Journal of Hepatology

Editorial Board

2010-10-01

Double announcement:Consensus Conference/Basic school

2010-10-01

Monothematic Conference, Athens, Greece

2010-10-01

Fellowship programme

2010-10-01

International Liver Congress, Berlin, Germany

2010-10-01

Focus

Daniel Shouval — 2010-07-02

Fibrosis of the liver is a complex and progressive process initiated and maintained by a large number of stimuli. Significant progress has been made in the past two decades in elucidating the mechanism(s) of fibrogenesis and exploring old and new therapeutic modalities to slow down and possibly reverse ongoing scarring of the liver . Various strategies to prevent, minimize, and even revert hepatic fibrosis have been and still are being evaluated in bile duct ligated (BDL) or CCL4 treated rat model systems as well as in humans, including removal of the injurious stimuli, suppressing inflammation, down-regulating stellate cell activation and increasing apoptosis of Ito cells. Hepatic stellate cells (HSC) have emerged as an attractive target for controlling fibrosis. Proliferation of HSC which acquire a myofibroblast phenotype and synthesize increasing amounts of extracellular matrix is considered a cornerstone in generation of hepatic fibrosis. Already in 1994, Mallat and co-workers have shown in vitro that simvastatin inhibits hydroxyl-methylglutaryl-coenzyme A reductase activity and myofibroblast-like Ito cells, isolated from normal human liver, independent of its lipid lowering effects . At that time it was still unknown if this effect also exists in vivo. Since then, several reports appeared describing the inhibitory effects of statins on hepatic inflammatory activity in vitro , portal hypertension , and indirectly on hepatic fibrosis . Statins have also been reported to have an anti-fibrotic activity in non-hepatic disorders including pulmonary and renal fibrosis .

Prognosis indicator in acute liver failure: Is there a place for cell death markers?

Didier Samuel, Philippe Ichai — 2010-06-29

Acute liver failure (ALF) is an intriguing disease. The mechanisms by which liver cells are destroyed in just a matter of a few days, as well as the mechanisms by which the liver can regenerate itself in a few hours or days, remain partly unknown. It is unclear why some patients with the same apparent degree of severity and the same aetiology of liver failure have different outcomes. Indeed after the occurrence of encephalopathy, ALF patients can die without liver transplantation while others will recover either in a few hours or in a few days. Part of the complexity is due to the fact that ALF can occur through distinct pathways according to aetiologies; there is probably more than one mechanism responsible for ALF. Liver transplantation was a breakthrough in the treatment of ALF and has saved the lives of several thousands of patients who were at high risk of mortality . However, the advent of emergency liver transplantation as a potential treatment for ALF has highlighted the need for prognosis indicators. Indeed, there is a need to establish the prognosis of patients with ALF who are at risk of mortality without liver transplantation, and also for those who will survive spontaneously or with medical care only. An additional difficulty is to have reliable criteria of prognosis early in the course of ALF in order to have sufficient time to obtain a liver graft. Assessment of the prognosis of ALF is essential when making decisions on the requirement for and timing of liver transplantation. If liver transplantation is performed too early, it may be performed when it is not necessary, and if it is performed too late, there is an increased risk of the condition worsening (neurological, infectious) with a poor outcome. The prognosis of ALF depends on many factors (gender, age, cause of liver failure, hepatic, clinical and biological status on admission and at the peak of deterioration, degree of hepatic encephalopathy, prothrombin time, factor V, INR, renal function, bilirubin level, arterial pH, lactate level, phosphoremia), and many new markers could be incorporated into prognostic models.

Exploiting liver immunity for the prevention of hepatic metastases

Licia Rivoltini, Vincenzo Mazzaferro — 2010-06-21

Over the last decade immunotherapy has progressively gained a significant clinical interest for cancer treatment. The actual benefit of active immunotherapy, namely cancer vaccines, has been repeatedly claimed as effective in several settings and prompted prospective investigations (phase III clinical trials) currently ongoing in patients with prostate carcinoma, melanoma, and lung cancer, while the first vaccine for the treatment of metastatic prostate carcinoma has been recently approved by the FDA .

A phase I clinical trial of dendritic cell immunotherapy in HCV-infected individuals

2010-06-29

Background & Aims: HCV patients who fail conventional interferon-based therapy have limited treatment options. Dendritic cells are central to the priming and development of antigen-specific CD4+ and CD8+ T cell immunity, necessary to elicit effective viral clearance. The aim of the study was to investigate the safety and efficacy of vaccination with autologous dendritic cells loaded with HCV-specific cytotoxic T cell epitopes.Methods: We examined the potential of autologous monocyte-derived dendritic cells (MoDC), presenting HCV-specific HLA A2.1-restricted cytotoxic T cell epitopes, to influence the course of infection in six patients who failed conventional therapy. Dendritic cells were loaded and activated ex vivo with lipopeptides. In this phase 1 dose escalation study, all patients received a standard dose of cells by the intradermal route while sequential patients received an increased dose by the intravenous route.Results: No patient showed a severe adverse reaction although all experienced transient minor side effects. HCV-specific CD8+ T cell responses were enumerated in PBMC by ELIspot for interferon-γ. Patients generated de novo responses, not only to peptides presented by the cellular vaccine but also to additional viral epitopes not represented in the lipopeptides, suggestive of epitope spreading. Despite this, no increases in ALT levels were observed. However, the responses were not sustained and failed to influence the viral load, the anti-HCV core antibody response and the level of circulating cytokines.Conclusions: Immunotherapy using autologous MoDC pulsed with lipopeptides was safe, but was unable to generate sustained responses or alter the outcome of the infection. Alternative dosing regimens or vaccination routes may need to be considered to achieve therapeutic benefit.

