Fibrosis In Chronic Hepatitis C Research Articles

Cytokine profiling of plasma in patients with viral hepatitis C

Chronic hepatitis C (CHC) represents a significant public health concern. In the majority of cases, the infection progresses to a chronic form, which is characterised by the development of fibrosis and cirrhosis of the liver. A plethora of cytokines and chemokines are generated as a consequence of inflammatory processes within the liver. These can exert a dual effect, both protective and damaging, particularly in relation to the death of hepatocytes and the progression of liver fibrosis. Furthermore, a number of growth factors have been identified as playing a role in the pathogenesis of CHC. The objective of the study was a comprehensive evaluation of a wide range of cytokines, chemokines and growth factors in the blood plasma of patients with CHC at varying stages of liver fibrosis. The study cohort comprised 63 patients diagnosed with CHC, who were divided into three groups according to the stage of liver fibrosis. The control group comprised healthy individuals (n = 32). Concentrations of the following cytokines were determined in plasma: Interleukins and some cytokines (IL-1α, IL-1β, IL-1ra, IL-2, IL-4, IL-5, IL-6, IL-7, IL-9, IL-10, IL-12 (p40), IL-12 (p70), IL-13, IL-15, IL-17A, IL-17-E/IL-25, IL-17F, IL-18, IL-27, IFNα, IFNγ, TNFα, TNFβ); chemokines (CCL2/MCP-1, CCL3/MIP-1α, CCL4/MIP-1β, CCL7/MCP-3, CCL11/Eotaxin, CCL22/MDC, CXCL1/GROα, CXCL8/IL-8, CXCL9/MIG, CXCL10/IP-10, CX3CL1/Fractalkine) and growth factors (EGF, FGF-2, Flt-3L, G-CSF, M-CSF, PDGF-AA, PDGF-AB/BB, TGF-α, VEGF-A) by multiplex analysis based on xMAP technology. Nonparametric statistics methods were used for statistical analysis. As a result of the study, increased concentrations of cytokines IL-12 (p40), IL-15, IL-17E/IL-25, IL-27, IFNγ, TNFα, chemokines CXCL9/MIG and CXCL-10/IP-10 and growth factors FGF-2 and M-CSF were found at all stages of liver fibrosis. Elevated concentrations of cytokines IL-1α, IL-1β, IL-2, IL-6, IL-9, IL-10, IL-17F, IFNα, TNFβ, chemokines CCL2/MCP-1, CCL11/Eotaxin, CCL22/MDC and growth factors G-CSF, TGF-α, Flt-3L were found in severe liver fibrosis/cirrhosis. Correlation analysis revealed a relationship of high significance between the severity of liver fibrosis and the content of cytokines IL-6, IFNγ, TNFα, IL-7, chemokines CCL2/MCP-1, CCL11/Eotaxin, CXCL9/MIG, CXCL10/IP-10, CXCL1/GROα, growth factors TGF-α, PDGF-AA, PDGF-AB/BB. Thus, a certain profile of cytokines characteristic for CHC was revealed, cytokines, chemokines and growth factors significant for liver fibrosis in CHC were found.

ASPARTATE PLATELET RATIO INDEX AS A PREDICTOR OF SEVERITY OF FIBROSIS IN CHRONIC HEPATITS C

Introduction: Hepatitis C is an important emerging cause for chronic liver disease in India, with high risk for chronicity and hepatocellular carcinoma. Chronic hepatitis C (CHC) is the most common cause for chronic liver disease and cirrhosis, and liver transplantation. Liver biopsy is the gold standard for evaluation of fibrosis - however it remains fraught with drawbacks and limitations. The aspartate aminotransferase-to-platelet ratio index (APRI), a tool with limited expense and widespread availability is a promising non-invasive alternative to liver biopsy for detecting hepatic fibrosis in CHC. Objectives: (1) The objectives of the study were to determine the association between APRI and severity of fibrosis in Hepatitis C (2) and to describe the clinical profile of patients with Hepatitis C. Methods: A cross-sectional descriptive study in 60 patients diagnosed with hepatitis C for more than 6 months admitted to the General Medicine and Medical Gastroenterology Departments of Govt Medical College, Kottayam. Data were collected with a structured pro forma and analyzed using SPSS. Results: 35 of 60 patients belonged to METAVIR F3 (severe fibrosis) and F4 (cirrhosis). An APRI of 0.5 was associated with a finding of F3 or F4 with a sensitivity of 97.14% and a specificity of 88%. 31 of the 60 patients belonged to F4. An APRI of 1.5 or more was a predictor for cirrhosis with a sensitivity of 93.1% and specificity of 96.77%. The positive predictive value of cirrhosis APRI threshold >1.5 is 96.4%.

