Protease-activated receptor-2 in endosomes signals persistent pain of irritable bowel syndrome

Nestor N. Jimenez-Vargas, Luke A. Pattison, Peishen Zhao, TinaMarie Lieu, Rocco Latorre, Dane D. Jensen, Joel Castro, Luigi Aurelio, Giang T. Le, Bernard Flynn, Carmen Klein Herenbrink, Holly R. Yeatman, Laura Edgington-Mitchell, Christopher J.H. Porter, Michelle L. Halls, Meritxell Canals, Nicholas A. Veldhuis, Daniel P. Poole, Peter McLean, Gareth A. Hicks & 7 others Nicole Scheff, Elyssa Chen, Aditi Bhattacharya, Brian L. Schmidt, Stuart M. Brierley, Stephen J. Vanner, Nigel W. Bunnett

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Once activated at the surface of cells, G protein-coupled receptors (GPCRs) redistribute to endosomes, where they can continue to signal. Whether GPCRs in endosomes generate signals that contribute to human disease is unknown. We evaluated endosomal signaling of protease-activated receptor-2 (PAR2), which has been proposed to mediate pain in patients with irritable bowel syndrome (IBS). Trypsin, elastase, and cathepsin S, which are activated in the colonic mucosa of patients with IBS and in experimental animals with colitis, caused persistent PAR2-dependent hyperexcitability of nociceptors, sensitization of colonic afferent neurons to mechanical stimuli, and somatic mechanical allodynia. Inhibitors of clathrin- and dynamin-dependent endocytosis and of mitogen-activated protein kinase kinase-1 prevented trypsin-induced hyperexcitability, sensitization, and allodynia. However, they did not affect elastase- or cathepsin S-induced hyperexcitability, sensitization, or allodynia. Trypsin stimulated endocytosis of PAR2, which signaled from endosomes to activate extracellular signal-regulated kinase. Elastase and cathepsin S did not stimulate endocytosis of PAR2, which signaled from the plasma membrane to activate adenylyl cyclase. Biopsies of colonic mucosa from IBS patients released proteases that induced persistent PAR2-dependent hyperexcitability of nociceptors, and PAR2 association with β-arrestins, which mediate endocytosis. Conjugation to cholestanol promoted delivery and retention of antagonists in endosomes containing PAR2. A cholestanol-conjugated PAR2 antagonist prevented persistent trypsin- and IBS protease-induced hyperexcitability of nociceptors. The results reveal that PAR2 signaling from endosomes underlies the persistent hyperexcitability of nociceptors that mediates chronic pain of IBS. Endosomally targeted PAR2 antagonists are potential therapies for IBS pain. GPCRs in endosomes transmit signals that contribute to human diseases.

Original languageEnglish
Pages (from-to)E7438-E7447
Number of pages10
JournalProceedings of the National Academy of Sciences of the United States of America
Volume115
Issue number31
DOIs
Publication statusPublished - 31 Jul 2018

