Critical Factors in the Assessment of Cholestatic Liver Injury In Vitro

Woolbright, Benjamin L.; Jaeschke, Hartmut

doi:10.1007/978-1-4939-2074-7_28

Cited by 14 publications

(16 citation statements)

References 50 publications

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“…We assume that YAP activation has a major role in hepatocyte transdifferentiation upon induced cholestasis. To prove our hypothesis, we treated primary hepatocytes from AlbCre + Rbpj +/+ and AlbCre + Rbpj −/− mice with glycocholic acid (GCA) to mimic cholestatic liver injury in vitro [30] (Figure 6A). Since the AlfpCre mediated deletion of Rbpj had a severe phenotype, we used a different Cre driver line for this experiment.…”

Section: Resultsmentioning

confidence: 99%

YAP Activation Drives Liver Regeneration after Cholestatic Damage Induced by Rbpj Deletion

Tharehalli

Svinarenko

et al. 2018

IJMS

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Liver cholestasis is a chronic liver disease and a major health problem worldwide. Cholestasis is characterised by a decrease in bile flow due to impaired secretion by hepatocytes or by obstruction of bile flow through intra- or extrahepatic bile ducts. Thereby cholestasis can induce ductal proliferation, hepatocyte injury and liver fibrosis. Notch signalling promotes the formation and maturation of bile duct structures. Here we investigated the liver regeneration process in the context of cholestasis induced by disruption of the Notch signalling pathway. Liver-specific deletion of recombination signal binding protein for immunoglobulin kappa j region (Rbpj), which represents a key regulator of Notch signalling, induces severe cholestasis through impaired intra-hepatic bile duct (IHBD) maturation, severe necrosis and increased lethality. Deregulation of the biliary compartment and cholestasis are associated with the change of several signalling pathways including a Kyoto Encyclopedia of Genes and Genomes (KEGG) gene set representing the Hippo pathway, further yes-associated protein (YAP) activation and upregulation of SRY (sex determining region Y)-box 9 (SOX9), which is associated with transdifferentiation of hepatocytes. SOX9 upregulation in cholestatic liver injury in vitro is independent of Notch signalling. We could comprehensively address that in vivo Rbpj depletion is followed by YAP activation, which influences the transdifferentiation of hepatocytes and thereby contributing to liver regeneration.

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Section: Resultsmentioning

confidence: 99%

YAP Activation Drives Liver Regeneration after Cholestatic Damage Induced by Rbpj Deletion

Tharehalli

Svinarenko

et al. 2018

IJMS

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“…During DIC, exposure to the drug in question primarily results from the pathophysiological accumulation of bile salts. In humans, bile salt-induced toxicity is predominantly characterized by increases in serum markers of necrotic, but not apoptotic, cell death [19] . Popular examples of necrosis assays include the lactate dehydrogenase (LDH) release assay and the (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay [20 , 21] .…”

Section: Methods Validationmentioning

confidence: 99%

Drug-induced cholestasis assay in primary hepatocytes

Brantegem

Chatterjee

Bruyn³

et al. 2020

MethodsX

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Drug-induced cholestasis (DIC) is a major cause of clinical failure of drug candidates. Numerous patients worldwide are affected when exposed to marketed drugs exhibiting a DIC signature. Prospective identification of DIC during early compound development remains challenging. Here we describe the optimized in vitro procedure for early assessment and prediction of an increased DIC risk. Our method is based on three principles: Exposure of primary human hepatocyte cultures to test compounds in the absence and presence of a physiologically relevant mixture of endogenous bile salts. Rapid and quantitative assessment of the influence of concomitant bile salt exposure on hepatocyte functionality and integrity after 24 h or 48 h of incubation. Translation of the in vitro result, expressed as a DIC index (DICI) value, into an in vivo safety margin. Using our historical control data, a new (data driven) DICI cut-off value of 0.78 was established for discerning cholestatic and non-cholestatic compounds. Our DIC assay protocol was further improved by now relying on the principle of the no observable adverse effect level (NOAEL) for determining the highest test compound concentration corresponding to a DICI ≥ 0.78. Predicted safety margin values were subsequently calculated for compounds displaying hepatotoxic and/or cholestatic effects in patients, thus enabling evaluation of the performance of our DIC assay. Of note, this assay can be extended to explore the role of drug metabolites in precipitating DIC.

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“…Thus, the mouse bile acid pool is much more hydrophilic compared to the human bile acid pool. This is the main reason for the direct cytotoxicity of bile acid during cholestasis in humans but not mice [234,235]. Nevertheless, similar mechanisms of bile acid metabolism likely apply to most species.…”

Section: Bile Acids Co-regulate Post-hepatectomy Liver Regeneration Imentioning

confidence: 99%

Post-hepatectomy liver regeneration in the context of bile acid homeostasis and the gut-liver signaling axis

et al. 2018

J Clin Transl Res

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Background: Liver regeneration following partial hepatectomy (PHx) is a complicated process involving multiple organs and several types of signaling networks. The bile acid-activated metabolic pathways occupy an auxiliary yet important chapter in the entire biochemical story. PHx is characterized by rapid but transient bile acid overload in the liver, which constitutes the first wave of proliferative signaling in the remnant hepatocytes. Bile acids trigger hepatocyte proliferation through activation of several nuclear receptors. Following biliary passage into the intestines, enterocytes reabsorb the bile acids, which results in the activation of farnesoid X receptor (FXR), the consequent excretion of fibroblast growth factor (FGF)19/FGF15, and its release into the enterohepatic circulation. FGF19/FGF15 subsequently binds to its cognate receptor, fibroblast growth factor receptor 4 (FGFR4) complexed with β-klotho, on the hepatocyte membrane, which initiates the second wave of proliferative signaling. Because some bile acids are toxic, the remnant hepatocytes must resolve the potentially detrimental state of bile acid excess. Therefore, the hepatocytes orchestrate a bile acid detoxification and elimination response as a protective mechanism in concurrence with the proliferative signaling. The response in part results in the excretion of (biotransformed) bile acids into the canalicular system, causing the bile acids to end up in the intestines. Relevance for patients: Recently, FXR agonists have been shown to promote regeneration via the gut-liver axis. This type of pharmacological intervention may prove beneficial for patients with hepatobiliary tumors undergoing PHx. In light of these developments, the review provides an in-depth account of the pathways that underlie post-PHx liver regeneration in the context of bile acid homeostasis in the liver and the gut-liver signaling axis.

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Critical Factors in the Assessment of Cholestatic Liver Injury In Vitro

Cited by 14 publications

References 50 publications

YAP Activation Drives Liver Regeneration after Cholestatic Damage Induced by Rbpj Deletion

YAP Activation Drives Liver Regeneration after Cholestatic Damage Induced by Rbpj Deletion

Drug-induced cholestasis assay in primary hepatocytes

Post-hepatectomy liver regeneration in the context of bile acid homeostasis and the gut-liver signaling axis

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