ХАРАКТЕРИСТИКА ФЕНОТИПОВ СТЕЛЛАТНЫХ КЛЕТОК ПЕЧЕНИ ПРИ ХРОНИЧЕСКОМ ГЕПАТИТЕ С

Background. Hepatic stellate cells (HSC) play a key role in the development of liver fibrosis in different damages. The aim is to present the structural-functional and quantitative characteristics of various HSC phenotypes in chronic hepatitis C (CHC). Material and methods. The object of the study was 18 liver biopsies of patients with verified CHC (HCV+ RNA in PCR). The stage of liver fibrosis was assessed by Metavir. Structural, functional and quantitative characteristics of different phenotypes of 160 analyzed HSCs at different stages of fibrosis in CHC were assessed by the results of light (semithin sections) and electron microscopy. We determined the average area, length and width of one PSL in each biopsy specimen, the number of lipid droplets in all and in one PSL as well as the average area, length and width of one lipid droplet. Results. At different stages of fibrosis in CHC, all three HSC phenotypes, which had differences in the main structural and quantitative parameters, were simultaneously detected in the patient. Predominantly (44.4%), HSCs corresponding to the non-activated (sleeping) phenotype were detected, less often (20.6%) – those corresponding to the active (myofibroblastic) phenotype. As the HSC was activated, the shape of the cells changed (star-shaped–elongated), the number of lipid droplets in the cell decreased without changing the size of the inclusions, and the cytoplasmic/nuclear ratio changed towards an increase in the nucleus. The frequency distribution of phenotypes depended on the stage of liver fibrosis. Conclusions. Patients with CHC are characterized by the presence of all three HSC phenotypes, which differ from each other in structural and quantitative characteristics, the frequency of which depends on the stage of liver fibrosis.

Intercellular interactions and their role in fibrogenesis in chronic hepatitis C

Introduction. The development of life-threatening complications in chronic hepatitis C (CHC) is based on progressive fibrogenesis. The developing of liver fibrosis is provided by intercellular interactions, first of all, of lymphocytes, macrophages and stellate cells (SC), the patterns of mutual influences of which have not been sufficiently studied at the moment.The objective was to study the features of intercellular interplay of nonparenchymal liver cells at different histological activity, at different stages of CHC fibrosis, and at different genotypes of the hepatitis C virus (HCV).Methods and materials. The object of the study was 64 liver biopsies of adult patients with natural course of CHC. Нistological, immunohistochemical and immunohistomorphometric methods were used.Results. The increasing histological activity is accompanied by an increase in the number and size (area) of CD68 + macrophages and SMA-alfa + SC. Correlation relationships of intercellular interactions at low and moderate histological activity had significant differences. In mild fibrosis, a relationship was found between the number of CD8 + lymphocytes, the number and area of CD68 + macrophages and SMA-alfa + SC. HCV genotype 1 is characterized by a predominance of the interactions between the number of CD8 + lymphocytes, the number and area of CD68 + macrophages in the liver, for genotype 3 – between the number of CD8 + lymphocytes, the number and area of SMA + SC.Conclusions. The maximum activation of SC and macrophages occurs even with moderate histological activity and persists with an increase. The consolidation of the immunopathological nature of the intercellular interplay between lymphocytes, macrophages and SC occurs at the stage of mild fibrosis. Intercellular interactions have significant differences depending on the HCV genotype, which can determine a poor prognosis of the disease.

<p>The Utility of Retinol-Binding Protein 4 in Predicting Liver Fibrosis in Chronic Hepatitis C Patients in Response to Direct-Acting Antivirals</p>