Keywords

  • Endosomes
  • Pain
  • Proteases
  • Receptors

Cite this

Jimenez-Vargas, Nestor N. ; Pattison, Luke A. ; Zhao, Peishen ; Lieu, TinaMarie ; Latorre, Rocco ; Jensen, Dane D. ; Castro, Joel ; Aurelio, Luigi ; Le, Giang T. ; Flynn, Bernard ; Herenbrink, Carmen Klein ; Yeatman, Holly R. ; Edgington-Mitchell, Laura ; Porter, Christopher J.H. ; Halls, Michelle L. ; Canals, Meritxell ; Veldhuis, Nicholas A. ; Poole, Daniel P. ; McLean, Peter ; Hicks, Gareth A. ; Scheff, Nicole ; Chen, Elyssa ; Bhattacharya, Aditi ; Schmidt, Brian L. ; Brierley, Stuart M. ; Vanner, Stephen J. ; Bunnett, Nigel W. / Protease-activated receptor-2 in endosomes signals persistent pain of irritable bowel syndrome. In: Proceedings of the National Academy of Sciences of the United States of America. 2018 ; Vol. 115, No. 31. pp. E7438-E7447.
@article{acada9a59e2f47119cc537b80ca11509,
title = "Protease-activated receptor-2 in endosomes signals persistent pain of irritable bowel syndrome",
abstract = "Once activated at the surface of cells, G protein-coupled receptors (GPCRs) redistribute to endosomes, where they can continue to signal. Whether GPCRs in endosomes generate signals that contribute to human disease is unknown. We evaluated endosomal signaling of protease-activated receptor-2 (PAR2), which has been proposed to mediate pain in patients with irritable bowel syndrome (IBS). Trypsin, elastase, and cathepsin S, which are activated in the colonic mucosa of patients with IBS and in experimental animals with colitis, caused persistent PAR2-dependent hyperexcitability of nociceptors, sensitization of colonic afferent neurons to mechanical stimuli, and somatic mechanical allodynia. Inhibitors of clathrin- and dynamin-dependent endocytosis and of mitogen-activated protein kinase kinase-1 prevented trypsin-induced hyperexcitability, sensitization, and allodynia. However, they did not affect elastase- or cathepsin S-induced hyperexcitability, sensitization, or allodynia. Trypsin stimulated endocytosis of PAR2, which signaled from endosomes to activate extracellular signal-regulated kinase. Elastase and cathepsin S did not stimulate endocytosis of PAR2, which signaled from the plasma membrane to activate adenylyl cyclase. Biopsies of colonic mucosa from IBS patients released proteases that induced persistent PAR2-dependent hyperexcitability of nociceptors, and PAR2 association with β-arrestins, which mediate endocytosis. Conjugation to cholestanol promoted delivery and retention of antagonists in endosomes containing PAR2. A cholestanol-conjugated PAR2 antagonist prevented persistent trypsin- and IBS protease-induced hyperexcitability of nociceptors. The results reveal that PAR2 signaling from endosomes underlies the persistent hyperexcitability of nociceptors that mediates chronic pain of IBS. Endosomally targeted PAR2 antagonists are potential therapies for IBS pain. GPCRs in endosomes transmit signals that contribute to human diseases.",
keywords = "Endosomes, Pain, Proteases, Receptors",
author = "Jimenez-Vargas, {Nestor N.} and Pattison, {Luke A.} and Peishen Zhao and TinaMarie Lieu and Rocco Latorre and Jensen, {Dane D.} and Joel Castro and Luigi Aurelio and Le, {Giang T.} and Bernard Flynn and Herenbrink, {Carmen Klein} and Yeatman, {Holly R.} and Laura Edgington-Mitchell and Porter, {Christopher J.H.} and Halls, {Michelle L.} and Meritxell Canals and Veldhuis, {Nicholas A.} and Poole, {Daniel P.} and Peter McLean and Hicks, {Gareth A.} and Nicole Scheff and Elyssa Chen and Aditi Bhattacharya and Schmidt, {Brian L.} and Brierley, {Stuart M.} and Vanner, {Stephen J.} and Bunnett, {Nigel W.}",
year = "2018",
month = "7",
day = "31",
doi = "10.1073/pnas.1721891115",
language = "English",
volume = "115",
pages = "E7438--E7447",
journal = "Proceedings of the National Academy of Sciences",
issn = "0027-8424",
publisher = "National Acad Sciences",
number = "31",

}

Jimenez-Vargas, NN, Pattison, LA, Zhao, P, Lieu, T, Latorre, R, Jensen, DD, Castro, J, Aurelio, L, Le, GT, Flynn, B, Herenbrink, CK, Yeatman, HR, Edgington-Mitchell, L, Porter, CJH, Halls, ML, Canals, M, Veldhuis, NA, Poole, DP, McLean, P, Hicks, GA, Scheff, N, Chen, E, Bhattacharya, A, Schmidt, BL, Brierley, SM, Vanner, SJ & Bunnett, NW 2018, 'Protease-activated receptor-2 in endosomes signals persistent pain of irritable bowel syndrome' Proceedings of the National Academy of Sciences of the United States of America, vol. 115, no. 31, pp. E7438-E7447. https://doi.org/10.1073/pnas.1721891115

Protease-activated receptor-2 in endosomes signals persistent pain of irritable bowel syndrome. / Jimenez-Vargas, Nestor N.; Pattison, Luke A.; Zhao, Peishen; Lieu, TinaMarie; Latorre, Rocco; Jensen, Dane D.; Castro, Joel; Aurelio, Luigi; Le, Giang T.; Flynn, Bernard; Herenbrink, Carmen Klein; Yeatman, Holly R.; Edgington-Mitchell, Laura; Porter, Christopher J.H.; Halls, Michelle L.; Canals, Meritxell; Veldhuis, Nicholas A.; Poole, Daniel P.; McLean, Peter; Hicks, Gareth A.; Scheff, Nicole; Chen, Elyssa; Bhattacharya, Aditi; Schmidt, Brian L.; Brierley, Stuart M.; Vanner, Stephen J.; Bunnett, Nigel W.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 115, No. 31, 31.07.2018, p. E7438-E7447.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Protease-activated receptor-2 in endosomes signals persistent pain of irritable bowel syndrome

AU - Jimenez-Vargas, Nestor N.

AU - Pattison, Luke A.

AU - Zhao, Peishen

AU - Lieu, TinaMarie

AU - Latorre, Rocco

AU - Jensen, Dane D.

AU - Castro, Joel

AU - Aurelio, Luigi

AU - Le, Giang T.

AU - Flynn, Bernard

AU - Herenbrink, Carmen Klein

AU - Yeatman, Holly R.

