Similar Datasets
Project description:There is a high need for predictive human ex vivo models for non-alcoholic fatty liver disease (NAFLD). About a decade ago, precision-cut liver slices (PCLSs) have been established as an ex vivo assay for humans and other organisms. In the present study, we use transcriptomics by RNASeq to profile a new human and mouse PCLSs based assay for steatosis in NAFLD. Steatosis as quantified by an increase of triglycerides after 48 h in culture, is induced by incremental supplementation of sugars (glucose and fructose), insulin, and fatty acids (palmitate, oleate). We mirrored the experimental design for human vs. mouse liver organ derived PCLSs and profiled each organ at eight different nutrient conditions after 24 h and 48 h time in culture. Thus, the provided data allows a comprehensive analysis of the donor, species, time, and nutrient factor specific regulation of gene expression in steatosis, despite the heterogeneity of the human tissue samples. Exemplified this is demonstrated by ranking homologous gene pairs by convergent or divergent expression pattern across nutrient conditions.
| S-EPMC10199090 | biostudies-literature
Project description:Progression of non-alcoholic fatty liver disease (NAFLD) to non-alcoholic steatohepatitis (NASH) is a major cause of end-stage liver diseases. There is a high need for predictive human ex vivo models to translate preclinical findings for potential new NAFLD drugs to human disease. About a decade ago, precision cut liver slices (PCLSs) have been established as an ex vivo assay for humans and other organisms. The molecular and functional baseline characteristics of PCLSs have been extensively described previously. In the present study, we provide a comprehensive RNA-Seq based molecular characterization of a new assay for NAFLD that induces steatosis in human and mouse PCLSs culture by incremental supplementation of sugars (glucose and fructose), insulin, and fatty acids (palmitate, oleate). Independent of the triglyceride levels of the human donor liver organs at baseline, the nutrient over-supplementation leads to an increase of triglyceride synthesis and cytokine release to the culture supernatant and thus recapitulates hallmark features of steatosis and inflammation in NAFLD. The mirrored design of human vs. mouse liver slices cultured at the exact same conditions allows a comprehensive assessment of donor variability and the contribution of time vs. nutrient factors to the overall observed molecular pattern and the investigation of conserved and species-specific transcriptional pathways that are involved in steatosis and inflammation of the liver.
2023-05-09 | GSE200418 | GEO
Project description:Progression of non-alcoholic fatty liver disease (NAFLD) to non-alcoholic steatohepatitis (NASH) is a major cause of end-stage liver diseases. There is a high need for predictive human ex vivo models to translate preclinical findings for potential new NAFLD drugs to human disease. About a decade ago, precision cut liver slices (PCLSs) have been established as an ex vivo assay for humans and other organisms. The molecular and functional baseline characteristics of PCLSs have been extensively described previously. In the present study, we provide a comprehensive RNA-Seq based molecular characterization of a new assay for NAFLD that induces steatosis in human and mouse PCLSs culture by incremental supplementation of sugars (glucose and fructose), insulin, and fatty acids (palmitate, oleate). Independent of the triglyceride levels of the human donor liver organs at baseline, the nutrient over-supplementation leads to an increase of triglyceride synthesis and cytokine release to the culture supernatant and thus recapitulates hallmark features of steatosis and inflammation in NAFLD. The mirrored design of human vs. mouse liver slices cultured at the exact same conditions allows a comprehensive assessment of donor variability and the contribution of time vs. nutrient factors to the overall observed molecular pattern and the investigation of conserved and species-specific transcriptional pathways that are involved in steatosis and inflammation of the liver.