BackgroundHepatic fibrosis grading is crucial for chronic hepatitis C (CHC) patients in monitoring liver disease progression and antiviral treatment indication. Retinol-binding protein 4 (RBP4), an adipokine secreted by adipocytes and hepatocytes, has variable levels in health and disease.PurposeTo comparatively evaluate RBP4 serum levels in predicting liver fibrosis in CHC versus fibroscan, noninvasive fibrosis, and inflammatory indices.Patients and MethodsCohort study included 50 naive non-obese CHC patients and 20 age-, sex- and body mass index-matched healthy subjects. Fibroscan, RBP4, and noninvasive fibrosis as APRI, CDS, FIB-4, GUCI, Lok index indices based on serological markers, and inflammatory indices as platelet to lymphocyte ratio (PLR) and liver regeneration markers as; alpha-fetoprotein (AFP) and APRI, were evaluated in response to direct-acting antivirals (DAAs).ResultsRBP4 was significantly lower in patients than in controls (P=0.0001) and progressively decreased with the increase in fibrosis grade (F0-F=41.42±3.08), (F2=39.32±1.43), (F3-F4= 35.31±0.5), (P=0.0001). Liver function, stiffness, and RBP4 significantly improved after treatment (P=0.0001). RBP4 negatively correlated with viral load (r=−0.78, p=0.0001), fibroscan fibrosis grade (r=−0.52, p=0.0001), AFP (r=−0.63, p=0.0001), and positively correlated with platelet (r=0.424, p=0.0001), and white cell count (r=0.298, p=0.002). RBP4 at a cutoff value <40.55 ng/mL might predict significant fibrosis (90.48% sensitivity, 62.5% specificity, AUROC=0.811, 95% CI=67.5–90.0) and at a cutoff value <35.9 ng/mL could predict advanced fibrosis (100% sensitivity, 100% specificity, AUROC =1.0, 95% CI=0.929–1).ConclusionRBP4 showed excellent accuracy, sensitivity, specificity, PPV, and NPV. RBP4 has a superior diagnostic performance in predicting advanced fibrosis grads in CHC patients and hence can replace expensive invasive procedures.

A study of serum miRNA-122 in hepatitis C and associated hepatocellular carcinoma

BACKGROUND: The discovery of a cluster of short non-coding RNAs called microRNAs (miRNAs) has become an important event in molecular biology. One of its representatives, miR-122 plays a large role in regulating the expression of genes involved in carbohydrate, lipid metabolism, and iron metabolism in the body. In experimental studies it was shown that in addition to regulatory functions, miR-122 is involved in the pathogenesis of hepatitis C, providing the life cycle of the virus in the cell. The shift of emphasis in the study of miR-122 from basic research into clinical medicine seems to be a promising area of personalized medicine.&#x0D; AIMS: to determine the clinical significance of miR-122 in acute and chronic hepatitis C and associated hepatocellular carcinoma.&#x0D; MATERIALS AND METHODS: A total of 407 people were examined, including 17 patients with acute hepatitis C (AHC), 158 patients with chronic hepatitis C (CHC) and 62 patients with HCC associated with hepatitis C. Comparison groups consisted of 84 healthy individuals and 62 patients with clinically pronounced cirrhosis of a non-infectious etiology. In each cohort, the relative miR-122 level was determined in the blood of patients. The analysis was performed in PCR using the Qubit microRNA Assay Kit -100 for the quantitative determination of microRNAs (Thermo Fisher Scientific, USA). Relative miR-122 expression values were calculated by the formula 2 -CT using U6 snRNA as a reference RNA.&#x0D; RESULTS: The highest miR-122 level in serum was found in patients with AHC at the height of the icteric period. The level of miR-122 showed a direct correlation with the activity of hepatic transaminases in patients with AHC (r = 0.72) and CHC (r = 0.44). An analysis of miR-122 level relative to the degree of liver fibrosis in patients with chronic hepatitis C showed that, as liver fibrosis progresses, the level of miR-122 expression decreases. The decrease in miR-122 expression in patients with severe fibrosis was universal and did not depend on the etiology of the disease. The development of HCC in the presence of chronic hepatitis C was accompanied by a decrease in the level of miR-122 by 10 times on average compared to patients with chronic hepatitis C.&#x0D; CONCLUSIONS: The determination of the expression level of miR-122 in serum can be used in laboratory monitoring of the management of patients with HC as an indicator of the severity of liver damage in AHC and the rate of formation of liver fibrosis in CHC. Evaluation of possibility of using miR-122 as a predictor of the development of HCC in the outcome of HC requires additional studies of the specificity and sensitivity of the test and comparison of the obtained data with the results of using generally accepted protein tumor markers.

Association between PNPLA3[G]/I148M variant, steatosis and fibrosis stage in hepatitis C virus - genetic matters.

There is an established correlation between the PNPLA3 rs738409 C > G single nucleotide polymorphism (SNP) and hepatic steatosis and fibrosis in hepatitis C virus (HCV) infected patients. However not all data is convergent regarding the exact impact of this SNP on the pattern of disease progression in different clinical settings. In this study, we aimed to further bridge the knowledge gap on this topic by investigating the role of the G allele in promoting steatosis, fibrosis and disease progression in relation to other metabolic and anthropometric host factors. Two hundred and fifty consecutive patients, previously diagnosed with chronic hepatitis C (CHC) underwent liver biopsy. Histology was assessed using the Metavir scoring system. Transient elastography was used for follow-up. Ninety-eight patients were genotyped for PNPLA3 rs738409 and followed up for fibrosis progression. PNPLA3 rs738409[G] allele was significantly correlated with severe steatosis (P = 0.04), severe fibrosis at the time of enrollment (P = 0.0005) and fibrosis progression with an OR of 10.31 (95% CI 1.06 - 99.59, P = 0.04), after a mean follow-up time of 62.85 (95%CI: 52.21 - 76.15) months. Severe steatosis at the time of enrollment had an OR of 11.02 (95% CI 1.48 - 82.09, P = 0.01) for the association with fibrosis progression. The HOMA-IR index was also positively correlated with severe fibrosis (P = 0.03) and fibrosis progression on univariate analysis (P = 0.02). PNPLA3 rs738409[G] allele is a reliable predictor for steatosis and fibrosis in CHC. The presence of G allele, along with severe steatosis and insulin resistance are significant predictors for fibrosis progression.

