murine-precision-cut-liver-slices-as-an-ex-vivo-model-of-liver-biology

Murine Precision-Cut Liver Slices as an Ex Vivo Model of Liver Biology

Understanding the mechanisms of liver injury, hepatic fibrosis, and cirrhosis that underlie chronic liver diseases (i.e., viral hepatitis, non-alcoholic ...

QIMR Berghofer Medical Research Institute

The University of Queensland

Madrid Institute for Advanced Studies (IMDEA) in Food, CEI UAM+CSIC

Monocyte-derived macrophages are critical in the development of atherosclerosis and can adopt a wide range of functional phenotypes depending on their surrounding milieu. High-density lipoproteins (HDLs) have many cardio-protective properties including potent anti-inflammatory effects. We investigated the effects of HDL on human macrophage phenotype and the mechanisms by which these occur.

High-density lipoprotein inhibits human M1 macrophage polarization through redistribution of caveolin-1.

Caveolin-1 (CAV1), the structural protein of caveolae in the plasma membrane, has emerged as a regulator of liver function. CAV1 modulates several molecular pathways leading to the regulation of hepatic lipid accumulation, lipid and glucose metabolism, mitochondrial biology, and hepatocyte proliferation. CAV1 thus plays a crucial role in maintaining hepatic physiology during metabolic adaptation to fasting, liver steatosis, and hepatocyte proliferation associated with liver regeneration. With failure of such processes, CAV1 has been implicated in the modulation of cholestasis, hepatitis, cirrhosis, and hepatocarcinogenesis. This review discusses the latest research in CAV1 biology and related proteins, aiming to guide future endeavors that explore their role in liver physiology and disease.

Caveolin-1 Function in Liver Physiology and Disease.

Caveolin-1 (CAV1) has emerged as a promoter of proliferation, metastasis, and chemoresistance in hepatoma cells, as well as a marker of poor prognosis in liver cancer. We discuss here current knowledge and future approaches to elucidating the molecular mechanisms underlying CAV1 action during hepatocarcinogenesis and evaluate its potential use in clinical therapies.

Filling the Gap on Caveolin-1 in Liver Carcinogenesis.

Cystic fibrosis liver disease (CFLD) in children causes progressive fibrosis leading to biliary cirrhosis, however its cause(s) and early pathogenesis are unclear. We hypothesised that a bile acid-induced Ductular Reaction (DR) drives fibrogenesis. We evaluated the DR by cytokeratin-7 immunohistochemistry in liver biopsies, staged for fibrosis, from 60 children with CFLD and demonstrated that the DR was significantly correlated with hepatic fibrosis stage and biliary taurocholate levels. To examine the mechanisms involved in DR induction liver progenitor cells (LPCs) were treated with taurocholate and key events in DR evolution were assessed: LPC proliferation, LPC biliary differentiation and hepatic stellate cell (HSC) chemotaxis. Taurocholate induced a time-dependent increase in LPC proliferation and expression of genes associated with cholangiocyte differentiation (cytokeratin-19, connexin-43, integrin-β4 and γ-glutamyltranspeptidase [GGT]), while the hepatocyte specification marker HNF4α was suppressed. Functional cholangiocyte differentiation was demonstrated via increased acetylated α-tubulin and SOX9 proteins, the number of primary cilia(+) LPCs and increased active GGT enzyme secretion. Taurocholate induced LPCs to release MCP-1, MIP1α and RANTES into conditioned medium causing HSC chemotaxis, which was inhibited by anti-MIP1α. Immunofluorescence confirmed chemokine expression localized to CK7(+) DR and LPCs in CFLD liver biopsies. This study suggests that taurocholate is involved in initiating functional LPC biliary differentiation and the development of the DR, with subsequent induction of chemokines that drive HSC recruitment in CFLD.

Taurocholate induces biliary differentiation of liver progenitor cells causing hepatic stellate cell chemotaxis in the Ductular Reaction: Role in pediatric cystic fibrosis liver disease.

Cystic fibrosis (CF)-associated liver disease (CFLD) is a hepatobiliary complication of CF. Current diagnostic modalities rely on nonspecific assessments, whereas liver biopsy is the gold standard to assess severity of fibrosis. MicroRNAs (miRNAs) regulate liver disease pathogenesis and are proposed as diagnostic biomarkers. We investigated the combined use of serum miRNAs and aspartate aminotransferase (AST) to platelet ratio (APRI) to diagnose and assess CFLD severity. This was a cross-sectional cohort study of the circulatory miRNA signature of 124 children grouped by clinical, biochemical, and imaging assessments as follows: CFLD (n = 44), CF patients with no evidence of liver disease (CFnoLD; n = 40), and healthy controls (n = 40). Serum miRNAs were analyzed using miRNA sequencing (miRNA-Seq). Selected differentially expressed serum miRNA candidates were further validated by qRT-PCR and statistical analysis performed to evaluate utility to predict CFLD and fibrosis severity validated by liver biopsy, alone or in combination with APRI. Serum miR-122-5p, miR-365a-3p, and miR-34a-5p levels were elevated in CFLD compared to CFnoLD, whereas miR-142-3p and let-7g-5p were down-regulated in CFLD compared to CFnoLD. Logistic regression analysis combining miR-365a-3p, miR-142-3p, and let-7g-5p with APRI showed 21 times greater odds of accurately predicting liver disease in CF with an area under the receiver operating characteristics curve (AUROC) = 0.91 (sensitivity = 83%, specificity = 92%; P < 0.0001). Expression levels of serum miR-18a-5p were correlated with increasing hepatic fibrosis (HF) stage in CFLD (r  = 0.56; P < 0.0001), showing good diagnostic accuracy for distinguishing severe (F3-F4) from mild/moderate fibrosis (F0-F2). A unit increase of miR-18a-5p showed a 7-fold increased odds of having severe fibrosis with an AUROC = 0.82 (sensitivity = 93%, specificity = 73%; P = 0.004), indicating its potential to predict fibrosis severity. Conclusion: We identified a distinct circulatory miRNA profile in pediatric CFLD with potential to accurately discriminate liver disease and fibrosis severity in children with CF.

MicroRNA Sequencing Identifies a Serum MicroRNA Panel, Which Combined With Aspartate Aminotransferase to Platelet Ratio Index Can Detect and Monitor Liver Disease in Pediatric Cystic Fibrosis.

Article	Year
High-density lipoprotein inhibits human M1 macrophage polarization through redistribution of caveolin-1. British journal of pharmacology\| PubMed ID: 26332942	2016
Caveolin-1 Function in Liver Physiology and Disease. Trends in molecular medicine\| PubMed ID: 27633517	2016
Filling the Gap on Caveolin-1 in Liver Carcinogenesis. Trends in cancer\| PubMed ID: 28741517	2016
Taurocholate induces biliary differentiation of liver progenitor cells causing hepatic stellate cell chemotaxis in the Ductular Reaction: Role in pediatric cystic fibrosis liver disease. The American journal of pathology\| PubMed ID: 28935574	2017
MicroRNA Sequencing Identifies a Serum MicroRNA Panel, Which Combined With Aspartate Aminotransferase to Platelet Ratio Index Can Detect and Monitor Liver Disease in Pediatric Cystic Fibrosi... Hepatology (Baltimore, Md.)\| PubMed ID: 30014495	2018
The antiproliferative and apoptotic profile of gomesin against DFTD. Cell death & disease\| PubMed ID: 30082778	2018

Manuel A. Fernandez-Rojo

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