The novel immunoregulatory molecule FGL2: A potential biomarker for severity of chronic hepatitis C virus infection

2010-06-24

Background & Aims: This report describes the use of a novel sensitive and specific ELISA for the measurement of human fibrinogen-like protein 2 (FGL2/fibroleukin), a novel effector of natural regulatory T (Treg) cells, to predict the course of chronic hepatitis C viral infection (HCV).Methods: Plasma levels of FGL2 were measured in HCV patients and compared to healthy controls and to patients with alcoholic liver disease.Results: FGL2 levels were significantly higher in HCV patients (84.3±89.1ng/ml, n=80) compared to healthy controls (36.4±21.9ng/ml, n=30, p<0.001), to a subset of patients who cleared HCV following anti-viral treatment (16.6±19.7ng/ml, n=32, p<0.001), and to patients with inactive alcoholic liver disease (18.8±17.4ng/ml, n=24, p<0.001). Among HCV patients, plasma levels of FGL2 correlated significantly with the stage of fibrosis (p=0.001) and were significantly higher in patients with cirrhosis (164.1+121.8ng/ml, n=60) compared to non-cirrhotics (57.7±52.8ng/ml, n=20, p=0.001). Genotype 1 patients had significantly higher levels of FGL2 (98.1±100.3ng/ml, n=60) compared to patients with genotype 2/3 (41.5±38.6ng/ml, n=20, p=0.0008). Patients with genotype 2/3 had FGL2 levels similar to healthy controls (41.5±38.6 vs. 36.41±21.9ng/ml, p=ns). Infiltrating lymphocytes in liver biopsies of HCV patients were positive for either FGL2 or FoxP3 (a marker of Treg cells) or expressed both markers.Conclusions: This report documents the development of a sensitive ELISA for measurement of plasma levels of FGL2 an effector Treg cells, which correlates with the severity of HCV infection.

Low virological response and high relapse rates in hepatitis C genotype 1 patients with advanced fibrosis despite adequate therapeutic dosing

2010-06-23

Background & Aims: The impact of fibrosis stage on chronic hepatitis C virus (HCV) treatment response was explored in CHARIOT, a study of high dose peginterferon alfa-2a (PEG-IFNα-2a) induction therapy in treatment naïve genotype 1 infection.Methods: Eight hundred and ninety-six patients were randomised 1:1 to 360μg (n=448) or 180μg (n=448) PEG-IFNα-2a weekly with RBV 1000–1200mg/day for 12weeks followed by 36weeks of 180μg PEG-IFNα-2a weekly plus RBV 1000–1200mg/day. Virological responses were assessed at week 4, 8, 12, 24, 48 (end of therapy), and 24weeks following therapy (sustained virological response, SVR). As previously reported, there was no significant difference in SVR in the induction (53%) and standard (50%) arms, therefore the pooled study population was used for analysis of SVR and relapse.Results: A marked step-wise decline in SVR was evident by fibrosis stage: F0 (70%); F1 (60%); F2 (51%); F3 (31%); F4 (10%) (p<0.0001). Early virological responses were lower among F3/4 patients, including rapid virological response (RVR) (21% vs. 34% for F3/4 and F0–2, respectively) (p=0.0072), and the RVR positive predictive value was also lower (63% vs. 80%). Virological relapse rates were similar in early disease stages (F0, 16%; F1, 23%; F2, 26%), but increased markedly in advanced fibrosis (F3, 50%; F4, 80%) (p<0.0001). Cumulative PEG-IFNα-2a and ribavirin doses were similar among patients with F3/4 and F0–2 within treatment arms through week 4, 8, 12, and week 24.Conclusions: Low virological response in hepatitis C genotype 1 patients with advanced fibrosis is not explained by inadequate cumulative PEG-IFN and ribavirin doses.

Measurement of hepatic vein pressure gradient in children with chronic liver diseases

2010-06-23

Background & Aims: The aim of this study is to present our preliminary experience with Hepatic Vein Pressure Gradient (HVPG) measurements in pediatric patients with chronic liver disease.Methods: Institutional review board approval was obtained. HVPG was measured in 20 pediatric patients, mean age 82±54months, with chronic liver disease, without extrahepatic portal vein obstruction. In nine patients the end-stage liver disease was secondary to biliary atresia; in the remaining 11, to various causes. Eleven patients had esophageal varices at endoscopy, 14 had perigastric and periesophageal collaterals at imaging scan, three had ascites, 12 had low platelet count, and all had splenomegaly.Results: Hepatic vein catheterization was technically possible in all patients without complications. HVPG values were elevated in all but three patients, ranging between 2 and 33mmHg (mean 11.3±7.2mmHg), thus indicating a sinusoidal component in portal hypertension. A salient finding was the presence of hepatic venovenous shunts in 7 out of 9 patients with biliary atresia; however, the HVPG could still be measured distal to the shunts, but in three patients (with an HVPG of 8mmHg) it was determined in an area with a small venovenous communication still visible, therefore underestimating the actual portal pressure gradient. No venovenous shunts were detected in the non-biliary atresia patients.Conclusions: HVPG is a feasible procedure in pediatric patients. Patients with biliary atresia very frequently have communicating vessels between hepatic veins. This hitherto unacknowledged finding can lead to the underestimation of portal pressure by HVPG measurement.

