Neurokinin 1 receptor signaling in endosomes mediates sustained nociception and is a viable therapeutic target for prolonged pain relief

Dane D Jensen, TinaMarie Lieu, Michelle L Halls, Nicholas A Veldhuis, Wendy L Imlach, Quynh N Mai, Daniel P Poole, Tim Quach, Luigi Aurelio, Joshus Conner, Carmen Klein Herenbrink, Nicholas Barlow, Jamie S Simpson, Martin J Scanlon, Bimbil Graham, Adam McCluskey, Phillip J Robinson, Virginie Escriou, Romina Nassini, Serena Materazzi & 6 others Pierangelo Geppetti, Gareth A Hicks, Macdonald J Christie, Christopher J H Porter, Meritxell Canals, Nigel W Bunnett

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Typically considered to be cell surface sensors of extracellular signals, heterotrimeric GTP-binding protein (G protein)-coupled receptors (GPCRs) control many pathophysiological processes and are the target of 30% of therapeutic drugs. Activated receptors redistribute to endosomes, but researchers have yet to explore whether endosomal receptors generate signals that control complex processes in vivo and are viable therapeutic targets. We report that the substance P (SP) neurokinin 1 receptor (NK1R) signals from endosomes to induce sustained excitation of spinal neurons and pain transmission and that specific antagonism of the NK1R in endosomes with membrane-anchored drug conjugates provides more effective and sustained pain relief than conventional plasma membrane-targeted antagonists. Pharmacological and genetic disruption of clathrin, dynamin, and b-arrestin blocked SP-induced NK1R endocytosis and prevented SP-stimulated activation of cytosolic protein kinase C and nuclear extracellular signal-regulated kinase, as well as transcription. Endocytosis inhibitors prevented sustained SP-induced excitation of neurons in spinal cord slices in vitro and attenuated nociception in vivo. When conjugated to cholestanol to promote endosomal targeting, NK1R antagonists selectively inhibited endosomal signaling and sustained neuronal excitation. Cholestanol conjugation amplified and prolonged the antinociceptive actions of NK1R antagonists. These results reveal a critical role for endosomal signaling of the NK1R in the complex pathophysiology of pain and demonstrate the use of endosomally targeted GPCR antagonists.

Original languageEnglish
Article numberaal3447
Number of pages15
JournalScience Translational Medicine
Volume9
Issue number392
DOIs
Publication statusPublished - 31 May 2017

Cite this

Jensen, Dane D ; Lieu, TinaMarie ; Halls, Michelle L ; Veldhuis, Nicholas A ; Imlach, Wendy L ; Mai, Quynh N ; Poole, Daniel P ; Quach, Tim ; Aurelio, Luigi ; Conner, Joshus ; Klein Herenbrink, Carmen ; Barlow, Nicholas ; Simpson, Jamie S ; Scanlon, Martin J ; Graham, Bimbil ; McCluskey, Adam ; Robinson, Phillip J ; Escriou, Virginie ; Nassini, Romina ; Materazzi, Serena ; Geppetti, Pierangelo ; Hicks, Gareth A ; Christie, Macdonald J ; Porter, Christopher J H ; Canals, Meritxell ; Bunnett, Nigel W. / Neurokinin 1 receptor signaling in endosomes mediates sustained nociception and is a viable therapeutic target for prolonged pain relief. In: Science Translational Medicine. 2017 ; Vol. 9, No. 392.
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title = "Neurokinin 1 receptor signaling in endosomes mediates sustained nociception and is a viable therapeutic target for prolonged pain relief",
abstract = "Typically considered to be cell surface sensors of extracellular signals, heterotrimeric GTP-binding protein (G protein)-coupled receptors (GPCRs) control many pathophysiological processes and are the target of 30{\%} of therapeutic drugs. Activated receptors redistribute to endosomes, but researchers have yet to explore whether endosomal receptors generate signals that control complex processes in vivo and are viable therapeutic targets. We report that the substance P (SP) neurokinin 1 receptor (NK1R) signals from endosomes to induce sustained excitation of spinal neurons and pain transmission and that specific antagonism of the NK1R in endosomes with membrane-anchored drug conjugates provides more effective and sustained pain relief than conventional plasma membrane-targeted antagonists. Pharmacological and genetic disruption of clathrin, dynamin, and b-arrestin blocked SP-induced NK1R endocytosis and prevented SP-stimulated activation of cytosolic protein kinase C and nuclear extracellular signal-regulated kinase, as well as transcription. Endocytosis inhibitors prevented sustained SP-induced excitation of neurons in spinal cord slices in vitro and attenuated nociception in vivo. When conjugated to cholestanol to promote endosomal targeting, NK1R antagonists selectively inhibited endosomal signaling and sustained neuronal excitation. Cholestanol conjugation amplified and prolonged the antinociceptive actions of NK1R antagonists. These results reveal a critical role for endosomal signaling of the NK1R in the complex pathophysiology of pain and demonstrate the use of endosomally targeted GPCR antagonists.",
author = "Jensen, {Dane D} and TinaMarie Lieu and Halls, {Michelle L} and Veldhuis, {Nicholas A} and Imlach, {Wendy L} and Mai, {Quynh N} and Poole, {Daniel P} and Tim Quach and Luigi Aurelio and Joshus Conner and {Klein Herenbrink}, Carmen and Nicholas Barlow and Simpson, {Jamie S} and Scanlon, {Martin J} and Bimbil Graham and Adam McCluskey and Robinson, {Phillip J} and Virginie Escriou and Romina Nassini and Serena Materazzi and Pierangelo Geppetti and Hicks, {Gareth A} and Christie, {Macdonald J} and Porter, {Christopher J H} and Meritxell Canals and Bunnett, {Nigel W}",
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Jensen, DD, Lieu, T, Halls, ML, Veldhuis, NA, Imlach, WL, Mai, QN, Poole, DP, Quach, T, Aurelio, L, Conner, J, Klein Herenbrink, C, Barlow, N, Simpson, JS, Scanlon, MJ, Graham, B, McCluskey, A, Robinson, PJ, Escriou, V, Nassini, R, Materazzi, S, Geppetti, P, Hicks, GA, Christie, MJ, Porter, CJH, Canals, M & Bunnett, NW 2017, 'Neurokinin 1 receptor signaling in endosomes mediates sustained nociception and is a viable therapeutic target for prolonged pain relief' Science Translational Medicine, vol. 9, no. 392, aal3447. https://doi.org/10.1126/scitranslmed.aal3447

