Targeted inhibition of mu-opioid receptors in neuronal subpopulations by membrane-tethered Naloxo-DART antagonists

J. Sanchez; A. Bonifazi; S. Groom; M. O. Sambrook; G. A. Camacho-Hernandez; E. J. Kuijer; S. Karimi; K. U. Therese; M. Hauge Pedersen; C. Rauffenbart; M. Canals; A. H. Newman; C. P. Bailey; J. A. Javitch; J. R. Lane

doi:10.1016/j.chembiol.2025.11.002

Targeted inhibition of mu-opioid receptors in neuronal subpopulations by membrane-tethered Naloxo-DART antagonists

J. Sanchez, A. Bonifazi, S. Groom, M. O. Sambrook, G. A. Camacho-Hernandez, E. J. Kuijer, S. Karimi, K. U. Therese, M. Hauge Pedersen, C. Rauffenbart, M. Canals, A. H. Newman, C. P. Bailey, J. A. Javitch, J. R. Lane

Research output: Contribution to journal › Article › peer-review

1 Citation (SciVal)

Abstract

Opioid receptors are expressed in virtually all neural loci contributing to the experience of pain. Due to this widespread expression, the contribution of specific cell types to the analgesic properties and deleterious side effects of opioids remains incompletely understood. Linking the activity of specific receptors in defined cells to behavioral or physiological processes remains a major challenge of translational pharmacology. In this study, we describe the development of drugs acutely restricted by membrane tethering (DART) antagonists that contain an antagonist naloxone moiety linked to a Halo-tag reactive group. The optimized Naloxo-DART displayed robust blockade of a MOR agonist only when cells co-expressed a Halo-tagged membrane tether. We use the Naloxo-DART delivered in vivo to selectively block MORs in locus coeruleus neurons in brain slide preparations. The Naloxo-DART provides a powerful approach for elucidating the physiological roles of MORs expressed in specific neuronal populations with acute spatiotemporal control.

Original language	English
Pages (from-to)	1473-1485.e9
Number of pages	23
Journal	Cell Chemical Biology
Volume	32
Issue number	12
Early online date	3 Dec 2025
DOIs	https://doi.org/10.1016/j.chembiol.2025.11.002
Publication status	Published - 18 Dec 2025

Data Availability Statement

• All experimental data shown in data figures and supplemental figures
are provided in Data S2.
• This paper does not report original code.
• Any additional information required to reanalyze the data reported in this
paper is available from the lead contact upon request.

Acknowledgements

We thank Jackie Glenn (University of Nottingham) for excellent technical support
and the School of Life Sciences Imaging (SLIM, University of Nottingham) for
support with imaging experiments.

Funding

BBSRC grant: BB/T01363X/1 Funding for this research was provided by the Biotechnology and Biological Sciences Research Council BB/T013966/1 (J.R.L., M.C., and C.P.B.), the NIDA Intramural Research Program Z1A DA000424 (A.H.N.), a Schaefer Research Scholars Program Award (Columbia University, New York, USA) (J.R.L. and J.A.J) and the Hope for Depression Research Foundation (J.A.J).

Keywords

chemogenetics
DART
drugs acutely restricted by membrane tethering
G-protein-coupled receptor
opioid receptor

ASJC Scopus subject areas

Biochemistry
Molecular Medicine
Molecular Biology
Pharmacology
Drug Discovery
Clinical Biochemistry

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Sanchez, J., Bonifazi, A., Groom, S., Sambrook, M. O., Camacho-Hernandez, G. A., Kuijer, E. J., Karimi, S., Therese, K. U., Hauge Pedersen, M., Rauffenbart, C., Canals, M., Newman, A. H., Bailey, C. P., Javitch, J. A., & Lane, J. R. (2025). Targeted inhibition of mu-opioid receptors in neuronal subpopulations by membrane-tethered Naloxo-DART antagonists. Cell Chemical Biology, 32(12), 1473-1485.e9. https://doi.org/10.1016/j.chembiol.2025.11.002

Sanchez, J, Bonifazi, A, Groom, S, Sambrook, MO, Camacho-Hernandez, GA, Kuijer, EJ, Karimi, S, Therese, KU, Hauge Pedersen, M, Rauffenbart, C, Canals, M, Newman, AH, Bailey, CP, Javitch, JA & Lane, JR 2025, 'Targeted inhibition of mu-opioid receptors in neuronal subpopulations by membrane-tethered Naloxo-DART antagonists', Cell Chemical Biology, vol. 32, no. 12, pp. 1473-1485.e9. https://doi.org/10.1016/j.chembiol.2025.11.002

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abstract = "Opioid receptors are expressed in virtually all neural loci contributing to the experience of pain. Due to this widespread expression, the contribution of specific cell types to the analgesic properties and deleterious side effects of opioids remains incompletely understood. Linking the activity of specific receptors in defined cells to behavioral or physiological processes remains a major challenge of translational pharmacology. In this study, we describe the development of drugs acutely restricted by membrane tethering (DART) antagonists that contain an antagonist naloxone moiety linked to a Halo-tag reactive group. The optimized Naloxo-DART displayed robust blockade of a MOR agonist only when cells co-expressed a Halo-tagged membrane tether. We use the Naloxo-DART delivered in vivo to selectively block MORs in locus coeruleus neurons in brain slide preparations. The Naloxo-DART provides a powerful approach for elucidating the physiological roles of MORs expressed in specific neuronal populations with acute spatiotemporal control.",

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AU - Hauge Pedersen, M.

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Targeted inhibition of mu-opioid receptors in neuronal subpopulations by membrane-tethered Naloxo-DART antagonists

Abstract

Data Availability Statement

Acknowledgements

Funding

Keywords

ASJC Scopus subject areas

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