AU - Edgington-Mitchell, Laura

AU - Porter, Christopher J.H.

AU - Halls, Michelle L.

AU - Canals, Meritxell

AU - Veldhuis, Nicholas A.

AU - Poole, Daniel P.

AU - McLean, Peter

AU - Hicks, Gareth A.

AU - Scheff, Nicole

AU - Chen, Elyssa

AU - Bhattacharya, Aditi

AU - Schmidt, Brian L.

AU - Brierley, Stuart M.

AU - Vanner, Stephen J.

AU - Bunnett, Nigel W.

PY - 2018/7/31

Y1 - 2018/7/31

N2 - Once activated at the surface of cells, G protein-coupled receptors (GPCRs) redistribute to endosomes, where they can continue to signal. Whether GPCRs in endosomes generate signals that contribute to human disease is unknown. We evaluated endosomal signaling of protease-activated receptor-2 (PAR2), which has been proposed to mediate pain in patients with irritable bowel syndrome (IBS). Trypsin, elastase, and cathepsin S, which are activated in the colonic mucosa of patients with IBS and in experimental animals with colitis, caused persistent PAR2-dependent hyperexcitability of nociceptors, sensitization of colonic afferent neurons to mechanical stimuli, and somatic mechanical allodynia. Inhibitors of clathrin- and dynamin-dependent endocytosis and of mitogen-activated protein kinase kinase-1 prevented trypsin-induced hyperexcitability, sensitization, and allodynia. However, they did not affect elastase- or cathepsin S-induced hyperexcitability, sensitization, or allodynia. Trypsin stimulated endocytosis of PAR2, which signaled from endosomes to activate extracellular signal-regulated kinase. Elastase and cathepsin S did not stimulate endocytosis of PAR2, which signaled from the plasma membrane to activate adenylyl cyclase. Biopsies of colonic mucosa from IBS patients released proteases that induced persistent PAR2-dependent hyperexcitability of nociceptors, and PAR2 association with β-arrestins, which mediate endocytosis. Conjugation to cholestanol promoted delivery and retention of antagonists in endosomes containing PAR2. A cholestanol-conjugated PAR2 antagonist prevented persistent trypsin- and IBS protease-induced hyperexcitability of nociceptors. The results reveal that PAR2 signaling from endosomes underlies the persistent hyperexcitability of nociceptors that mediates chronic pain of IBS. Endosomally targeted PAR2 antagonists are potential therapies for IBS pain. GPCRs in endosomes transmit signals that contribute to human diseases.

AB - Once activated at the surface of cells, G protein-coupled receptors (GPCRs) redistribute to endosomes, where they can continue to signal. Whether GPCRs in endosomes generate signals that contribute to human disease is unknown. We evaluated endosomal signaling of protease-activated receptor-2 (PAR2), which has been proposed to mediate pain in patients with irritable bowel syndrome (IBS). Trypsin, elastase, and cathepsin S, which are activated in the colonic mucosa of patients with IBS and in experimental animals with colitis, caused persistent PAR2-dependent hyperexcitability of nociceptors, sensitization of colonic afferent neurons to mechanical stimuli, and somatic mechanical allodynia. Inhibitors of clathrin- and dynamin-dependent endocytosis and of mitogen-activated protein kinase kinase-1 prevented trypsin-induced hyperexcitability, sensitization, and allodynia. However, they did not affect elastase- or cathepsin S-induced hyperexcitability, sensitization, or allodynia. Trypsin stimulated endocytosis of PAR2, which signaled from endosomes to activate extracellular signal-regulated kinase. Elastase and cathepsin S did not stimulate endocytosis of PAR2, which signaled from the plasma membrane to activate adenylyl cyclase. Biopsies of colonic mucosa from IBS patients released proteases that induced persistent PAR2-dependent hyperexcitability of nociceptors, and PAR2 association with β-arrestins, which mediate endocytosis. Conjugation to cholestanol promoted delivery and retention of antagonists in endosomes containing PAR2. A cholestanol-conjugated PAR2 antagonist prevented persistent trypsin- and IBS protease-induced hyperexcitability of nociceptors. The results reveal that PAR2 signaling from endosomes underlies the persistent hyperexcitability of nociceptors that mediates chronic pain of IBS. Endosomally targeted PAR2 antagonists are potential therapies for IBS pain. GPCRs in endosomes transmit signals that contribute to human diseases.

KW - Endosomes

KW - Pain

KW - Proteases

KW - Receptors

UR - http://www.scopus.com/inward/record.url?scp=85051717936&partnerID=8YFLogxK

U2 - 10.1073/pnas.1721891115

DO - 10.1073/pnas.1721891115

M3 - Article

VL - 115

SP - E7438-E7447

JO - Proceedings of the National Academy of Sciences

JF - Proceedings of the National Academy of Sciences

SN - 0027-8424

IS - 31

ER -