Reversal of diet-induced hepatic steatosis by peripheral CB1 receptor blockade in mice is p53/miRNA-22/SIRT1/PPARα dependent

• Published in: Molecular metabolism (Germany) Vol. 42; p. 101087
• Main Authors: Azar, Shahar, Udi, Shiran, Drori, Adi, Hadar, Rivka, Nemirovski, Alina, Vemuri, Kiran V., Miller, Maya, Sherill-Rofe, Dana, Arad, Yhara, Gur-Wahnon, Devorah, Li, Xiaoling, Makriyannis, Alexandros, Ben-Zvi, Danny, Tabach, Yuval, Ben-Dov, Iddo Z., Tam, Joseph
• Format: Journal Article
• Language: English
• Published: Germany Elsevier GmbH 01.12.2020; Elsevier
• Subjects: Endocannabinoids; Fatty liver; microRNAs; Nuclear receptor; Obesity; Original; microRNAs; Obesity; Nuclear receptor; Fatty liver; Endocannabinoids
• ISSN: 2212-8778; 2212-8778
• DOI: 10.1016/j.molmet.2020.101087

The endocannabinoid (eCB) system is increasingly recognized as being crucially important in obesity-related hepatic steatosis. By activating the hepatic cannabinoid-1 receptor (CB1R), eCBs modulate lipogenesis and fatty acid oxidation. However, the underlying molecular mechanisms are largely unknown. We combined unbiased bioinformatics techniques, mouse genetic manipulations, multiple pharmacological, molecular, and cellular biology approaches, and genomic sequencing to systematically decipher the role of the hepatic CB1R in modulating fat utilization in the liver and explored the downstream molecular mechanisms. Using an unbiased normalized phylogenetic profiling analysis, we found that the CB1R evolutionarily coevolves with peroxisome proliferator-activated receptor-alpha (PPARα), a key regulator of hepatic lipid metabolism. In diet-induced obese (DIO) mice, peripheral CB1R blockade (using AM6545) induced the reversal of hepatic steatosis and improved liver injury in WT, but not in PPARα−/− mice. The antisteatotic effect mediated by AM6545 in WT DIO mice was accompanied by increased hepatic expression and activity of PPARα as well as elevated hepatic levels of the PPARα-activating eCB-like molecules oleoylethanolamide and palmitoylethanolamide. Moreover, AM6545 was unable to rescue hepatic steatosis in DIO mice lacking liver sirtuin 1 (SIRT1), an upstream regulator of PPARα. Both of these signaling molecules were modulated by the CB1R as measured in hepatocytes exposed to lipotoxic conditions or treated with CB1R agonists in the absence/presence of AM6545. Furthermore, using microRNA transcriptomic profiling, we found that the CB1R regulated the hepatic expression, acetylation, and transcriptional activity of p53, resulting in the enhanced expression of miR-22, which was found to specifically target SIRT1 and PPARα. We provide strong evidence for a functional role of the p53/miR-22/SIRT1/PPARα signaling pathway in potentially mediating the antisteatotic effect of peripherally restricted CB1R blockade. The elucidated molecular signaling pathway by which hepatic CB1R regulates hepatic lipid homeostasis. Activation of hepatic CB1R increases p53 expression, acetylation, nuclear localization,and transcriptional activity that together promote the expression of miR-22, which in turn reduces the expression and activity of PPARα and SIRT1. These events lead to reduced fatty acid oxidation and fat accumulation in the liver (left). Treatment with AM6545, a peripherally restricted CB1R antagonist, disrupts this signaling pathway and ameliorates hepatic steatosis by also increasing hepatic OEA and PEA, which may activate PPARα directly, resulting in the enhanced efficacy of AM6545 in ameliorating hepatic steatosis (right). [Display omitted] •PPARα is the top-ranked NAFLD-related gene that is coevolved with the CB1R.•Hepatic PPARα and SIRT1 expression and activity are regulated by the CB1R.•Reversal of hepatic steatosis by peripheral CB1R blockade requires intact SIRT1 and PPARα signaling.•Hepatic PPARα and SIRT1 are direct targets of miR-22, whose expression is regulated via CB1R-induced modulation of p53 expression, acetylation, and transcriptional activity.