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

Jimenez-Vargas, Nestor N.; Pattison, Luke A.; Zhao, Peishen; Lieu, Tina Marie; 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.

Abstract

© 2018 National Academy of Sciences. All rights reserved. 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.

Citation

Jimenez-Vargas, N. N., Pattison, L. A., Zhao, P., Lieu, T. M., Latorre, R., Jensen, D. D., …Bunnett, N. W. (2018). Protease-activated receptor-2 in endosomes signals persistent pain of irritable bowel syndrome. Proceedings of the National Academy of Sciences, 115(31), E7438-E7447. https://doi.org/10.1073/pnas.1721891115

• Journal Article Type: Article
• Acceptance Date: Oct 6, 2017
• Online Publication Date: Jul 16, 2018
• Publication Date: Jul 31, 2018
• Deposit Date: Jan 13, 2020
• Journal: Proceedings of the National Academy of Sciences
• Print ISSN: 0027-8424
• Electronic ISSN: 1091-6490
• Publisher: National Academy of Sciences
• Peer Reviewed: Peer Reviewed
• Volume: 115
• Issue: 31
• Pages: E7438-E7447
• DOI: https://doi.org/10.1073/pnas.1721891115
• Keywords: Endosomes; Pain; Proteases; Receptors; beta arrestin; cathepsin S; cholestanol; elastase; enzyme inhibitor; mitogen activated protein kinase kinase 1; proteinase activated receptor 2; trypsin; mitogen activated protein kinase; proteinase activated recepto
• Public URL: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85051717936&doi=10.1073%2fpnas.1721891115&partnerID=40&md5=914c8262c51b6d590202f02407da0303
• Publisher URL: https://www.pnas.org/content/115/31/E7438