MicroRNA-10a Impairs Liver Metabolism in Hepatitis C Virus-Related Cirrhosis Through Deregulation of the Circadian Clock Gene Brain and Muscle Aryl Hydrocarbon Receptor Nuclear Translocator-Like 1.

The circadian rhythm of the liver plays an important role in maintaining its metabolic homeostasis. We performed comprehensive expression analysis of microRNAs (miRNAs) using TaqMan polymerase chain reaction of liver biopsy tissues to identify the miRNAs that are significantly up‐regulated in advanced chronic hepatitis C (CHC). We found miR‐10a regulated various liver metabolism genes and was markedly up‐regulated by hepatitis C virus infection and poor nutritional conditions. The expression of miR‐10a was rhythmic and down‐regulated the expression of the circadian rhythm gene brain and muscle aryl hydrocarbon receptor nuclear translocator‐like 1 (Bmal1) by directly suppressing the expression of RA receptor‐related orphan receptor alpha (RORA). Overexpression of miR‐10a in hepatocytes blunted circadian rhythm of Bmal1 and inhibited the expression of lipid synthesis genes (sterol regulatory element binding protein [SREBP]1, fatty acid synthase [FASN], and SREBP2), gluconeogenesis (peroxisome proliferator‐activated receptor gamma coactivator 1 alpha [PGC1α]), protein synthesis (mammalian target of rapamycin [mTOR] and ribosomal protein S6 kinase [S6K]) and bile acid synthesis (liver receptor homolog 1 [LRH1]). The expression of Bmal1 was significantly correlated with the expression of mitochondrial biogenesis‐related genes and reduced Bmal1 was associated with increased serum alanine aminotransferase levels and progression of liver fibrosis in CHC. Thus, impaired circadian rhythm expression of Bmal1 by miR‐10a disturbs metabolic adaptations, leading to liver damage, and is closely associated with the exacerbation of abnormal liver metabolism in patients with advanced CHC. In patients with hepatitis C‐related liver cirrhosis, liver tissue miR‐10a levels were significantly associated with hepatic reserve, fibrosis markers, esophageal varix complications, and hepatitis C‐related hepatocellular carcinoma recurrence. Conclusion: MiRNA‐10a is involved in abnormal liver metabolism in cirrhotic liver through down‐regulation of the expression of the circadian rhythm gene Bmal1. Therefore, miR‐10a is a possible useful biomarker for estimating the prognosis of liver cirrhosis.

PNPLA3 Polymorphism Directly Affects the Susceptibility of Genotype 4-Induced HCV Infection and Its Progression to Liver Fibrosis Among the Egyptian Patients

Chronic Hepatitis C (CHC) infection is associated with liver fibrosis, cirrhosis, hepatocellular carcinoma, and eventually death. Genetic risk factors have been casted as predictors of the severity and progression of several CHC-associated liver diseases. PNPLA3 (rs738409), a novel genetic marker strongly associated with increased fat content in the liver. Nonetheless, recognition of an association between PNPLA3 rs73409 (C>G) and the severity of fibrosis in CHC has been inadequately studied.

Circulating let-7 Levels in Serum Correlate With the Severity of Hepatic Fibrosis in Chronic Hepatitis C.