Neurokinin 1 receptor signaling in endosomes mediates sustained nociception and is a viable therapeutic target for prolonged pain relief. / Jensen, Dane D; Lieu, TinaMarie; Halls, Michelle L; Veldhuis, Nicholas A; Imlach, Wendy L; Mai, Quynh N; Poole, Daniel P; Quach, Tim; Aurelio, Luigi; Conner, Joshus; Klein Herenbrink, Carmen; Barlow, Nicholas; Simpson, Jamie S; Scanlon, Martin J; Graham, Bimbil; McCluskey, Adam; Robinson, Phillip J; Escriou, Virginie; Nassini, Romina; Materazzi, Serena; Geppetti, Pierangelo; Hicks, Gareth A; Christie, Macdonald J; Porter, Christopher J H; Canals, Meritxell; Bunnett, Nigel W.

In: Science Translational Medicine, Vol. 9, No. 392, aal3447, 31.05.2017.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Neurokinin 1 receptor signaling in endosomes mediates sustained nociception and is a viable therapeutic target for prolonged pain relief

AU - Jensen, Dane D

AU - Lieu, TinaMarie

AU - Halls, Michelle L

AU - Veldhuis, Nicholas A

AU - Imlach, Wendy L

AU - Mai, Quynh N

AU - Poole, Daniel P

AU - Quach, Tim

AU - Aurelio, Luigi

AU - Conner, Joshus

AU - Klein Herenbrink, Carmen

AU - Barlow, Nicholas

AU - Simpson, Jamie S

AU - Scanlon, Martin J

AU - Graham, Bimbil

AU - McCluskey, Adam

AU - Robinson, Phillip J

AU - Escriou, Virginie

AU - Nassini, Romina

AU - Materazzi, Serena

AU - Geppetti, Pierangelo

AU - Hicks, Gareth A

AU - Christie, Macdonald J

AU - Porter, Christopher J H

AU - Canals, Meritxell

AU - Bunnett, Nigel W

PY - 2017/5/31

Y1 - 2017/5/31

N2 - Typically considered to be cell surface sensors of extracellular signals, heterotrimeric GTP-binding protein (G protein)-coupled receptors (GPCRs) control many pathophysiological processes and are the target of 30% of therapeutic drugs. Activated receptors redistribute to endosomes, but researchers have yet to explore whether endosomal receptors generate signals that control complex processes in vivo and are viable therapeutic targets. We report that the substance P (SP) neurokinin 1 receptor (NK1R) signals from endosomes to induce sustained excitation of spinal neurons and pain transmission and that specific antagonism of the NK1R in endosomes with membrane-anchored drug conjugates provides more effective and sustained pain relief than conventional plasma membrane-targeted antagonists. Pharmacological and genetic disruption of clathrin, dynamin, and b-arrestin blocked SP-induced NK1R endocytosis and prevented SP-stimulated activation of cytosolic protein kinase C and nuclear extracellular signal-regulated kinase, as well as transcription. Endocytosis inhibitors prevented sustained SP-induced excitation of neurons in spinal cord slices in vitro and attenuated nociception in vivo. When conjugated to cholestanol to promote endosomal targeting, NK1R antagonists selectively inhibited endosomal signaling and sustained neuronal excitation. Cholestanol conjugation amplified and prolonged the antinociceptive actions of NK1R antagonists. These results reveal a critical role for endosomal signaling of the NK1R in the complex pathophysiology of pain and demonstrate the use of endosomally targeted GPCR antagonists.

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DO - 10.1126/scitranslmed.aal3447

M3 - Article

VL - 9

JO - Science Translational Medicine

JF - Science Translational Medicine

SN - 1946-6234

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M1 - aal3447

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