Berman, P. | bioRxiv Science

bioRxiv2025

Circadian-Related Dynamics of the Endocannabinoid System in Male Mouse Brain

Endocannabinoids (eCBs) and related lipids play crucial roles in brain function, including the regulation of circadian rhythms and sleep. To comprehensively map these molecules, we employed liquid chromatography high-resolution tandem mass spectrometry (LC/HRMS/MS) to quantify 78 lipids across 14 families in seven brain areas of male mice at four time points throughout the day (every six hours), and during sleep initiation. We found that most eCBs from the fatty acids (FAs) family, particularly arachidonic acid (AA), were highly abundant in the mouse brain in all brain areas and during the circadian rhythm. High eCBs abundance was shown in deeper brain areas, while temporal differences using the Cosinor analysis revealed 26 eCBs behaving in a circadian rhythm response, with linolenic acid (LnA) being the only lipid to show rhythmicity across all brain areas. Sleep initiation (ZT1) was associated with increased N-acylphosphatidylethanolamine phospholipase D (NAPE-PLD) activity and N-acylethanolamide (NAE) levels in the cortex and hippocampus, while wake extension (WEx) altered 2-monoacylglycerol (2-MAG) metabolism and increased cannabinoid receptor 1 (CB1) expression. These findings provide a detailed lipidomic map of eCBs and related lipids in the male mouse brain, highlighting their area-specific distribution, circadian regulation, and involvement in sleep/wake transitions. Given the link between sleep disruption and neurodegeneration, future studies should investigate whether the observed eCB dysregulation contributes to sleep disturbances in these conditions, and if targeting these pathways offers novel therapeutic strategies. Significant StatementThis comprehensive study provides a high-dimensional map of eCBs and related lipids in the male mouse brain, revealing intricate spatial and temporal dynamics highlighting the role of these lipids in regulating fundamental physiological processes such as circadian rhythm and sleep.

neuroscience↗

bioRxiv2023

Cannabinoid Combination Targets NOTCH1-Mutated T-ALL Through the Integrated Stress Response Pathway

In T-cell acute lymphoblastic leukemia (T-ALL) more than 50% of cases display autoactivation of Notch1 signaling, leading to oncogenic transformation. We have previously identified a specific chemovar of Cannabis that induces apoptosis by preventing Notch1 maturation in leukemia cells. Here, we isolated three cannabinoids from this chemovar that synergistically mimic the effects of the whole extract. Two were previously known, Cannabidiol (CBD) and Cannabidivarin (CBDV); whereas the third cannabinoid, which we termed 331-18A, was identified and fully characterized in this study. We demonstrated that these cannabinoids act through Cannabinoid receptor type 2 and TRPV1 to activate the integrated stress response pathway by depleting intracellular Ca2+. This is followed by increased mRNA and protein expression of ATF4, CHOP and CHAC1, which is hindered by inhibiting the upstream initiation factor eIF2. The increased abundance of CHAC1 prevents Notch1 maturation, thereby reducing the levels of the active Notch1 intracellular domain, and consequently decreasing cell viability and increasing apoptosis. Treatment with the three isolated molecules resulted in reduced tumor size and weight in-vivo and slowed leukemia progression in mice models. Altogether, this study elucidated the mechanism of action of three distinct cannabinoids in modulating the Notch1 pathway, and constitutes an important step in the establishment of a new therapy for treating NOTCH1-mutated diseases and cancers such as T-ALL.

cancer biology↗