Controlling opioid receptor functional selectivity by targeting distinct subpockets of the orthosteric site
- Memorial Sloan Kettering Cancer Center, United States
- University of North Carolina, United States
- The Bridge Institute, University of Southern California, United States
- Columbia University, United States
- St. Louis College of Pharmacy and Washington University School of Medicine, United States
- Washington University in St. Louis, United States
- Washington University, United States
- University of Florida, United States
- University of Southern California, United States
Abstract
Controlling receptor functional selectivity profiles for opioid receptors is a promising approach for discovering safer analgesics; however, the structural determinants conferring functional selectivity are not well understood. Here we used crystal structures of opioid receptors, including the recently solved active state kappa opioid complex with MP1104, to rationally design novel mixed mu (MOR) and kappa (KOR) opioid receptor agonists with reduced arrestin signaling. Analysis of structure-activity relationships for new MP1104 analogs points to a region between transmembrane 5 (TM5) and extracellular loop (ECL2) as key for modulation of arrestin recruitment to both MOR and KOR. The lead compounds, MP1207 and MP1208, displayed MOR/KOR Gi-partial agonism with diminished arrestin signaling, showed efficient analgesia with attenuated liabilities, including respiratory depression and conditioned place preference and aversion in mice. The findings validate a novel structure-inspired paradigm for achieving beneficial in vivo profiles for analgesia through different mechanisms that include bias, partial agonism, and dual MOR/KOR agonism.
Data availability
All data generated or analysed during this study are included in the manuscript and supporting files.
Article and author information
Author details
Abdelfattah Faouzi
Jonathan A Javitch
Funding
National Institute on Drug Abuse (DA045884)
- Susruta Majumdar
National Institute on Drug Abuse (DA045657)
- Jonathan A Javitch
National Institute on Drug Abuse (DA046487)
- Susruta Majumdar
National Institute on Alcohol Abuse and Alcoholism (AA026949)
- Susruta Majumdar
National Institute on Drug Abuse (DA038858)
- Vsevolod Katritch
National Institute on Drug Abuse (DA035764)
- Bryan L Roth
- Vsevolod Katritch
National Institute on Drug Abuse (DA007242,DA006241)
- Ying Xian Pan
- Gavril W Pasternak
National Institute on Drug Abuse (DA042888,DA046714)
- Ying Xian Pan
National Institute of Mental Health (MH018870)
- Steven G Grinnell
National Institute of Mental Health (MH112205)
- Jonathan A Javitch
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Ethics
Animal experimentation: All animal studies were preapproved by the Institutional Animal Care and Use Committees of University of Florida in accordance with the 2002 National Institutes of Health Guide for the Care and Use of Laboratory Animals. protocols 201808990 and 202011105.
Reviewing Editor
- Olga Boudker, Weill Cornell Medicine, United States
Version history
- Received: March 1, 2020
- Accepted: February 7, 2021
- Accepted Manuscript published: February 8, 2021 (version 1)
- Version of Record published: February 26, 2021 (version 2)
Copyright
© 2021, Uprety et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
Metrics
-
- 4,966
- Page views
-
- 660
- Downloads
-
- 31
- Citations
Article citation count generated by polling the highest count across the following sources: Crossref, Scopus, PubMed Central.
Download links
A two-part list of links to download the article, or parts of the article, in various formats.
Downloads (link to download the article as PDF)
Open citations (links to open the citations from this article in various online reference manager services)
Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)
Controlling opioid receptor functional selectivity by targeting distinct subpockets of the orthosteric site
eLife 10:e56519.
https://doi.org/10.7554/eLife.56519
Further reading
-
- Biochemistry and Chemical Biology
- Cancer Biology
Identification oncogenes is fundamental to revealing the molecular basis of cancer. Here, we found that FOXP2 is overexpressed in human prostate cancer cells and prostate tumors, but its expression is absent in normal prostate epithelial cells and low in benign prostatic hyperplasia. FOXP2 is a FOX transcription factor family member and tightly associated with vocal development. To date, little is known regarding the link of FOXP2 to prostate cancer. We observed that high FOXP2 expression and frequent amplification are significantly associated with high Gleason score. Ectopic expression of FOXP2 induces malignant transformation of mouse NIH3T3 fibroblasts and human prostate epithelial cell RWPE-1. Conversely, FOXP2 knockdown suppresses the proliferation of prostate cancer cells. Transgenic overexpression of FOXP2 in the mouse prostate causes prostatic intraepithelial neoplasia. Overexpression of FOXP2 aberrantly activates oncogenic MET signaling and inhibition of MET signaling effectively reverts the FOXP2-induced oncogenic phenotype. CUT&Tag assay identified FOXP2-binding sites located in MET and its associated gene HGF. Additionally, the novel recurrent FOXP2-CPED1 fusion identified in prostate tumors results in high expression of truncated FOXP2, which exhibit a similar capacity for malignant transformation. Together, our data indicate that FOXP2 is involved in tumorigenicity of prostate.
-
- Biochemistry and Chemical Biology
- Cell Biology
Pancreatic a-cells secrete glucagon, an insulin counter-regulatory peptide hormone critical for the maintenance of glucose homeostasis. Investigation of the function of human a-cells remains a challenge due to the lack of cost-effective purification methods to isolate high-quality a-cells from islets. Here, we use the reaction-based probe diacetylated Zinpyr1 (DA-ZP1) to introduce a novel and simple method for enriching live a-cells from dissociated human islet cells with ~ 95% purity. The a-cells, confirmed by sorting and immunostaining for glucagon, were cultured up to 10 days to form a-pseudoislets. The a-pseudoislets could be maintained in culture without significant loss of viability, and responded to glucose challenge by secreting appropriate levels of glucagon. RNA-sequencing analyses (RNA-seq) revealed that expression levels of key a-cell identity genes were sustained in culture while some of the genes such as DLK1, GSN, SMIM24 were altered in a-pseudoislets in a time-dependent manner. In conclusion, we report a method to sort human primary a-cells with high purity that can be used for downstream analyses such as functional and transcriptional studies.
