Cannabinoid tolerance relies on CB1 receptor ubiquitination by NEDD4L
Abstract
Cannabinoids, the active components of cannabis, lose efficacy after repeated use due to tolerance, limiting their therapeutic value and safety. Although short-term desensitization of cannabinoid receptors is well understood, the molecular basis of long-term cannabinoid tolerance has remained unclear. This study identifies a molecular mechanism by which cannabinoids trigger selective ubiquitination and proteasomal degradation of the CB 1 receptor in the brain. This process is driven by protein kinase C-dependent activation of the E3 ubiquitin ligase neural precursor cell-expressed developmentally downregulated 4-like and is required for behavioral cannabinoid tolerance in mice. These findings provide a molecular framework for cannabinoid tolerance, a process that can contribute to cannabis use disorder and cannabis hyperemesis syndrome, and suggest strategies to improve cannabinoid-based therapies. Cannabinoids, the active components of cannabis, exert numerous acute effects in the brain by engaging cannabinoid CB 1 receptors (CB 1 Rs). However, tolerance emerges rapidly after repeated drug exposure, undermining the efficacy of cannabinoid-based therapies and contributing to cannabis-associated adverse effects. Although the processes of CB 1 R short-t erm desensitization (i.e., receptor uncoupling and internalization) are well characterized, the mechanisms underlying CB 1 R long-term tolerance (i.e., downregulation of receptor protein levels) remain elusive. Here, we identify a ubiquitin-dependent pathway that couples CB 1 R activation to its proteasomal degradation. We show that cannabinoids engage a G q/11 -PLC-P KC signaling cascade that phosphorylates and activates the E3 ubiquitin ligase neural precursor cell-expressed developmentally downregulated 4-like (NEDD4L), promoting its recruitment to CB 1 R and the ubiquitination of four specific lysine residues. This modification targets the receptor for proteasomal clearance, reducing neuronal CB 1 R abundance in vitr o and in the mouse brain. Using molecular, pharmacological, and circuit-s pecific rescue approaches, we demonstrate that preventing CB 1 R ubiquitination stabilizes receptor levels and abolishes behavioral cannabinoid tolerance in mice without impairing acute drug responses. These findings reveal a molecular mechanism that controls CB 1 R stability and identify NEDD4L-mediated ubiquitination as a central driver of cannabinoid tolerance.
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BibTeX
@article{alvaroblazquez2026cannabinoid,
title = {{Cannabinoid tolerance relies on CB1 receptor ubiquitination by NEDD4L}},
author = {Alicia Álvaro-Blázquez and Rui S Rodrigues and Carlos Montero-Fernández and Marta Isasa and Astrid Cannich and Doriane Gisquet and Ignacio Rodríguez-Crespo and Luigi Bellocchio and Giovanni Marsicano and Carlos Costas-Insua and Manuel Guzmán},
journal = {Proceedings of the National Academy of Sciences of the United States of America},
year = {2026},
month = nov,
doi = {10.1073/pnas.2606671123},
url = {https://hal.science/hal-05848884v1},
note = {CC BY-NC-ND 4.0},
}
RIS
TY - JOUR
TI - Cannabinoid tolerance relies on CB1 receptor ubiquitination by NEDD4L
AU - Alicia Álvaro-Blázquez
AU - Rui S Rodrigues
AU - Carlos Montero-Fernández
AU - Marta Isasa
AU - Astrid Cannich
AU - Doriane Gisquet
AU - Ignacio Rodríguez-Crespo
AU - Luigi Bellocchio
AU - Giovanni Marsicano
AU - Carlos Costas-Insua
AU - Manuel Guzmán
PY - 2026
DA - 2026/11/02
JO - Proceedings of the National Academy of Sciences of the United States of America
DO - 10.1073/pnas.2606671123
UR - https://hal.science/hal-05848884v1
AB - Cannabinoids, the active components of cannabis, lose efficacy after repeated use due to tolerance, limiting their therapeutic value and safety. Although short-term desensitization of cannabinoid receptors is well understood, the molecular basis of long-term cannabinoid tolerance has remained unclear. This study identifies a molecular mechanism by which cannabinoids trigger selective ubiquitination and proteasomal degradation of the CB 1 receptor in the brain. This process is driven by protein kinase C-dependent activation of the E3 ubiquitin ligase neural precursor cell-expressed developmentally downregulated 4-like and is required for behavioral cannabinoid tolerance in mice. These findings provide a molecular framework for cannabinoid tolerance, a process that can contribute to cannabis use disorder and cannabis hyperemesis syndrome, and suggest strategies to improve cannabinoid-based therapies. Cannabinoids, the active components of cannabis, exert numerous acute effects in the brain by engaging cannabinoid CB 1 receptors (CB 1 Rs). However, tolerance emerges rapidly after repeated drug exposure, undermining the efficacy of cannabinoid-based therapies and contributing to cannabis-associated adverse effects. Although the processes of CB 1 R short-t erm desensitization (i.e., receptor uncoupling and internalization) are well characterized, the mechanisms underlying CB 1 R long-term tolerance (i.e., downregulation of receptor protein levels) remain elusive. Here, we identify a ubiquitin-dependent pathway that couples CB 1 R activation to its proteasomal degradation. We show that cannabinoids engage a G q/11 -PLC-P KC signaling cascade that phosphorylates and activates the E3 ubiquitin ligase neural precursor cell-expressed developmentally downregulated 4-like (NEDD4L), promoting its recruitment to CB 1 R and the ubiquitination of four specific lysine residues. This modification targets the receptor for proteasomal clearance, reducing neuronal CB 1 R abundance in vitr o and in the mouse brain. Using molecular, pharmacological, and circuit-s pecific rescue approaches, we demonstrate that preventing CB 1 R ubiquitination stabilizes receptor levels and abolishes behavioral cannabinoid tolerance in mice without impairing acute drug responses. These findings reveal a molecular mechanism that controls CB 1 R stability and identify NEDD4L-mediated ubiquitination as a central driver of cannabinoid tolerance.
SN - 0027-8424
SN - 1091-6490
ER -