Prediction of oesophageal varices in hepatic cirrhosis by simple serum non-invasive markers: Results of a multicenter, large-scale study

2010-06-16

Background & Aims: Preliminary data suggest that non-invasive methods could be useful to assess presence of oesophageal varices (OV) in cirrhotic patients. We aimed to further investigate simple serum non-invasive markers for diagnosing and grading OV.Methods: A retrospective set of 510 cirrhotics and a prospective set of 110 cirrhotics were enrolled consecutively in five centers. Platelets, AST-to-ALT ratio, AST-to-platelet-ratio index, Forns’ index, Lok index, Fib-4, and Fibroindex were measured within 2months from upper endoscopy, taken as a gold standard. Performance was expressed as sensitivity, specificity, positive, and negative predictive values (PPV, NPV), accuracy, and area under the curve (AUC).Results: A combination of Lok index (cutoff=1.5) and Forns’ index (cutoff=8.8) had 0.80 AUC (0.76–0.84, 95% CI), and high NPV (>90%) to exclude clinically relevant OV, defined as large OV or small OV with red signs or in Child–Pugh C cirrhosis. By applying this combination, upper endoscopy would have been avoided in 1/3 of our cirrhotics. Large OV could be excluded with 96% NPV by Lok index (cutoff=1.5). A combination of Lok index (cutoff=0.9) and Forns’ index (cutoff=8.5) predicted presence of any grade OV with good performance: 0.82 AUC (0.76–0.88, 95% CI), 88% PPV.Conclusions: Serum non-invasive markers may be useful as a first line tool to identify cirrhotic patients in which the risk of clinically relevant OV is trivial, and to reduce the number of upper endoscopies. However, we are still far from the possibility of replacing upper endoscopy by simple serum non-invasive markers in the vast majority of patients.

Cytokeratin 18-based modification of the MELD score improves prediction of spontaneous survival after acute liver injury

2010-07-01

Background & Aims: Predicting the probability of patients with acute liver failure (ALF) to recover spontaneously is of major clinical importance. As apoptotic and necrotic cell death are crucial in the pathogenesis of ALF, we determined whether selected cell-death markers predict outcome of patients with ALF and/or discriminate between etiologies.Methods: In a prospective study (11/2006–06/2009), 68 ALF patients were recruited consecutively. Data were collected over four weeks or until discharge, death or LTx, including CK18/M65 and M30 ELISA and glutathione S-transferase, subtype α. Data at date of admission and at the date of peak levels of M65 were individually analyzed and correlated with the patients’ prognosis and etiology.Results: The predictive sensitivity of total serum M65 for lethal outcome was comparable to the Model for End-Stage Liver Disease (MELD) score at time of admission and at its peak value. In contrast, serum bilirubin levels had no prognostic value, neither at admission nor at later time points. In order to accurately predict the clinical prognosis of ALF patients, we tested a modified MELD score where CK18 M65 substituted bilirubin. This CK18/M65-based MELD score significantly better predicted the prognosis of ALF patients compared with the current MELD score or KCC. A combination of tested parameters contributed to improved discrimination of ALF etiologies by applying cell death and established laboratory parameters.Conclusions: The CK18 M65-based MELD score has superior sensitivity and specifically predicts survival of ALF patients. Further prospective clinical studies could validate its potential role to predict requirement of LTx in ALF patients.

Activation of human liver sinusoidal endothelial cell by human platelets induces hepatocyte proliferation

2010-06-23

Background & Aims: We previously reported that platelets promote hepatocyte proliferation. In this study, we focused on the role of platelets in liver sinusoidal endothelial cells (LSECs) in addition to their role in hepatocyte in liver regeneration.Methods: Immortalized human LSECs (TMNK-1) were used. The LSECs were co-cultured with human platelets, and the proliferation of LSECs and the excretion of growth factors and interleukin-6 (IL-6) were subsequently measured. The main factor from platelets which induced the excretion of IL-6 from LSECs was determined using inhibitors of each component contained in the platelets. The need for direct contact between platelets and LSECs was investigated using cell culture inserts. The proliferation of human primary hepatocytes was measured after the addition of the supernatant of LSECs cultured with or without platelets.Results: The number of LSECs cocultured with platelets significantly increased. Excretion of IL-6 and vascular endothelial growth factor (VEGF) increased in LSECs with platelets. JTE-013, a specific antagonist for sphingosine 1-phosphate (S1P) 2 receptors, inhibited the excretion of IL-6 from LSECs after the addition of platelets. When the platelets and LSECs were separated by the cell culture insert, the excretion of IL-6 from LSECs was decreased. DNA synthesis was significantly increased in human primary hepatocytes cultured with the supernatant of LSECs with platelets.Conclusions: Platelets promote LSEC proliferation and induce IL-6 and VEGF production. Direct contact between the platelets and LSECs and S1P, that are contained in platelets, were involved in the excretion of IL-6 from LSECs. IL-6 from LSECs induced proliferation of parenchymal hepatocytes.

Early growth response-1 attenuates liver injury and promotes hepatoprotection after carbon tetrachloride exposure in mice

2010-06-18

Background & Aims: Inflammatory gene expression plays a pathological role in acute and chronic hepatic inflammation, yet, inflammation also promotes liver repair by inducing protective mechanisms to limit collateral tissue damage by priming hepatocytes for proliferation. Early growth response (Egr)-1, a transcription factor that regulates inflammatory gene expression, plays a pathological role in many animal models of acute and chronic inflammatory disease. Here, we tested the hypothesis that Egr-1 is beneficial after toxic liver injury.Methods: Acute liver injury was induced in wild-type and egr-1−/− mice by a single injection of carbon tetrachloride (CCl4). Liver injury, inflammatory, and hepatoprotective gene expression and signaling events were measured 18, 48, and 72h after CCl4 administration.Results: Peak liver injury was greater in egr-1−/− mice compared to wild-type mice. Enhanced injury in egr-1−/− mice was associated with reduced tumor necrosis factor (TNF)α mRNA and protein expression, reduced Akt phosphorylation and nuclear localization of NFκB-p65 in nuclei of cells in the hepatic sinusoid. Expression of inducible nitric oxide synthase and cyclooxygenase-2, TNFα-regulated genes that have hepatoprotective function, was attenuated in egr-1−/− mice compared to wild-type mice. Although plasma interleukin (IL)-6 protein and hepatic accumulation of IL-6, glycoprotein 130, and IL-6 receptor α mRNA in wild-type and egr-1−/− mice were equivalent, signal transducer and activator of transcription 3 phosphorylation was attenuated in egr-1−/− mice and associated with reduced oncostatin M expression.Conclusions: In contrast to its role in inflammation-mediated tissue injury in other models, Egr-1 expression promotes protection in the liver after CCl4 exposure.

