Neutrophils interact with cholangiocytes to cause cholestatic changes in alcoholic hepatitis

Takeuchi, Masahiro; Vidigal, P.T.; Guerra, Mateus T.; Hundt, Melanie; Robert, Marie E.; Olave-Martinez, Maria; Aoki, Satoshi; Khamphaya, Tanaporn; Kersten, Remco; Kruglov, Emma A.; Rodríguez, Randolph de la Rosa; Nathanson, Michael H.; Weerachayaphorn, Jittima

doi:10.1136/gutjnl-2020-322540

Cited by 30 publications

(51 citation statements)

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“…Cholangiocytes play a previously unrecognized role in the development of cholestasis in alcoholic hepatitis. 28 We tested ethanol-associated biliary injury in vitro, using hICOs and ALD-ICOs, which potentially are associated with alcoholic liver disease etiology ( Figure 6 A ). The nonoxidative ethanol metabolite palmitoleic acid (POA) has been implicated as a critical mediator in ethanol-evoked epithelial cell injury.…”

Section: Resultsmentioning

confidence: 99%

Recapitulating Cholangiopathy-Associated Necroptotic Cell Death In Vitro Using Human Cholangiocyte Organoids

Shi

Verstegen

Roest

et al. 2022

Cellular and Molecular Gastroenterology and Hepatology

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Background & Aims Liver and bile duct diseases often are associated with extensive cell death of cholangiocytes. Necroptosis represents a common mode of programmed cell death in cholangiopathy, however, detailed mechanistic knowledge is limited owing to the lack of appropriate in vitro models. To address this void, we investigated whether human intrahepatic cholangiocyte organoids (ICOs) can recapitulate cholangiopathy-associated necroptosis and whether this model can be used for drug screening. Methods We evaluated the clinical relevance of necroptosis in end-stage liver diseases and liver transplantation by immunohistochemistry. Cholangiopathy-associated programmed cell death was evoked in ICOs derived from healthy donors or patients with primary sclerosing cholangitis or alcoholic liver diseases by the various stimuli. Results The expression of key necroptosis mediators, receptor-interacting protein 3 and phosphorylated mixed lineage kinase domain-like, in cholangiocytes during end-stage liver diseases was confirmed. The phosphorylated mixed lineage kinase domain-like expression was etiology-dependent. Gene expression analysis confirmed that primary cholangiocytes are more prone to necroptosis compared with primary hepatocytes. Both apoptosis and necroptosis could be specifically evoked using tumor necrosis factor α and second mitochondrial-derived activator of caspases mimetic, with or without caspase inhibition in healthy and patient-derived ICOs. Necroptosis also was induced by ethanol metabolites or human bile in ICOs from donors and patients. The organoid cultures further uncovered interdonor variable and species-specific drug responses. Dabrafenib was identified as a potent necroptosis inhibitor and showed a protective effect against ethanol metabolite toxicity. Conclusions Human ICOs recapitulate cholangiopathy-associated necroptosis and represent a useful in vitro platform for the study of biliary cytotoxicity and preclinical drug evaluation.

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Section: Resultsmentioning

confidence: 99%

Recapitulating Cholangiopathy-Associated Necroptotic Cell Death In Vitro Using Human Cholangiocyte Organoids

Shi

Verstegen

Roest

et al. 2022

Cellular and Molecular Gastroenterology and Hepatology

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“…AH is a potentially lethal complication of alcoholic liver disease, which has been attributed to hepatocellular damage in the past 5 . Recently, binding of neutrophils to ITGB1 expressed on the cell surface of cholangiocytes was shown to contribute to epithelial cell damage and the development of cholestasis in AH 7 . Other studies showed that cholestatic liver injury can be involved in the pathogenesis of AH, moreover, impaired secretion by cholangiocytes, or cholestasis, results in a worse outcome 6 .…”

Section: Discussionmentioning

confidence: 99%

“…Recent studies showed that cholestatic liver injury can be involved in the pathogenesis of AH, moreover, impaired secretion by cholangiocytes, or cholestasis, results in a worse outcome 6 . Takeuchi et al highlighted that integrin beta-1 (ITGB1)-mediated binding of neutrophils to cholangiocytes in AH results in the development of cholestasis 7 . It is well-established that cholangiocyte secretion largely depends on the proper function of the apically expressed CFTR 8 .…”

Section: Introductionmentioning

confidence: 99%

Impaired Regulation of PMCA Activity by Defective CFTR Expression Promotes Epithelial Cell Damage in Alcoholic Pancreatitis and Hepatitis

