M6A methylation-induced NR1D1 ablation disrupts the HSC circadian clock and promotes hepatic fibrosis. (March 2023)

• Record Type: Journal Article
• Authors: Chen, Li; Xia, Siwei; Wang, Feixia; Zhou, Yuanyuan; Wang, Shuqi; Yang, Ting; Li, Yang; Xu, Min; Zhou, Ya; Kong, Desong; Zhang, Zili; Shao, Jiangjuan; Xu, Xuefen; Zhang, Feng; Zheng, Shizhong
• Is Part Of: Pharmacological research. Volume 189(2023)
• Publication Date: 2023-03
• Subjects: AAV9 adeno-associated virus vector serotype 9 -- ALT alanine aminotransferase -- AST aspartate aminotransferase -- ALP Alkaline Phosphatase -- COL1 collagen type I alpha 1 -- CCl4 carbon tetrachloride -- ECM extracellular matrix -- HSC hepatic stellate cell -- HA Hyaluronan -- HA Hyaluronic acid -- IHC immunohistochemistry -- IV-C type IV collagen -- KD knockdown -- KCs Kupffer cells -- LSEC liver sinusoidal endothelial cell -- LDH lactate dehydrogenase -- LN laminin -- OE over expression -- PC-III type III collagen -- qPCR quantitative real-time PCR -- TBIL total bilirubin -- ZT Zeitgeber time -- α-SMA α-smooth muscle actin; Liver fibrosis -- Circadian clock -- Primary hepatic stellate cell -- NR1D1 -- M6A -- Mitochondrial dynamics; Pharmacology -- Periodicals; Pharmacology -- Periodicals; Research -- Periodicals; Médicaments -- Recherche -- Périodiques; Pharmacologie -- Périodiques; 615.105
• DOI: 10.1016/j.phrs.2023.106704
• Languages: English
• ISSNs: 1043-6618

Abstract

Abstract: The roles of nuclear receptor subfamily 1 group d member 1 (NR1D1) and the circadian clock in liver fibrosis remain unclear. Here, we showed that liver clock genes, especially NR1D1, were dysregulated in mice with carbon tetrachloride (CCl4 )-induced liver fibrosis. In turn, disruption of the circadian clock exacerbated experimental liver fibrosis. NR1D1-deficient mice were more sensitive to CCl4 -induced liver fibrosis, supporting a critical role of NR1D1 in liver fibrosis development. Validation at the tissue and cellular levels showed that NR1D1 was primarily degraded by N6-methyladenosine (m 6 A) methylation in a CCl4 -induced liver fibrosis model, and this result was also validated in rhythm-disordered mouse models. In addition, the degradation of NR1D1 further inhibited the phosphorylation of dynein-related protein 1-serine site 616 (DRP1 S616 ), resulting in weakened mitochondrial fission function and increased mitochondrial DNA (mtDNA) release in hepatic stellate cell (HSC), which in turn activated the cGMP-AMP synthase (cGAS) pathway. Activation of the cGAS pathway induced a local inflammatory microenvironment that further stimulated liver fibrosis progression. Interestingly, in the NR1D1 overexpression model, we observed that DRP1 S616 phosphorylation was restored, and cGAS pathway was also inhibited in HSCs, resulting in improved liver fibrosis. Taken together, our results suggest that targeting NR1D1 may be an effective approach to liver fibrosis prevention and management. Graphical Abstract: m 6 A methylation induced-NR1D1 ablation integrated liver clock with CCl4 -induced hepatic fibrosis through inhibiting mitochondrial fission of HSC, which activated ROS and cGAS pathway, leading to the persistent activation of HSC. ga1 Highlights: We found that the process of liver fibrosis was accompanied by obvious rhythm disturbance, and the rhythm disturbance model was more likely to cause the occurrence and development of liver fibrosis disease in mice. It was found that the persistent progression of liver fibrosis may be caused by the imbalance of cell rhythm and mitochondrial dysfunction caused by the loss of NR1D1 in hepatic stellate cells. It was found that m 6 A methylation degradation of NR1D1 might be an important mechanism for the sustained activation of hepatic stellate cells, which also provided a new intervention target for drug development.