Pancreatic Duodenal Homeobox-1 de novo expression drives cholangiocyte neuroendocrine-like transdifferentiation

2010-06-25

Background & Aims: Reactive cholangiocytes acquire a neuroendocrine-like phenotype, with synthesis and local release of neuropeptides and hormones. The mechanism that drives such phenotypical changes is still undefined. Pancreatic Duodenal Homeobox-1 (PDX-1) is a transcription factor required for pancreatic development, that sustains pancreatic beta-cell response to injury and insulin synthesis. PDX-1 induces neuroendocrine-like transition of pancreatic ductal cells. Cholangiocyte response to injury is modulated by Glucagon-Like Peptide-1 Receptor (GLP-1R), which, in the pancreas, activates PDX-1. We wanted to verify whether PDX-1 plays any role in cholangiocyte neuroendocrine-like transdifferentiation in response to injury.Methods: PDX-1 expression was assessed in cholangiocytes from normal and one week bile duct ligated (BDL) rats. Changes in PDX-1 expression and activation upon GLP-1R activation were then assayed. The effects of the lack of PDX-1 in cholangiocytes were studied in vitro by siRNA and in vivo by the employment of PDX-1-deficient (+/−) mice.Results: BDL but not normal cholangiocytes express PDX-1. GLP-1R activation elicits, in a PI3K-dependent fashion, PDX-1 expression, together with its nuclear translocation. In vitro, GLP-1R-induced increases in VEGF and IGF-1 mRNA expression were blunted in cells with PDX-1 siRNA. In vivo, the VEGF and IGF-1 mRNA expression in the liver after one week BDL was markedly reduced in PDX-1-deficient mice, together with reduced bile duct mass.Conclusions: In response to injury, reactive cholangiocytes de novo express PDX-1, the activation of which allows cholangiocytes to synthesize IGF-1 and VEGF. These findings suggest that PDX-1 drives the acquisition of the neuroendocrine-like phenotype by cholangiocytes in response to cholestatic injury.

Genetic variations of hepatitis B virus and serum aflatoxin-lysine adduct on high risk of hepatocellular carcinoma in Southern Guangxi, China

Li Xu, Guoqing Qian, Lili Tang, Jianjia Su, Jia-Sheng Wang — 2010-06-29

Background & Aims: Southern Guangxi area is one of the endemic areas for hepatocellular carcinoma (HCC) in China. This study evaluates the roles of genetic variations of hepatitis B virus (HBV) and aflatoxin B1 (AFB1) exposure in the formation of HCC in this high-risk area.Methods: The study recruited 60 HCC patients and 120 age-, gender-, and residency-matched controls. HBV genotype and basic core promoter (BCP) mutations were determined by nested-PCR/direct sequencing. Serum AFB1-lysine adduct was measured by high performance liquid chromatography-fluorescence detection.Results: HBV genotype C was predominant in 75.0% of cases and 84.2% of controls. The 1762T/1764A double mutations, 1753V mutations, and 1752V mutations were associated with HCC risk evidenced by the adjusted odds ratio (OR) [95% confidence interval (95% CI)] of 3.89 (1.40–10.77), 2.87 (1.49–5.49), and 5.96 (1.75–20.25), respectively. The adjusted OR (95% CI) was 6.94 (1.68–27.78) for subjects with 1762T/1764A double mutations and high AFB1-lysine adduct level; 2.01 (0.24–14.29), for those with only 1762T/1764A double mutations; and 4.26 (1.16–15.38) for those with only high AFB1-lysine adduct level, respectively. The adjusted OR was 5.13 (1.79–14.71) for subjects with 1753V mutations and high AFB1-lysine adduct level; 1.20 (0.47–3.08) for those with only 1753V mutations, and 2.28 (1.01–5.31) for those with high AFB1-lysine adduct level, respectively.Conclusions: These data confirmed the association of BCP mutations with HCC risk and the additive effects of 1762T/1764A double mutations and 1753V mutations with dietary AFB1 exposure in this high-risk area for HCC.

Experimentally induced liver metastases from colorectal cancer can be prevented by mononuclear phagocyte-mediated monoclonal antibody therapy

2010-06-25

Background & Aims: Development of liver metastases is a frequent complication in patients with colorectal cancer (CRC), even after successful resection of the primary tumor. As such, post-operative adjuvant therapies that aim to eliminate residual disease after surgery may improve patient outcome.Methods: We used a colon carcinoma liver metastases model, in which CC531s colon carcinoma cells are injected into the portal circulation by a surgical procedure. As injected tumor cells are arrested in the liver, this model is suitable for investigating the interaction of tumor cells with the liver microenvironment. By administering tumor specific monoclonal antibodies (mAb) directly post-operatively, we were able to determine the effect of antibody therapy on eradication of arrested tumor cells and subsequent liver metastases outgrowth.Results: We showed that post-operative treatment with tumor specific monoclonal antibodies (mAb) prevents liver metastases outgrowth. Antibody-dependent phagocytosis (ADPh) was the main mechanism involved, as enhanced uptake of tumor cells by innate mononuclear phagocytes in the liver was observed after mAb therapy. Furthermore, Kupffer cells (KC) were identified as the most prominent effector cells, as depletion of KC abolished therapeutic efficacy. This was partly compensated by monocytes when animals were treated with a high mAb dose, but monocytes were unable to phagocytose tumor cells when rats were treated with low mAb doses.Conclusions: The finding that KC and monocytes can eliminate tumor cells through ADPh has important and promising clinical implications for designing new adjuvant therapies for patients undergoing CRC resection.

