mechanisms. Thus, the elastasebiased agonism of PAR2 causes Gas-dependent activation of adenylyl cyclase and PKA, which activates TRPV4 and sensitizes nociceptors to cause inflammation and pain. Our results identify a novel mechanism of elastase-induced activation of TRPV4 and expand the role of PAR2 as a mediator of protease-driven inflammation and pain.

AB - Proteases that cleave protease-activated receptor-2 (PAR2)at Arg36 ? Ser37 reveal a tethered ligand that binds to the cleaved receptor. PAR2 activates transient receptor potential (TRP) channels of nociceptive neurons to induce neurogenic inflammation and pain. Although proteases that cleave PAR2 at noncanonical sites can trigger distinct signaling cascades, the functional importance of the PAR2-biased agonism is uncertain. We investigated whether neutrophil elastase, a biased agonist of PAR2, causes inflammation and pain by activating PAR2 and TRP vanilloid 4 (TRPV4). Elastase cleaved human PAR2 at Ala66 ? Ser67 and Ser67 ? Val68. Elastasestimulated PAR2-dependent cAMP accumulation and ERK1/2 activation, but not Ca2+ mobilization, in KNRK cells. Elastase induced PAR2 coupling to Goas but not Goaq in HEK293 cells. Although elastase did not promote recruitment of G protein-coupled receptor kinase-2 (GRK2) or ?-arrestin to PAR2, consistent with its inability to promote receptor endocytosis, elastase did stimulate GRK6 recruitment. Elastase caused PAR2-dependent sensitization of TRPV4 currents in Xenopus laevis oocytes by adenylyl cyclaseand protein kinase A (PKA)-dependent mechanisms. Elastase stimulated PAR2-dependent cAMP formationand ERK1/2 phosphorylation, and a PAR2- and TRPV4-mediated influx of extracellular Ca2+ in mouse nociceptors. Adenylyl cyclase and PKA-mediated elastase-induced activation of TRPV4 and hyperexcitability of nociceptors. Intraplantar injection of elastase to mice caused edema and mechanical hyperalgesia by PAR2- and TRPV4-mediated mechanisms. Thus, the elastasebiased agonism of PAR2 causes Gas-dependent activation of adenylyl cyclase and PKA, which activates TRPV4 and sensitizes nociceptors to cause inflammation and pain. Our results identify a novel mechanism of elastase-induced activation of TRPV4 and expand the role of PAR2 as a mediator of protease-driven inflammation and pain.

UR - http://www.jbc.org/content/290/22/13875.full.pdf+html

U2 - 10.1074/jbc.M115.642736

DO - 10.1074/jbc.M115.642736

M3 - Article

VL - 290

SP - 13875

EP - 13887

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 1083-351X

IS - 22

ER -