BackgroundEvaluating the progression of hepatic fibrosis in chronic hepatitis C (CHC) is critical, and identifying a predictive biomarker for fibrosis will be helpful for implementing personalized surveillance of hepatocellular carcinoma after the elimination of hepatitis C virus by antiviral therapy. This study aimed to investigate the association of circulating let-7a-5p levels with severity of hepatic fibrosis. MethodsWe analyzed circulating let-7a-5p levels in serum and serum-derived extracellular vesicles (EVs) in 84 Japanese CHC patients who underwent a liver biopsy by quantitative real-time polymerase chain reaction, and investigated the association of its levels with histological hepatic fibrotic stage, liver stiffness, and several hepatic fibrotic markers.ResultsThe levels of let-7a-5p in serum and EVs were significantly lower in patients with liver cirrhosis. Additionally, the serum let-7a-5p level correlated significantly with hepatic fibrotic markers, Mac-2 binding protein glycan isomer (M2BPGi), fibrosis-4 (FIB-4) index, aspartate aminotransferase-to-platelet ratio index (APRI), and liver stiffness, evaluated by transient elastography. Furthermore, the serum let-7a-5p level was superior to M2BPGi, FIB-4, and APRI and was comparable to liver stiffness in discriminating liver cirrhosis. ConclusionsThese results provide evidence that circulating let-7a-5p in serum may serve as a surrogate marker for severity of hepatic fibrosis in CHC.

Interferon Lambda 4 RS12979860 C>T polymorphism is not associated with liver fibrosis in patients with hepatitis C

Host genetic predispositions may be important determinants of liver fibrosis in patients with chronic hepatitis C (CHC). The association between Interferon-L 4 (IFNL4) rs12979860 C>T polymorphism and risk of liver fibrosis in CHC is contradictory. To evaluate the impact of IFNL4 rs12979860 polymorphism on the risk of fibrosis in patients with CHC. One hundred fifty patients with CHC aged 50 ± 11 years (89 females) were genotyped for IFNL4 rs12979860 using real time PCR. Fibrosis present in liver biopsies was assessed using the METAVIR score, comparing patients with either no fibrosis, mild fibrosis, or intermediate fibrosis (F0+F1+F2, n = 96), with patients with severe fibrosis or cirrhosis (F3+F4, n = 54). In F0-F2 patients the distribution of rs12979860 genotypes was 22 CC, 57 CT and 17 TT, whereas in patients F3-F4 the distribution was 10, 29 and 15, respectively. No association between IFNL4 rs12979860 genotype and risk of fibrosis was observed in uni or multivariate analyses. IFNL4 rs12979860 C>T polymorphism is not associated with risk of liver fibrosis in this group of patients with CHC.

Triplex High-Resolution Melting Assay for the Simultaneous Assessment of IFNL3 rs12979860, ABCB11 rs2287622, and RNF7 rs16851720 Genotypes in Chronic Hepatitis C Patients

Chronic hepatitis C (CHC) is a leading cause of liver disease. Despite the improved efficacy of new antivirals, their high costs preclude their adoption in resource-limited settings, where CHC prevalence is highest. We developed a triplex high-resolution melting assay for the simultaneous assessment of three genetic polymorphisms related to the response to treatment and development of advanced fibrosis in CHC: IFNL3 rs12979860, ABCB11 rs2287622, and RNF7 rs16851720. We validated the assay in clinical samples from 130 CHC patients treated with classic therapy. The assay showed excellent reproducibility and 100% accuracy, sensitivity, and specificity against the gold standard Sanger sequencing. When added to routine examination data, genotype information significantly improved their performance for prediction of advanced liver fibrosis and sustained virological response (P=0.041 and P = 0.011, respectively). Correspondingly, the full models had area under the receiver operating characteristic curve values of 0.842 (95% CI, 0.773-0.911) and 0.921 (95% CI, 0.870-0.972) and integrated discrimination improvements of 7.5% (95% CI, 2.5%-12.5%; P=0.003) and 11.5% (95% CI, 5.8%-17.2%; P<0.001), respectively. This is the first report on a diagnostic test for simultaneous genotyping of IFNL3, ABCB11, and RNF7 in CHC patients. Reliable and inexpensive, the assay should provide useful information for the clinical management of CHC, like identification of patients at risk of rapiddisease progression or with high chances of response to classic therapy.

Fibrosis in Chronic Hepatitis C: Correlation between Immunohistochemically-Assessed Virus Load with Steatosis and Cellular Iron Content.

AIM:We aimed study impact of hepatocytic viral load, steatosis, and iron load on fibrosis in chronic hepatitis C and role of VEGF and VEGFR overexpression in cirrhotic cases in evolving HCC.MATERIAL AND METHODS:Total of 120 cases were included from TBRI and Beaujon Hospital as chronic hepatitis C (CHC), post-hepatitis C cirrhosis, and HCC. Cases of CHC were stained for Sirius red, Prussian blue and immunohistochemically (IHC) for HCV-NS3/NS4. HCC were stained IHC for VEGF and by FISH.RESULTS:Stage of fibrosis was significantly correlated with inflammation in CHC (P < 0.01). Noticed iron load did not correlate with fibrosis. Steatosis was associated with higher inflammation and fibrosis. The cellular viral load did not correlate with inflammation, steatosis or fibrosis. VEGF by IHC was significantly higher in cases of HCC when compared to cirrhotic group (P < 0.001). Amplification of VEGFR2 was confirmed in 40% of cases of HCC. Scoring of VEGF by IHC was the good indicator of VEGFR2 amplification by FISH (P < 0.005).CONCLUSION:Grade of inflammation is the factor affecting fibrosis in CHC. The degree of liver damage is not related to cellular viral load or iron load. Steatosis is associated with higher inflammation and fibrosis. VEGF by IHC is correlated with overexpression of VEGFR2 by FISH.