TRβ is the critical thyroid hormone receptor isoform in T3-induced proliferation of hepatocytes and pancreatic acinar cells

2010-07-05

Background & Aims: Thyroid hormones elicit many cellular and metabolic effects in various organs. Most of these actions, including mitogenesis, are mediated by the thyroid hormone 3,5,3′-triiodo-l-thyronine (T3) nuclear receptors (TRs). They are transcription factors, expressed as different isoforms encoded by the TRα and TRβ genes. Here, experiments were performed to determine whether (i) T3-induces hepatocyte proliferation in mouse liver and pancreas, and, (ii) which TR isoform, is responsible for its mitogenic effect.Methods: Cell proliferation was measured by bromodeoxyuridine (BrdU) incorporation after T3 or the TRβ agonist GC-1 in liver and pancreas of CD-1, C57BL, or TRα0/0 mice. Cell cycle-associated proteins were measured by Western blot.Results: T3 added to the diet at a concentration of 4mg/kg caused a striking increase in BrdU incorporation in mouse hepatocytes. Increased BrdU incorporation was associated with enhanced protein levels of cyclin D1 and PCNA and decreased levels of p27. Treatment with GC-1, a selective agonist of the TRβ isoform, also induced a strong mitogenic response of mouse hepatocytes and pancreatic acinar cells which was similar to that elicited by T3. Finally, treatment with T3 of mice TRα0/0 induced a proliferative response in the liver and pancreas, similar to that of their wild type counterpart.Conclusions: These results demonstrate that T3 is a powerful inducer of cell proliferation in mouse liver and suggest that the β-isoform is responsible for the hepatomitogenic activity of T3. The same isoform seems to also mediate the proliferation of mouse pancreatic acinar cells.

Hepatocytes express functional NOD1 and NOD2 receptors: A role for NOD1 in hepatocyte CC and CXC chemokine production

Melanie J. Scott, Christine Chen, Qian Sun, Timothy R. Billiar — 2010-06-23

Background & Aims: NOD-like receptors are recently described cytosolic pattern recognition receptors. NOD1 and NOD2 are members of this family that recognize bacterial cell wall components, diaminopimelic acid and muramyl dipeptide, respectively. Both NOD1 and NOD2 have been associated with many inflammatory diseases, although their role in liver inflammation and infection has not been well studied.Methods: We investigated the role of NOD receptors in mouse liver by assessing expression and activation of NOD1 and NOD2 in liver and primary isolated hepatocytes from C57BL/6 mice.Results: Both NOD1 and NOD2 mRNA and protein were highly expressed in hepatocytes and liver. RIP2, the main signaling partner for NODs, was also expressed. Stimulation of hepatocytes with NOD1 ligand (C12-iEDAP) induced NFκB activation, activation of MAP kinases and expression of chemokines CCL5 (RANTES) and CXCL1 (KC). C12-iEDAP also synergized with interferon (IFN)γ to increase iNOS expression and production of nitric oxide. Despite activating NFκB, NOD1 ligand did not upregulate hepatocyte production of the acute phase proteins lipopolysaccharide binding protein, serum amyloid A, or soluble CD14 in cell culture supernatants, or upregulate mRNA expression of lipopolysaccharide binding protein, serum amyloid A, C-reactive protein, or serum amyloid P. NOD2 ligand (MDP) did not activate hepatocytes when given alone, but did synergize with Toll-like receptor ligands, lipopolysaccharide (LPS), and polyI:C to activate NFκB and MAPK.Conclusions: All together these data suggest an important role for hepatocyte NOD1 in attracting leukocytes to the liver during infection and for hepatic NLRs to augment innate immune responses to pathogens.

Atorvastatin attenuates hepatic fibrosis in rats after bile duct ligation via decreased turnover of hepatic stellate cells

2010-07-01

Background & Aims: Activation of hepatic stellate cells (HSC) and transdifferentiation to myofibroblasts following liver injury is the main culprit for hepatic fibrosis. Myofibroblasts show increased proliferation, migration, contraction, and production of extracellular matrix (ECM). In vitro, HMG-CoA reductase inhibitors (statins) inhibit proliferation and induce apoptosis of myofibroblastic HSC. To investigate the antifibrotic effects of atorvastatin in vivo we used bile duct ligated rats (BDL).Methods: BDL rats were treated with atorvastatin (15mg/kg/d) immediately after ligation (prophylactically) or in on-going fibrosis (therapeutically). Fibrosis was assessed by hydroxyproline content and Sirius-red staining. The activation of HSC was investigated by analysis of αSMA expression. mRNA levels of cytokines and procollagen were analyzed by RT-PCR, and MMP-2 activity by zymography. Proliferation was assessed by expression of cathepsins (B and D), proliferating cell nuclear antigen (PCNA), and Ki67-staining. Apoptosis was characterized by caspase-3 activity, cleavage of PARP-1, and TUNEL assay. Hepatic inflammation was investigated by serum parameters and liver histology.Results: Prophylactic and early therapy with atorvastatin significantly attenuated fibrosis and HSC activation. Later therapy lacked significant effects on fibrosis but reduced profibrotic cytokine expression and led to a more quiescent state of HSC with less proliferation and apoptosis, while hepatic inflammation did not change.Conclusions: This study shows that very early atorvastatin treatment inhibits HSC activation and fibrosis in the BDL model in vivo, while late treatment reduces HSC turnover and activity. Our findings underline that long-term studies in humans are warranted.