2023-05-09 | GSE200408 | GEO
Project description:Transcriptomic profiling of induced steatosis in precision-cut liver slices
| PRJNA824476 | ENA
Project description:Transcriptomic profiling of induced steatosis in mouse precision-cut liver slices
| PRJNA824462 | ENA
Project description:Transcriptomic profiling of induced steatosis in human precision-cut liver slices
| PRJNA824481 | ENA
Project description:Metabolic-associated fatty liver disease (MAFLD) starts with hepatic triglyceride accumulation (steatosis) and can progress to more severe stages such as non-alcoholic steatohepatitis (NASH) and even cirrhosis. Butyrate, and butyrate-producing bacteria, have been suggested to reduce liver steatosis directly and systemically by increasing liver β-oxidation. This study aimed to examine the influence of butyrate directly on the liver in an ex vivo induced MAFLD model. To maintain essential intercellular interactions, precision-cut liver slices (PCLSs) were used. These PCLSs were prepared from male C57BL/6J mice and cultured in varying concentrations of fructose, insulin, palmitic acid and oleic acid, to mimic metabolic syndrome. Dose-dependent triglyceride accumulation was measured after 24 and 48 h of incubation with the different medium compositions. PCLSs viability, as indicated by ATP content, was not affected by medium composition or the butyrate concentration used. Under induced steatotic conditions, butyrate did not prevent triglyceride accumulation. Moreover, it lowered the expression of genes encoding for fatty acid oxidation and only increased C4 related carnitines, which indicate butyrate oxidation. Nevertheless, butyrate lowered the fibrotic response of PCLSs, as shown by reduced gene expression of fibronectin, alpha-smooth muscle actin and osteopontin, and protein levels of type I collagen. These results suggest that in the liver, butyrate alone does not increase lipid β-oxidation directly but might aid in the prevention of MAFLD progression to NASH and cirrhosis.
| S-EPMC8703944 | biostudies-literature
Project description:Hepatotoxicity is one of the major reasons for withdrawal of drugs from the market. Therefore, there is a need to screen new drugs for hepatotoxicity in humans at an earlier stage. The aim of this study was to validate human precision-cut liver slices (PCLS) as an ex vivo model to predict drug-induced cholestasis and identify the possible mechanisms of cholestasis-induced toxicity using gene expression profiles. Five hepatotoxicants, which are known to induce cholestasis (alpha-naphthyl isothiocyanate, chlorpromazine, cyclosporine, ethinyl estradiol and methyl testosterone) were used at concentrations inducing low (<30 %) and medium (30-50 %) toxicity, based on ATP content. Human PCLS were incubated with the drugs in the presence of a non-toxic concentration (60 µM) of a bile acid mixture (portal vein concentration and composition) as model for bile acid-induced cholestasis. Regulated genes include bile acid transporters and cholesterol transporters. Pathway analysis revealed that hepatic cholestasis was among the top ten regulated pathways, and signaling pathways such as farnesoid X receptor- and liver X receptor-mediated responses, which are known to play a role in cholestasis, were significantly affected by all cholestatic compounds. Other significantly affected pathways include unfolded protein response and protein ubiquitination implicating the role of endoplasmic reticulum stress. This study shows that human PCLS incubated in the presence of a physiological bile acid mixture correctly reflect the pathways affected in drug-induced cholestasis in the human liver. In the future, this human PCLS model can be used to identify cholestatic adverse drug reactions of new chemical entities.
| S-EPMC5316400 | biostudies-literature
Project description:This SuperSeries is composed of the SubSeries listed below.
2016-01-27 | GSE76163 | GEO
Project description:Drugs are often withdrawn from the market due to the manifestation of drug-induced liver injury (DILI) in patients. Drug-induced cholestasis (DIC), defined as obstruction of hepatic bile flow due to medication, is one form of DILI. Because DILI is idiosyncratic, and the resulting cholestasis complex, there is no suitable in vitro model for early DIC detection during drug development. Our goal was to develop a mouse precision-cut liver slice (mPCLS) model to study DIC and to assess cholestasis development using conventional molecular biology and analytical chemistry methods. Cholestasis was induced in mPCLS through a 48-h-incubation with three drugs known to induce cholestasis in humans, namely chlorpromazine (15, 20, and 30 µM), cyclosporin A (1, 3, and 6 µM) or glibenclamide (25, 50, and 65 µM). A bile-acid mixture (16 µM) that is physiologically representative of the human bile-acid pool was added to the incubation medium with drug, and results were compared to incubations with no added bile acids. Treatment of PCLS with cholestatic drugs increased the intracellular bile-acid concentration of deoxycholic acid and modulated bile-transporter genes. Chlorpromazine led to the most pronounced cholestasis in 48 h, observed as increased toxicity; decreased protein and gene expression of the bile salt export pump; increased gene expression of multidrug resistance-associated protein 4; and accumulation of intracellular bile acids. Moreover, chlorpromazine-induced cholestasis exhibited some transition into fibrosis, evidenced by increased gene expression of collagen 1A1 and heatshock protein 47. In conclusion, we demonstrate that mPCLS can be used to study human DIC onset and progression in a 48 h period. We thus propose this model is suited for other similar studies of human DIC.
| S-EPMC9325861 | biostudies-literature