Circulating let-7 levels in plasma and extracellular vesicles correlate with hepatic fibrosis progression in chronic hepatitis C.

The goal of this study was to determine whether an association exists between circulating microRNA (miRNA) levels and disease progression in chronic hepatitis C (CHC), whether plasma or extracellular vesicles (EVs) were optimal for miRNA measurement and their correlation with hepatic miRNA expression, and the mechanistic plausibility of this association. We studied 130 CHC patients prospectively followed over several decades. A comprehensive miRNA profile in plasma using microarray with 2578 probe sets showed 323 miRNAs differentially expressed between healthy individuals and CHC patients, but only six that distinguished patients with mild versus severe chronic hepatitis. Eventually, let-7a/7c/7d-5p and miR-122-5p were identified as candidate predictors of disease progression. Cross-sectional analyses at the time of initial liver biopsy showed that reduced levels of let-7a/7c/7d-5p (let-7s) in plasma were correlated with advanced histological hepatic fibrosis stage and other fibrotic markers, whereas miR-122-5p levels in plasma were positively correlated with inflammatory activity, but not fibrosis. Measuring let-7s levels in EVs was not superior to intact plasma for discriminating significant hepatic fibrosis. Longitudinal analyses in 60 patients with paired liver biopsies showed that let-7s levels in plasma markedly declined over time in parallel with fibrosis progression. However, circulating let-7s levels did not parallel those in the liver. Of all miRNAs screened, the let-7 family showed the best correlation with hepatic fibrosis in CHC. A single determination of let-7s levels in plasma did not have superior predictive value for significant hepatic fibrosis compared with that of fibrosis-4 index, but the rate of let-7s decline in paired longitudinal samples correlated well with fibrosis progression. Pathway analysis suggested that low levels of let-7 may influence hepatic fibrogenesis through activation of transforming growth factor β signaling in hepatic stellate cells. (Hepatology 2016;64:732-745).

Diverse impacts of the rs58542926 E167K variant in TM6SF2 on viral and metabolic liver disease phenotypes.

A genome-wide exome association study has identified the transmembrane 6 superfamily member 2 (TM6SF2) rs58542926 variant encoding an E167K substitution as a genetic determinant of hepatic steatosis in nonalcoholic fatty liver disease (NAFLD). The roles of this variant across a spectrum of liver diseases and pathologies and on serum lipids comparing viral hepatitis to NAFLD and viral load in chronic viral hepatitis, as well as its intrahepatic molecular signature, have not been well characterized. We undertook detailed analyses in 3260 subjects with viral and nonviral liver diseases and in healthy controls. Serum inflammatory markers and hepatic expression of TM6SF2 and genes regulating lipid metabolism were assessed in a subset with chronic hepatitis C (CHC). The rs58542926 T allele was more prevalent in 502 NAFLD patients than controls (P = 0.02) but not different in cohorts with CHC (n = 2023) and chronic hepatitis B (n = 507). The T allele was associated with alterations in serum lipids and hepatic steatosis in all diseases and with reduced hepatic TM6SF2 and microsomal triglyceride transfer protein expression. Interestingly, the substitution was associated with reduced CHC viral load but increased hepatitis B virus DNA. The rs58542926 T allele had no effect on inflammation, impacted ≥F2 fibrosis in CHC and NAFLD assessed cross-sectionally (odds ratio = 1.39, 95% confidence interval 1.04-1.87, and odds ratio = 1.62, 95% confidence interval 1.03-2.52, respectively; P < 0.03 for both), but had no effect on fibrosis progression in 1174 patients with CHC and a known duration of infection. The TM6SF2 E167K substitution promotes steatosis and lipid abnormalities in part by altering TM6SF2 and microsomal triglyceride transfer protein expression and differentially impacts CHC and chronic hepatitis B viral load, while effects on fibrosis are marginal. (Hepatology 2016;64:34-46).

