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The effect of μ-opioid receptor polymorphisms on receptor signalling systems

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posted on 29.03.2022, 00:18 by Alisa Knapman
There is significant variation in individual response to opioid drugs, one cause of which is likely to be polymorphisms on the opioid receptors themselves. The μ-opioid receptor (MOPr) is the primary site of action for most analgesic opioids. Previous studies have identified a number of naturally occurring single nucleotide polymorphisms (SNPs) in the coding region of MOPr. The A118G SNP (N40D), present at allelic frequencies ranging from 10 – 50%, has been associated with diverse phenotypic effects as well as differences in receptor signalling in vitro, with little consistency between studies. Few studies have examined the consequences of other MOPr polymorphisms on receptor function, or potential ligand and pathway specific effects. In this study, the relative potency and efficacy of a range of clinically important and/or structurally distinct opioid ligands were assessed in Chinese hamster ovary (CHO) cells and mouse pituitary neuroblastoma (AtT-20) cells stably transfected with human wild type MOPr and 5 naturally occurring MOPr variants, N40D, A6V, L85I, R260H and R265H. MOPr surface expression levels were similar for all variants examined. MOPr activation was measured using a membrane-potential assay of adenylyl cyclase (AC) inhibition, a whole-cell ELISA of extracellular signal-regulated kinase 1/2 (ERK1/2) phosphorylation, and a membrane-potential assay of G protein-activated potassium channels (GIRKs). In cells expressing MOPr-N40D, buprenorphine inhibition of AC and stimulation of ERK1/2 was significantly reduced, with GIRK activation unaffected. In cells expressing MOPR-C17T, buprenorphine signalling was abolished, with a loss of potency of morphine and other ligands. AC inhibition via non-morphinan opioids was enhanced at L85I, while a significant loss of potency for many opioids was observed at R260H. There were minor alterations in the signalling profile of R265H. These results suggest that MOPr SNPs have the potential to significantly alter receptor function, and may contribute to the individual variability in response to opioid analgesics observed clinically.

History

Alternative Title

Effect of mu-opioid receptor polymorphisms on receptor signalling systems

Table of Contents

Chapter 1. Introduction -- Chapter 2. Cellular signalling of non-synonomous single nucleotide polymorphisms of the human μ-opioid receptor -- Chapter 3. A real-time, fluorescence-based assay for measuring μ-opioid receptor modulation of adenylyl cyclase activity in Chinese hamster ovary cells -- Chapter 4. A continuous, fluorescence-based assay of μ-opioid receptor activation in AtT-20 cells -- Chapter 5. Buprenorphine signalling is compromised at the N40D polymorphism of the human μ-opioid receptor in vitro -- Chapter 6. The A6V polymorphism of the human μ-opioid receptor negatively impacts signalling of morphine and endogenous opioids in vitro -- Chapter 7. Mu-opioid receptor polymorphisms differentially affect receptor signalling via adenylyl cyclase inhibition and ERK1/2 phosphorylation -- Chaper 8. General discussion and summary.

Notes

Includes bibliographic references Thesis by publication. Spine title: The effect of mu-opioid receptor polymorphisms on receptor signalling systems

Awarding Institution

Macquarie University

Degree Type

Thesis PhD

Degree

PhD, Macquarie University, Faculty of Medicine and Health Sciences, Australian School of Advanced Medicine

Department, Centre or School

Australian School of Advanced Medicine

Year of Award

2014

Principal Supervisor

Mark Connor

Rights

Copyright Alisa Knapman 2014. Copyright disclaimer: http://www.copyright.mq.edu.au

Language

English

Extent

1 online resource (xiv, 343 pages) graphs, tables

Former Identifiers

mq:44918 http://hdl.handle.net/1959.14/1073216