Madácsy

Varga

Papp

et al. 2022

Preprint

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Background and aims. Alcoholic pancreatitis and hepatitis are frequent, potentially lethal diseases with limited treatment options. Our previous study reported that the expression of CFTR Cl- channel is impaired by ethanol in pancreatic ductal cells leading to more severe alcohol-induced pancreatitis. In addition to determining epithelial ion secretion, CFTR has multiple interactions with other proteins, which may influence intracellular Ca2+ signaling. Thus, we aimed to investigate the impact of ethanol-mediated CFTR damage on intracellular Ca2+ homeostasis in pancreatic ductal epithelial cells and cholangiocytes.Methods. Human and mouse pancreas and liver samples and ex vivo organoids were used to study ion secretion, intracellular signaling and protein expression and interaction. The effect of PMCA4 inhibition was analysed in a mouse model of alcohol-induced pancreatitis.Results. The decreased CFTR expression impaired PMCA function and resulted in sustained intracellular Ca2+ elevation in ethanol-treated and mouse and human pancreatic organoids. Liver samples derived from alcoholic hepatitis patients and ethanol-treated mouse liver organoids showed decreased CFTR expression and function, and impaired PMCA4 activity. PMCA4 co-localizes and physically interacts with CFTR on the apical membrane of polarized epithelial cells, where CFTR-dependent calmodulin recruitment determines PMCA4 activity. The sustained intracellular Ca2+ elevation in the absence of CFTR inhibited mitochondrial function and was accompanied with increased apoptosis in pancreatic epithelial cells and PMCA4 inhibition increased the severity of alcohol-induced AP in mice.Conclusion. Our results suggest that improving Ca2+ extrusion in epithelial cells may be a potential novel therapeutic approach to protect the exocrine pancreatic function in alcoholic pancreatitis and prevent the development of cholestasis in alcoholic hepatitis.

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“…Cholestasis is the intrahepatic accumulation of toxic bile acids that occurs in several liver diseases, which were caused by various factors, such as pregnancy, drugs, alcohol, etc (Chatterjee and Annaert, 2018;Ovadia et al, 2021;Takeuchi et al, 2021). The occurrence of cholestatic liver diseases is very rare, with an annual incidence of between 0.3/100000 and 5.8/100000 (Wagner and Fickert, 2020).…”

Section: Discussionmentioning

confidence: 99%

Mechanism of Paeoniflorin on ANIT-Induced Cholestatic Liver Injury Using Integrated Metabolomics and Network Pharmacology

et al. 2021

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Background: Paeoniflorin (PF), the major active compound isolated from the roots of Paeonia lactiflora Pall., has been used in the treatment of severe hepatic diseases for several decades and displays bright prospects in liver protective effect. However, its biological mechanism that regulates bile acid metabolism and cholestatic liver injury has not been fully elucidated. Our study aims to investigate the mechanism by which PF in the treatment of cholestatic liver injury using a comprehensive approach combining metabolomics and network pharmacological analysis.Methods: The hepatoprotective effect of PF against cholestasis liver injury, induced by α-naphthylisothiocyanate (ANIT), was evaluated in rats. The serum biochemical indices including ALT, AST, TBA, TBIL, ALP, ALB, and the pathological characteristics of the liver were analyzed. Moreover, UHPLC-Q-TOF was performed to explore the feces of rats with ANIT-induced cholestatic liver injury treated with PF and the potential biomarkers were screened by metabolomics. The targets for the regulation of potential biomarkers by PF were screened by network pharmacology, and then the relevant key targets were verified by immunohistochemical and western blotting methods.Results: PF significantly improved serum indexes and alleviated liver histological damage. Metabolomics analyses showed that the therapeutic effect of PF is mainly associated with the regulation of 13 metabolites involved in 16 metabolic pathways. The “PF-targets-metabolites” interaction network was constructed, and then five key targets including CDC25B, CYP2C9, MAOB, mTOR, and ABCB1 that regulated the potential biomarkers were obtained. The above five targets were further verified by immunohistochemistry and western blotting, and the results showed that PF significantly improved the expression of key proteins regulating these biomarkers.Conclusion: Our study provides direct evidence for the modulatory properties of PF treatment on ANIT-induced cholestatic liver injury using metabolomics and network pharmacology analyses. PF exhibits favorable pharmacological effect by regulating related signal pathways and key targets for biomarkers. Therefore, these findings may help better understand the complex mechanisms and provide a new and effective approach to the treatment of cholestatic liver injury.

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Neutrophils interact with cholangiocytes to cause cholestatic changes in alcoholic hepatitis

Cited by 30 publications

References 50 publications

Recapitulating Cholangiopathy-Associated Necroptotic Cell Death In Vitro Using Human Cholangiocyte Organoids

Recapitulating Cholangiopathy-Associated Necroptotic Cell Death In Vitro Using Human Cholangiocyte Organoids

Impaired Regulation of PMCA Activity by Defective CFTR Expression Promotes Epithelial Cell Damage in Alcoholic Pancreatitis and Hepatitis

Mechanism of Paeoniflorin on ANIT-Induced Cholestatic Liver Injury Using Integrated Metabolomics and Network Pharmacology

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