Prevalence of non-alcoholic fatty liver disease and its association with cardiovascular disease in patients with type 1 diabetes

2010-06-29

Background & Aims: To estimate the prevalence of non-alcoholic fatty liver disease (NAFLD) in type 1 diabetic individuals, and to evaluate whether NAFLD is associated with increased prevalence of cardiovascular disease (CVD).Methods: All patients with diagnosed type 1 diabetes with available liver ultrasound data (n=250), who regularly attended our diabetes clinic, were enrolled. Main study measures were detection of NAFLD (by patient history and liver ultrasound) and asymptomatic/symptomatic CVD (by patient history, chart review, electrocardiogram, and echo-Doppler scanning of carotid and lower limb arteries).Results: The prevalence of NAFLD was 44.4%, and NAFLD was the most common cause (69.8%) of hepatic steatosis on ultrasound examination. Patients with NAFLD had a remarkably higher (p<0.001) age- and sex-adjusted prevalence of coronary (10.8% vs. 1.1%), cerebrovascular (37.3% vs. 5.5%) and peripheral (24.5% vs. 2.5%) vascular disease than their counterparts without NAFLD. In logistic regression analysis, NAFLD was associated with prevalent CVD (as composite endpoint), independently of age, sex, diabetes duration, hemoglobin A1c, smoking history, systolic blood pressure, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, triglycerides, and medication use (adjusted odds ratio 7.36, 95% confidence intervals 1.60–34.3, p<0.01).Conclusions: Our findings suggest that NAFLD is very common in type 1 diabetic subjects and is associated, independently of several confounding factors, with a higher prevalence of CVD. Future prospective studies are needed to evaluate whether NAFLD predicts incident CVD events in type 1 diabetes.

Hepatocellular ballooning in NASH

2010-07-01

Background & Aims: Hepatocellular ballooning is a key finding in nonalcoholic steatohepatitis (NASH). It is conventionally defined by hemotoxylin and eosin (H&E) staining showing enlarged cells with rarefied cytoplasm and recently by changes in the cytoskeleton. Fat droplets are emerging as important organelles in cell metabolism. To address a possible relation between fat droplets and ballooning, we studied fat staining, H&E, and keratin 18 staining in human NASH.Methods: Sequential staining and high resolution imaging were used to study freshly prepared cryo-sections from 10 patients with histologically confirmed steatohepatitis using oil red O for fat droplet identification, H&E to identify ballooning, and anti-K18 to confirm cytoskeletal changes. High resolution images were captured at each stage using the Aperio Scanscope. To provide ultrastructural correlation, glutaraldehyde-fixed specimens were studied using transmission electron microscopy (TEM) with serial sectioning for localization of ballooned cells by light microscopy and TEM in identical specimens.Results: Serial staining consistently demonstrated that hepatocellular ballooning is associated with fat droplet accumulation evident by oil red O positivity and depletion of cytoplasmic keratin 18 with K-18 positive Mallory–Denk bodies (MDB). TEM confirmed the association between osmium stained fat droplets, MDB formation, and cellular enlargement and suggested droplet-associated dilation of the endoplasmic reticulum.Conclusions: These results indicate a relationship between cellular ballooning, fat droplet accumulation, and cytoskeletal injury in NASH. We speculate that injury to multiple, organelles including fat droplets and endoplasmic reticulum, contribute to this characteristic finding.

Down-regulation of SREBP-1c is associated with the development of burned-out NASH

2010-07-08

Background & Aims: It is well-known that hepatic triglycerides (TG) diminish with the progression of non-alcoholic steatohepatitis (NASH), which has been designated as burned-out NASH, but its mechanism remains unclear. We aimed to explore the changes in hepatic fatty acid (FA) and TG metabolism with disease progression.Methods: Hepatic expression of key genes in healthy individuals (n=6) and patients with simple steatosis (SS, n=10), mild NASH (fibrosis stage 1–2, n=20), and advanced NASH (fibrosis stage 3–4, n=20) were assessed by quantitative polymerase chain reaction.Results: Hepatic expression of genes related to FA uptake and oxidation and very-low-density lipoprotein synthesis/export did not differ among the groups. However, the mRNA levels of sterol regulatory element-binding protein (SREBP)-1c and its downstream genes FA synthase, acetyl-coenzyme A carboxylase 1, and diacylglycerol acyltransferase 1 were inversely correlated with fibrosis stage. Immunoblot analysis revealed a remarkable reduction in mature SREBP-1c levels in advanced NASH. Furthermore, hepatic expression of tumor necrosis factor-α increased in accordance with fibrosis progression, which was possibly related to the decrease in hepatic SREBP-1c expression.Conclusions: Down-regulation of SREBP-1c and lipogenic enzymes may be associated with the development of burned-out NASH.

In vivo 3T spectroscopic quantification of liver fat content in nonalcoholic fatty liver disease: Correlation with biochemical method and morphometry

2010-06-16

Background & Aims: The clinical application of liver fat quantification has increased in recent years, paralleling the epidemic increase in nonalcoholic fatty liver disease. The aim of this study was to perform a diagnostic evaluation of spectroscopy by comparing its measurement of total lipid content with that from liver biopsies and morphometry in normal subjects and patients with nonalcoholic fatty liver disease.Methods: Patients with symptomatic cholelithiasis underwent 3T MR cholangiography with spectroscopic quantification of TLC. A laparoscopic cholecystectomy was performed on the day of admission, with liver samples taken during surgery. Microcolorimetric assessment quantified lipid content in liver samples and morphometric evaluation in stained slides. Statistical analysis included bivariate correlation, regression, and ROC analysis.Results: The study was conducted in 18 patients, 5 men (mean age, 35.2±11.03years; range, 27–54years) and 13 women (mean age, 46.77±11.77years; range, 21–61years). Using a cut-off value >5% for fat content, 8 patients presented with steatosis and 10 patients presented with normal liver fat content. A significant correlation was observed between fat spectroscopy and lipid content (r=0.876, p<0.001). A lower and non-significant correlation was observed between lipid content and morphometry (r=0.190, p>0.05).Conclusions: The accuracy of spectroscopy in assessing fat concentration with a cut-off level of 7.48% was 100%. Spectroscopy showed a strong and significant correlation with lipid content. It may reliably replace liver biopsy for the assessment of liver fat content.