Diagnostic Performance of Real-Time Elastography in the Assessment of Advanced Fibrosis in Chronic Hepatitis C

Background and aimsSince liver fibrosis index (LFI) was developed by Fujimoto et al., real-time elastography (RTE) has become a promising non-invasive technique to assess fibrosis in chronic hepatitis C (CHC). The aims of this study were to compare the diagnostic performance of RTE versus laboratory tests to predict advanced fibrosis (METAVIR scoring system: F≥3) in patients with CHC, using liver biopsy (LB) as the reference standard; and to evaluated the impact of patient anthropometric features on RTE histogram acquisition. MethodsThis prospective study included 37 patients with CHC scheduled for LB. Aspartate aminotransferase (AST)/alanine aminotransferase (AST) ratio, AST/platelet ratio index (APRI), and Fibrosis-4 index (FIB-4) were calculated from recent (≤6 months) laboratory data. RTE was performed by two independent operators blind to each other’ findings and to LB results, using Hitachi HI-VISION Avius ultrasound system. According to Hitachi RTE software, liver elasticity was evaluated through the LFI. Percutaneous ultrasound-assisted LB was performed in the same day of RTE. All LB specimens were analyzed by an expert pathologist blind to RTE results. Hepatic fibrosis was staged according to METAVIR scoring system. The diagnostic performance of the LFI, AST/ALT ratio, APRI and FIB-4 for predicting advanced fibrosis was assessed using area under receiver-operating characteristic curve (AUROC), sensitivity, specificity, positive-predictive and negative-predictive (NPV) values. ResultsThirty-seven LB were performed without complications. The distribution according to METAVIR scoring system was F0–1 in 13 patients (35%), F2 in 13 (35%), F3 in 9 (25%) and F4 in 2 (5%). Thirty-seven RTE procedures were performed. Histogram acquisition was successfully achieved in 32 patients (86%). Abdominal wall thickness ≥23mm was associated with no histogram acquisition (p=0.018). Using the optimal cut-off value of 2.38, the AUROC for the LFI was 0.73. The AUROC for the AST/ALT ratio, APRI and FIB-4 were 0.62, 0.79, and 0.82, respectively. ConclusionsThe LFI calculated by RTE showed a very good diagnostic performance to predict advanced fibrosis in CHC, with remarkable sensitivity and NPV (both 100%).

PNPLA3 rs738409 Polymorphism Associated with Hepatic Steatosis and Advanced Fibrosis in Patients with Chronic Hepatitis C Virus: A Meta-Analysis.

Background/AimsThe recognition of a correlation between patatin-like phospholipase domain containing-protein 3 (PNPLA3) rs738409 (C>G) and the severity of liver steatosis or fibrosis in chronic hepatitis C (CHC) has not reached a consensus. This meta-analysis sought to investigate with accuracy the association between the PNPLA3 rs738409 (C>G) polymorphism and liver steatosis and advanced fibrosis in CHC patients.MethodsWe performed a comprehensive literature search from the PubMed, Embase, Web of Science, and Google Scholar databases up to December 31, 2014. Crude odds ratios (ORs) with 95% confidence intervals (CIs) were calculated. Statistical analyses were performed using Stata 12.0 software.ResultsThe meta-analysis revealed the severity of liver fibrosis was significantly higher in CHC patients with PNPLA3 rs738409 GG in Caucasians (versus CC+CG: OR, 2.29; 95% CI, 1.57 to 3.35; p<0.05) but not Asian populations. In Caucasians, liver steatosis was also more severe in CHC patients with rs738409 GG (versus CC+CG; OR, 4.33; 95% CI, 2.59 to 7.22; p<0.05). The sensitivity analysis indicated the results of this meta-analysis were stable and no publication bias was detected.ConclusionsPNPLA3 rs738409 (C>G) was associated with the risk of both advanced liver fibrosis and steatosis in patients with CHC, especially among Caucasian populations.

Utility of real-time shear wave elastography for assessing liver fibrosis in patients with chronic hepatitis C infection without cirrhosis: Comparison of liver fibrosis indices.

To clarify the diagnostic impact of liver fibrosis except for cirrhosis identified using shear wave elastography (SWE) in chronic hepatitis C (CHC) patients, and to compare the performance in diagnosing liver fibrosis among SWE and liver fibrosis indices. A total of 55 CHC patients who underwent liver biopsy were analyzed. The diagnostic performance for identifying significant liver fibrosis (F2-F3) for SWE, FIB-4 index, aspartate aminotransferase-to-platelet ratio index (APRI) and Forns' index was assessed using receiver-operator curve (ROC) analysis. The median SWE elasticity value, FIB-4 index, APRI and Forns' index in the F0-F1 and F2-F3 groups were 6.3 kPa and 13.1 kPa; 1.52 and 4.45; 0.41 and 1.43; and 7.69 and 8.85, respectively (P < 0.001 for all four methods). Multivariate analysis showed that SWE was independently associated with the presence of significant liver fibrosis (odds ratio, 2.52; 95% confidence interval, 1.49-4.28; P < 0.001). The area under the ROC curve for SWE in diagnosing significant liver fibrosis was 0.94, indicating high diagnostic value, compared with 0.86, 0.88 and 0.83, for the FIB-4 index, APRI and Forns' index, respectively, which corresponds to moderate diagnostic value. The accuracy of SWE, FIB-4 index, APRI and Forns' index for diagnosing significant liver fibrosis was 90.9%, 76.4%, 74.5% and 67.2%, respectively. SWE has excellent ability for diagnosing significant liver fibrosis in CHC even when patients with cirrhosis are excluded. The diagnostic performance of SWE is superior to that of three liver fibrosis indices.

