Project description:This study used 10X Genomics, single-cell RNA-sequencing to examine the heterogeneity of liver non-parenchymal cell types present in the normal and CCl4-mediated fibrotic mice.
Project description:The cellular source for hepatocyte transplantation and liver regeneration in chronically hepatic diseases is a fundamental topic in liver biology. A successful therapeutic approach based on a benificial stem/progenitor cells would involve replacing damaged cells or restoring homeostasis to the areas that underlie the fibrotic response. This study aims to isolate and culture hepatocyte and non-parenchymal cells derived liver progenitor-like cells (HepLPCs and NPC-LPCs) to identify a beneficial cell sources for cell-based therapy against chronic liver injury. Two types of lineage tracing models including tdTomato mice with AAV8-TBG-Cre and AlbCreERT/R26GFP were purified for hepatocytes and non-parenchymal cells respectively, and cultured for transition to progenitor-like cells (LPCs) in TEM. The differences between them were clarified by RNA sequencing, suggesting that HepLPCs retaining basal levels of hepatic function and transcription factors (TFs), including Foxa3. Organoids and 3D hepatic spheroids culturing systems were used for further analysis of bile duct and hepatic functioning diversity. When transplanting in mice with CCL4 treatment, HepLPCs displayed the characteristics of TFs regulatory reactivation and physiological balance restoration, resulting in a significant improvement in liver fibrosis compared with NPC-LPCs, which are crucial for liver regeneration in the treatment of patients with a broad range of hepatic fibrosis.
Project description:Hepatic stellate cells are involved in the development of hepatic fibrosis. We here perform transcriptional profiling of hepatic stellate cells (HSCs) isolated from Western diet/high fructose-fed C57BL6/J mice, carbon tretrachloride (CCl4)-treated C57BL6/J mice, and of murine HSCs differentiated in vitro. Specifically, gene expression profiles are obtained from hepatic stellate cells isolated from C57BL6 mice fed a Western Diet supplemented with high fructose for 12, 16 or 24 weeks or normal chow. From hepatic stellate cells isolated from C57BL6 mice treated CCl4 for 1, 4 or 8 weeks or treated with vehicle. From hepatic stellate cells isolated from healthy C57BL6 mice and seeded on normal plastic cell culture dishes for 1, 4, 8, or 12 days. And from hepatic stellate cells isolated from healthy C57BL6 mice and seeded on normal plastic cell culture dishes for 6 days in the presence of 10uM U0126 or DMSO.
Project description:The goal of this project is to study hepatic stellate cell sub-populations in liver fibrosis and more specifically find a sub-population which responds to stiffness. Single cells were isolated from healthy versus CCl4-treated mice utilizing accudenz gradient method and were sequenced using 10X genomics technology.
Project description:Wnt/β-catenin is involved in every aspect of embryonic development and in the pathogenesis of many human diseases, and is also implicated in organ fibrosis. However, the role of β-catenin-mediated signaling on liver fibrosis remains unclear. To explore this issue, the effects of PRI-724, a selective inhibitor of the cAMP-response element-binding protein-binding protein (CBP)/β-catenin interaction, on liver fibrosis were examined using carbon tetrachloride (CCl4)- or bile duct ligation (BDL)-induced mouse liver fibrosis models. Following repetitive CCl4 administrations, the nuclear translocation of β-catenin was observed only in the non-parenchymal cells in the liver. PRI-724 treatment reduced the fibrosis induced by CCl4 or BDL, accompanied by the suppression of S100A4 expression, a CBP/β-catenin transcript. C-82, an active form of PRI-724, inhibited the activation of isolated primary mouse quiescent hepatic stellate cells (HSCs) and promoted cell death in culture-activated HSCs. During the fibrosis resolution period, an increase in F4/80+ CD11b+ and Ly6Clow CD11b+ macrophages was induced by CCl4 and was sustained for two weeks thereafter, even after having stopped CCl4 treatment. PRI-724 accelerated the resolution of CCl4-induced liver fibrosis, and this was accompanied by increased matrix metalloproteinase (MMP)-9, MMP-2, and MMP-8 expression in intrahepatic leukocytes. These results suggest that the inhibition of CBP/β-catenin suppresses liver fibrosis through the inhibition of HSCs activation, the induction of activated HSC death, and the production of MMPs from macrophages. Thus, targeting the CBP/β-catenin interaction may become a new therapeutic strategy in treating liver fibrosis. We used microarrays to detail the global change of gene expression by PRI-724 (C-82)-treatment in culture-activated murine hepatic stellate cells.
Project description:Norm mouse hepatocyte-derived extracellular vesicles were used to treat CCl4-injured mice and the hepatic transcript changes were measured by RNA-seq.
Project description:Gene expression was measured by scRNAseq in isolated liver cells from mice treated with CCl4 and in non parenchymal Col1a1+ cells from a 3 month old Mdr2KO mouse.
Project description:This study aimed to investigate the protective mechanisms of helenalin on hepatic fibrosis,Rats were intragastrically administrated with 50% CCl4 for 9 weeks to induce liver fibrosis, followed by various medicines for 6 weeks. The transcriptomic analysis was performed in liver tissues by RNA-seq.