Projects per year
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 language | English |
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Article number | aal3447 |
Number of pages | 15 |
Journal | Science Translational Medicine |
Volume | 9 |
Issue number | 392 |
DOIs | |
Publication status | Published - 31 May 2017 |
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ARC Centre of Excellence in Convergent Bio-Nano Science and Technology
Davis, T. (Primary Chief Investigator (PCI)), Boyd, B. (Chief Investigator (CI)), Bunnett, N. (Chief Investigator (CI)), Porter, C. (Chief Investigator (CI)), Caruso, F. (Chief Investigator (CI)), Kent, S. (Chief Investigator (CI)), Thordarson, P. (Chief Investigator (CI)), Kearnes, M. (Chief Investigator (CI)), Gooding, J. (Chief Investigator (CI)), Kavallaris, M. (Chief Investigator (CI)), Thurecht, K. J. (Chief Investigator (CI)), Whittaker, A. K. (Chief Investigator (CI)), Parton, R. (Chief Investigator (CI)), Corrie, S. R. (Chief Investigator (CI)), Johnston, A. (Chief Investigator (CI)), McGhee, J. (Chief Investigator (CI)), Greguric, I. D. (Partner Investigator (PI)), Stevens, M. M. (Partner Investigator (PI)), Lewis, J. S. (Partner Investigator (PI)), Lee, D. S. (Partner Investigator (PI)), Alexander, C. (Partner Investigator (PI)), Dawson, K. (Partner Investigator (PI)), Hawker, C. (Partner Investigator (PI)), Haddleton, D. (Partner Investigator (PI)), Thierry, B. (Chief Investigator (CI)), Prestidge, C. A. (Chief Investigator (CI)), Meyer, A. (Project Manager), Jones-Jayasinghe, N. (Project Manager), Voelcker, N. (Chief Investigator (CI)), Nann, T. (Chief Investigator (CI)) & McLean, K. (Partner Investigator (PI))
Australian Research Council (ARC), Monash University, University of Melbourne, University of New South Wales (UNSW), University of Queensland , University of South Australia, Monash University – Internal Faculty Contribution, University of Wisconsin Madison, Memorial Sloan Kettering Cancer Center, University of California System, University College Dublin, Imperial College London, University of Warwick, Sungkyunkwan University, Australian Nuclear Science and Technology Organisation (ANSTO) , University of Nottingham
30/06/14 → 29/06/21
Project: Research
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Signalosomes and compartmentalisation in cellular homeostasis and disease
Halls, M. (Primary Chief Investigator (PCI))
National Health and Medical Research Council (NHMRC) (Australia)
1/01/14 → 31/10/19
Project: Research
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Mechanisms of protease-driven visceral pain
Graham, B. (Primary Chief Investigator (PCI)), Brierley, S. (Chief Investigator (CI)), Poole, D. (Chief Investigator (CI)) & Bunnett, N. (Associate Investigator (AI))
National Health and Medical Research Council (NHMRC) (Australia)
1/01/13 → 31/12/15
Project: Research