Sorafenib reduces steatosis‐induced fibrogenesis in a human 3D co‐culture model of non‐alcoholic fatty liver disease

• Published in: Environmental toxicology 2021-02, Vol.36 (2), p.168-176
• Main Authors: Romualdo, Guilherme Ribeiro, Da Silva, Tereza Cristina, Landi, Marina Frota, Morais, Juliana Ávila, Barbisan, Luis Fernando, Vinken, Mathieu, Oliveira, Cláudia Pinto, Cogliati, Bruno
• Format: Article
• Language: English
• Subjects: Animal models; Animals; antifibrotic therapy; Balloon treatment; Cell activation; Cell culture; Cell Line, Tumor; Cell Survival - drug effects; Cells; Chemical compounds; Cirrhosis; Coculture Techniques; Collagen (type I); fatty hepatocytes; Fatty liver; Fibrosis; Genes; Health services; hepatic stellate cells; Hepatic Stellate Cells - drug effects; Hepatic Stellate Cells - pathology; Hepatocytes; Hepatocytes - drug effects; Hepatocytes - pathology; Hepatoma; Human diseases; Humans; Inhibitor drugs; Lipid metabolism; Lipid peroxidation; Lipids; Liver; Liver - drug effects; Liver - metabolism; Liver - pathology; Liver cancer; Liver cirrhosis; Liver Cirrhosis - metabolism; Liver Cirrhosis - pathology; Liver Cirrhosis - prevention & control; Liver diseases; Metabolism; Microenvironments; Non-alcoholic Fatty Liver Disease - drug therapy; Non-alcoholic Fatty Liver Disease - metabolism; Non-alcoholic Fatty Liver Disease - pathology; non‐alcoholic steatohepatitis; Oleic acid; Oxidation; Pharmacology; Platelet-derived growth factor; Sorafenib - pharmacology; Spheroids; Steatosis; Three dimensional models; Tissue inhibitor of metalloproteinase 1; Tissue Inhibitor of Metalloproteinase-1 - metabolism; Transforming Growth Factor beta1 - metabolism; Transforming growth factor-b1; tridimensional cell culture; Tumor necrosis factor; Two dimensional models
• ISSN: 1520-4081; 1522-7278
• DOI: 10.1002/tox.23021

Non‐alcoholic fatty liver disease (NAFLD) affects around 25% of the worldwide population. Non‐alcoholic steatohepatitis (NASH), the more progressive variant of NAFLD, is characterized by steatosis, cellular ballooning, lobular inflammation, and may culminate on hepatic stellate cell (HSC) activation, thus increasing the risk for fibrosis, cirrhosis, and HCC development. Conversely, the antifibrotic effects of sorafenib, an FDA‐approved drug for HCC treatment, have been demonstrated in 2D cell cultures and animal models, but its mechanisms in a NAFLD‐related microenvironment in vitro requires further investigation. Thus, a human 3D co‐culture model of fatty hepatocytes and HSC was established by culturing hepatoma C3A cells, pre‐treated with 1.32 mM oleic acid, with HSC LX‐2 cells. The fatty C3A/LX‐2 spheroids showed morphological and molecular hallmarks of altered lipid metabolism and steatosis‐induced fibrogenesis, similarly to the human disease. Sorafenib (15 μM) for 72 hours reduced fatty spheroid viability, and upregulated the expression of lipid oxidation‐ and hydrolysis‐related genes, CPT1 and LIPC, respectively. Sorafenib also inhibited steatosis‐induced fibrogenesis by downregulating COL1A1, TGFB1, PDGF, and TIMP1 and by decreasing protein levels of IL‐6, TGF‐β1, and TNF‐α in fatty spheroids. The demonstration of the antifibrotic properties of sorafenib on steatosis‐induced fibrogenesis in a 3D in vitro model of NAFLD supports its clinical use as a therapeutic agent for the treatment of NAFLD/NASH patients.