Induced pluripotent stem cells: A new era for hepatology

2010-06-29

Stem cell transplantation has been proposed as an attractive alternative approach to restore liver mass and function. Recent progress has been reported on the generation of induced pluripotent stem (iPS) cells from somatic cells. Human-iPS cells can be differentiated towards the hepatic lineage which presents possibilities for improving research on diseases, drug development, tissue engineering, the development of bio-artificial livers, and a foundation for producing autologous cell therapies that would avoid immune rejection and enable correction of gene defects prior to cell transplantation. This focused review will discuss how human iPS cell advances are likely to have an impact on hepatology.

Hepatitis C virus infection: A “liaison a trois” amongst the virus, the host, and chronic low-level inflammation for human survival

Vincenzo Barnaba — 2010-07-02

This review covers the various aspects of the immune system that allows the relationship between the hepatitis-C virus, the host and chronic low-level inflammation, to be highly flexible and able to defend the host from persistent infections. This ambiguity mainly stems from the property of the immune system that can be both protective and harmful. Immunity cannot be fully protective without producing a certain degree of damage (acute hepatitis resulting in resolving HCV infection). In addition, the balance between protection and tissue damage is critical for the development of chronic HCV infection. The establishment of a state of chronic low-level inflammation is instrumental to limit liver immunopathology, to limit viral spread, and ultimately to ensure a long-lasting survival of the host. It is dictated by a fine equilibrium maintained by multiple immunologic mechanisms, including: sensory perception of innate immunity, virus-specific T and B cell functions, control of immune responses, and finally the balance between immunity and immunopathology that has principally evolved to favor the survival of the species.

Revising consensus in portal hypertension: Report of the Baveno V consensus workshop on methodology of diagnosis and therapy in portal hypertension

Roberto de Franchis, On behalf of the Baveno V Faculty — 2010-07-06

Portal hypertension is associated with the most severe complications of cirrhosis, including ascites, hepatic encephalopathy, and bleeding from gastro-esophageal varices. Despite the progress achieved over the last decades, the 6-week mortality associated with variceal bleeding is still in the order of 10–20%. Awareness of the difficulty inherent to the evaluation of diagnostic tools and the design and conduct of good clinical trials for the treatment of portal hypertension has led to the organization, since 1986, of a series of consensus meetings. The first one was organized by Andrew Burroughs in Groningen, The Netherlands . After Groningen, other meetings followed, in Baveno in 1990 (Baveno I) and in 1995 (Baveno II) , in Milan in 1992 , in Reston, USA, in 1996 , in Stresa in 2000 (Baveno III) , again in Baveno in 2005 (Baveno IV) , and in Atlanta in 2007 .

Barriers to the successful treatment of liver disease by hepatocyte transplantation

2010-07-06

Management of patients with hepatic failure and liver-based metabolic disorders is complex and expensive. Hepatic failure results in impaired coagulation, altered consciousness and cerebral function, a heightened risk of multiple organ system failure, and sepsis . Such manifold problems are only treatable today and for the foreseeable future by transplantation. In fact, whole or auxiliary partial liver transplantation is often the only available treatment option for severe, even if transient, hepatic failure. Patients with life-threatening liver-based metabolic disorders similarly require organ transplantation even though their metabolic diseases are typically the result of a single enzyme deficiency, and the liver otherwise functions normally. For all of the benefits it may confer, liver transplantation is not an ideal therapy, even for severe hepatic failure. More than 17,000 patients currently await liver transplantation in the United States, a number that seriously underestimates the number of patients that need treatment , as it has been estimated that more than a million patients could benefit from transplantation . Unfortunately, use of whole liver transplantation to treat these disorders is limited by a severe shortage of donors and by the risks to the recipient associated with major surgery .

Radiofrequency ablation for the treatment of HCC – Maybe much more than simple tumor destruction?

Tim F. Greten, Firouzeh Korangy — 2010-06-29

COMMENTARY ON: Radiofrequency thermal ablation for hepatocellular carcinoma stimulates autologous NK-cell response. Zerbini A, Pilli M, Laccabue D, Pelosi G, Molinari A, Negri E, Cerioni S, Fagnoni F, Soliani P, Ferrari C, Missale G. Gastroenterology, 2010 May;138(5):1931–1942. Eup 2010 Jan 11. Copyright 2010. Abstract reprinted with permission from Elsevier Inc.www.ncbi.nlm.gov/pubmed/20060829Abstract: Background & Aims: Radiofrequency thermal ablation (RFA) is a minimally invasive technique used as standard local therapy of hepatocellular carcinoma and second-line treatment for metastatic liver tumors. Studies in preclinical models and in patients have shown that thermal destruction of tumor tissue can enhance anti-tumor cellular responses, but our knowledge of its impact on natural killer (NK) cells is still very limited.Methods: Thirty-seven patients undergoing RFA for hepatocellular carcinoma were studied for peripheral blood lymphocytes counts followed by phenotypic and functional characterization of NK-cell population.Results: Peripheral blood lymphocytes kinetics revealed an increased frequency and absolute number of NK-cells expressing higher levels of activatory along with reduced levels of inhibitory NK receptors, and increased functional NK-cell activity. A prevalent expansion of the CD3(−)CD56(dim) NK subset was observed compared to the CD3(−)CD56(bright) counterpart. Interferon-gamma production, anti-K562 cell-cytotoxicity, and antibody-dependent cell-cytotoxicity, appeared consistently increased in terms of both absolute activity and killing efficiency at 4weeks after RFA, as compared to baseline. Interestingly, when recurrence-free survival was assessed in two groups of patients separated according to higher vs lower enhancement of cytotoxicity and/or interferon-gamma production, a significant difference was observed, thus suggesting a potential predictive role of NK functional assays on efficacy of RFA.Conclusions: RFA can lead to stimulation of NK-cells with a more differentiated and proactivatory phenotypic profile with general increase of functional activities. This observation may be relevant for development of adjuvant immunotherapeutic strategies aimed at enhancing NK-cell responses against primary and metastatic liver tumors. Copyright 2010 AGA Institute.