Prevalence of steatosis and insulin resistance in patients with chronic hepatitis B compared with chronic hepatitis C and non-alcoholic fatty liver disease

The association of NAFLD with chronic hepatitis C (CHC) has been extensively studied but little is known about its coexistence with chronic hepatitis B (CHB). AimsTo investigate the prevalence and determinants of steatosis and insulin resistance (IR) in CHB and its consequences on liver injury compared with CHC and NAFLD. MethodsPatients with CHB (N=110), CHC (N=111) and NAFLD (N=136) were evaluated by biomarkers of steatosis (SteatoTest>0.38 as a surrogate for steatosis >5%), IR (HOMA-IR>2.7 as a surrogate for IR) and fibrosis (FibroTest>0.48 as a surrogate for significant fibrosis, ≥F2). ResultsHOMA-IR gradually increased in CHB, CHC and NAFLD: 2.3±1.8; 3±2.6 and 3.8±2.7 (p<0.001). The prevalence of steatosis >5% was 21% (CHB), 43% (CHC) and 82% (NAFLD), (p<0.001). The prevalence of fibrosis≥F2 was 10% (CHB), 42% (CHC) and 21% (NAFLD), p<0.001.In CHB, IR was related to host and not viral factors. CHB patients with steatosis had higher BMI (29±5.7kg/m2 vs. 24±4kg/m2, p<0.001), waist circumference (96±14cm vs. 84±11cm, p=0.001) and HOMA-IR (3.9±2.6 vs. 1.8±1.2, p<0.001) than those without steatosis. HOMA-IR independently predicted steatosis in CHB (OR=1.9, 95% CI, 1.09–3.27, p<0.05) and CHC (OR=1.38; 95% CI, 1.07–1.78, p<0.02). In CHB, metabolic risk factors and HOMA-IR were not associated with significant fibrosis. HOMA-IR was an independent predictor of fibrosis in CHC. ConclusionsSteatosis may co-exist in CHB patients but with a lower prevalence than in CHC and NAFLD. In CHB steatosis is related to host and not viral factors, and is not associated with the severity of fibrosis.

AST/ALT ratio is not an index of liver fibrosis in chronic hepatitis C when aminotransferase activities are determinate according to the international recommendations

The aspartate aminotransferase activity (AST)/alanine aminotransferase activity (ALT) ratio is used as liver fibrosis index whereas the reported data are conflicting. In chronic hepatitis C (CHC), reported diagnostic accuracies range from none to good for significant fibrosis and to excellent for cirrhosis. Assuming that AST/ALT increases are mainly due to vitamin B6 defects since pyridoxal phosphate (PLP), active form of B6, acts as coenzyme in transamination reactions, we evaluated the diagnostic accuracy of the AST/ALT ratio using standardized methods for AST and ALT activities, with PLP addition as recommended, in a prospective multicenter cohort of CHC patients. ALT and AST activities were measured using the recommended IFCC methods with addition of pyridoxal 5'-phosphate. We evaluated the AST/ALT ratio for the diagnosis of liver fibrosis or cirrhosis in a cohort of CHC patients included in a multicenter prospective study. A liver biopsy was performed in each patient and reviewed by two independent pathologists in order to determine the fibrosis stage according to Metavir classification which was the reference standard. AST/ALT ratio significantly increased with histological stage of liver fibrosis and there was a significant correlation between Metavir fibrosis stage and AST/ALT ratio (r=0.129, P<0.0035). The ROC curve analyses showed that the AST/ALT ratio does not discriminate significant fibrosis (F≥2) (AUROC=0.531) and had only very poor diagnostic accuracies for severe fibrosis (F≥3) (AUROC=0.584) or cirrhosis (F4) (AUROC=0.626). AST/ALT ratio is not a good and discriminative index of liver fibrosis in CHC when aminotransferase activities are determinate according to the international recommendations.