Building a bridge between obesity, inflammation and liver carcinogenesis

Jean-Charles Nault, Jessica Zucman-Rossi — 2010-07-02

COMMENTARY ON:: Dietary and genetic obesity promote liver inflammation and tumorigenesis by enhancing IL-6 and TNF expression. Eek Joong Park, Jun Hee Lee, Guann-Yi Yu, Guobin He, Syed Raza Ali, Ryan G. Holzer, Christoph H. Österreicher, Hiroyuki Takahashi and Michael Karin. Cell, January 2010;140(2):197–208. Copyright 2010. Abstract reprinted with permission from Elsevier Inc.www.ncbi.nlm.nih.gov/pubmed/20141834Abstract: Epidemiological studies indicate that overweight and obesity are associated with increased cancer risk. To study how obesity augments cancer risk and development, we focused on hepatocellular carcinoma (HCC), the common form of liver cancer whose occurrence and progression are the most strongly affected by obesity among all cancers. We now demonstrate that either dietary or genetic obesity is a potent bona fide liver tumor promoter in mice. Obesity-promoted HCC development was dependent on enhanced production of the tumor-promoting cytokines IL-6 and TNF, which cause hepatic inflammation and activation of the oncogenic transcription factor STAT3. The chronic inflammatory response caused by obesity and enhanced production of IL-6 and TNF may also increase the risk of other cancers.

HBV reactivation after fludarabine chemotherapy identified on investigation of suspected transfusion-transmitted Hepatitis B virus

2010-07-05

Background & Aims: Multi-transfused patients often receive treatments inducing various levels of immunodeficiency. Acute viral infections may then be attributed either to transfusion-transmitted infection (TTI) or reactivation of a past infection.Methods: A patient with chronic lymphocytic leukemia (CLL) who had >250 blood donor exposures developed acute Hepatitis B virus (HBV) infection. Routine donor testing for HB core antibodies (anti-HBc) was in place in the relevant period and investigations undertaken on the blood donors were negative.Results: Review of historical, molecular, and antigenic evidence demonstrated reactivation of a recovered HBV infection dating >30years and the selection of a rare escape mutant that briefly replicated and caused acute liver disease. This mutant was unreactive with several HBsAg assays and poorly reactive with an HBV vaccine plasma. Correcting the C139Y substitution by site directed mutagenesis of recombinant surface proteins re-established assay reactivity.Conclusions: Fludarabine, but not Chlorambucil, appeared sufficiently immunosuppressive to trigger reactivation despite low levels of neutralizing antibodies. Differentiating between TTI and reactivation of HBV becomes more challenging with the increasing frequency of immunocompromised blood recipients. Chemotherapy with Fludarabine alone should be considered as carrying high risk of viral reactivation. Pre-treatment testing and peripheral blood sample archiving may be indicated in HBsAg negative patients.

Journal of Hepatology

Editorial Board

Double announcement:Consensus Conference/Basic school

Monothematic Conference, Athens, Greece

Fellowship programme

International Liver Congress, Berlin, Germany

Contents

Focus

Prognosis indicator in acute liver failure: Is there a place for cell death markers?

Exploiting liver immunity for the prevention of hepatic metastases

A phase I clinical trial of dendritic cell immunotherapy in HCV-infected individuals

The novel immunoregulatory molecule FGL2: A potential biomarker for severity of chronic hepatitis C virus infection

Low virological response and high relapse rates in hepatitis C genotype 1 patients with advanced fibrosis despite adequate therapeutic dosing

Measurement of hepatic vein pressure gradient in children with chronic liver diseases

Prediction of oesophageal varices in hepatic cirrhosis by simple serum non-invasive markers: Results of a multicenter, large-scale study

Cytokeratin 18-based modification of the MELD score improves prediction of spontaneous survival after acute liver injury

Activation of human liver sinusoidal endothelial cell by human platelets induces hepatocyte proliferation

Early growth response-1 attenuates liver injury and promotes hepatoprotection after carbon tetrachloride exposure in mice

Pancreatic Duodenal Homeobox-1 de novo expression drives cholangiocyte neuroendocrine-like transdifferentiation

Genetic variations of hepatitis B virus and serum aflatoxin-lysine adduct on high risk of hepatocellular carcinoma in Southern Guangxi, China

Experimentally induced liver metastases from colorectal cancer can be prevented by mononuclear phagocyte-mediated monoclonal antibody therapy

TRβ is the critical thyroid hormone receptor isoform in T3-induced proliferation of hepatocytes and pancreatic acinar cells

Hepatocytes express functional NOD1 and NOD2 receptors: A role for NOD1 in hepatocyte CC and CXC chemokine production

Atorvastatin attenuates hepatic fibrosis in rats after bile duct ligation via decreased turnover of hepatic stellate cells

Prevalence of non-alcoholic fatty liver disease and its association with cardiovascular disease in patients with type 1 diabetes

Hepatocellular ballooning in NASH

Down-regulation of SREBP-1c is associated with the development of burned-out NASH

In vivo 3T spectroscopic quantification of liver fat content in nonalcoholic fatty liver disease: Correlation with biochemical method and morphometry

Induced pluripotent stem cells: A new era for hepatology

Hepatitis C virus infection: A “liaison a trois” amongst the virus, the host, and chronic low-level inflammation for human survival

Revising consensus in portal hypertension: Report of the Baveno V consensus workshop on methodology of diagnosis and therapy in portal hypertension

Barriers to the successful treatment of liver disease by hepatocyte transplantation

Radiofrequency ablation for the treatment of HCC – Maybe much more than simple tumor destruction?

Building a bridge between obesity, inflammation and liver carcinogenesis

HBV reactivation after fludarabine chemotherapy identified on investigation of suspected transfusion-transmitted Hepatitis B virus