Non-viral Liver Research Articles

Non-viral liver diseases: pathogenetic approaches to therapy

Alcoholic liver disease and non-alcoholic fatty liver disease are chronic non-communicable diseases with an extremely high prevalence among the world's population. Currently, there is a tendency to a rapid increase in their number with frequent disability of patients and a high need for liver transplantation. Violations of the main metabolic processes in the liver under the influence of exogenous and endogenous factors are the starting point in the development of alcoholic and nonalcoholic liver pathology. This review discusses the main recommendations for non-drug and drug management of patients with liver diseases of non-viral etiology with a detailed description of the main hepatoprotectors with an emphasis on the universal properties of silymarin.

Effect of sample type and storage conditions on Golgi membrane protein 73 stability

GP73 is a transmembrane glycoprotein that increases in viral and non-viral liver diseases, especially in hepatocellular carcinoma. This study aims to evaluate the effect of sample type and storage conditions on GP73 concentration. Twenty subjects were enrolled in this study. Serum and citrated plasma samples were collected. Both were subjected to different time intervals and storage temperature. Baseline GP73 concentrations ranged from 1.7 to 16.9 ng/mL in serum samples, and from 1.1 to 15.3 ng/mL in citrated plasma (Mann–Whitney U test, p = .1). The acceptable change limit for GP73 was 6.1%. As the highest value of the median percentage deviation was −5.3% in both sample types at different storage condition so, deviations were within the accepted limits. But there were considerable variations in the GP-73 concentrations after 2 cycles of freezing and thawing at −20 °C. This study shows that both serum and citrated plasma can be used for the measurement of GP73 concentration. GP73 seems to be stable under common storage conditions, but it may be unstable with frequent cycles of freezing and thawing.

Genetic and lifestyle risk factors for advanced liver disease among men and women.

Liver disease is traditionally categorized as alcoholic and non-alcoholic. We studied various risk factors predictive of advanced non-viral liver disease in general population and analyzed the interaction between these factors and alcohol consumption. Persons without underlying liver disease who participated in the Health2000 or FINRISK studies 1992-2012 comprised a cohort of 41260 individuals. Pattern of alcohol consumption and metabolic, lifestyle-related, and anthropometric parameters were analyzed with Cox regression analysis using severe liver disease hospitalization, cancer, or death as end-point. Viral liver diseases were excluded. A total of 355 liver events occurred during the mean 12.4-year follow-up (511789 person-years). In the multivariate model, age (hazard ratio [HR] 1.03, P=0.0083 for men; HR 1.04, P=0.0198 for women), waist-to-hip ratio (WHR) (HR 1.52, P=0.0006 for men; HR 1.58, P=0.0167 for women), patatin-like phospholipase-containing domain 3 mutations (HR 1.9, P=0.024 for men; HR 2.7, P=0.0109 for women), and weekly binge drinking (HR 2.4, P=0.0024 for men; HR 7.4, P<0.0001 for women) predicted development of severe liver disease. Among men, diabetes (HR 2.7, P=0.0002), average alcohol consumption (HR for 10g/day 1.1, P=0.0022), non-married status (HR 1.9, P=0.0397 for single; HR 2.4, P=0.0002 for widowed/separated), and serum high-density lipoprotein (HR 2.2, P=0.0022) and non-high-density lipoprotein cholesterol (HR 1.2, P=0.0237) were additional risk factors. Alcohol intake increased the risk especially among persons with high WHR (P for interaction 0.009). Age, patatin-like phospholipase-containing domain 3 haplotype, and WHR increase the risk for development of severe liver disease. We found strong synergism between alcohol and central obesity. Binge drinking is an additional risk factor.

Baseline risk factors determine lack of biochemical response after SVR in chronic hepatitis C patients treated with DAAs.

Liver function tests (alanine aminotransferase, ALT; gamma-glutamyltransferase, GGT) not always normalize after elimination of hepatitis C virus (HCV) by direct acting antivirals (DAAs), possibly indicating concomitant non-viral liver diseases. We analysed factors determining the biochemical response (normalized ALT/GGT) of DAA therapy in a large real-world cohort. The German Hepatitis C-Registry is a national multicenter registry study. Normal ALT was defined ≤35U/L (female) and ≤50U/L (male) or, according to AASLD, ≤19U/L (female) and ≤30U/L (male), normal GGT ≤40U/L (female) and ≤60U/L (male). At baseline, ALT was elevated in 3705/4946 (74.9%), ALT (AASLD) in 4669/4946 (94.4%) and GGT in 3018/4906 (61.5%). In this study, 97% of patients achieved SVR12. At week 12 after end of therapy, ALT was elevated in 451/4946 (9.1%), ALT according to AASLD in 1906/4946 (38.5%) and GGT in 863/4879 (17.7%). Persistently elevated ALT after DAA therapy was independently associated with high body mass index (BMI), age <70years, liver cirrhosis, diabetes, alcohol consumption and not achieving SVR12. Using the stricter AASLD criteria, opioid substitution and male sex were additional predictors. Higher GGT at week 12 was associated with high BMI, age >70years, liver cirrhosis, diabetes, alcohol consumption, opioid substitution and non-SVR. Importantly, persistently elevated liver tests after treatment, particularly GGT, were associated with hepatic decompensation and mortality during 4-years follow-up. Risk factors at baseline (obesity, diabetes, liver cirrhosis, alcohol consumption) are independently associated with persistently elevated liver function tests after SVR, indicating that these patients warrant further hepatological follow-up. German Clinical Trials Register (DRKS; ID DRKS00009717).

Combined Effects of Alcohol and Metabolic Disorders in Patients With Chronic Liver Disease

Alcoholic liver disease (ALD) and nonalcoholic fatty liver disease (NAFLD) are the major causes for nonviral liver cirrhosis in the population. Whereas the typical NAFLD patient is one with abdominal obesity, metabolic syndrome (MetS), and no or minimal alcohol use, the patient with pure alcoholic liver cirrhosis has, according to cohort studies, typically consumed >5-10 daily alcohol drinks for several years.1 However, both alcohol use and components of the MetS are continuous variables and, as such, not dichotomic. Recent evidence suggests harmful synergistic effects of obesity, MetS, and alcohol intake for the risk of future liver disease.2 Consequently, given an increasing population prevalence of overweight and obese alcohol users, expectedly, there will be many patients that do not fit either the typical NAFLD or typical ALD phenotype, but share features of both disease entities. Current case-finding strategies focusing on either pure NAFLD or pure ALD3,4 may underestimate the true risk in individuals who will develop liver disease as the result of interaction between alcohol and metabolic disorders.1.

Development and validation of a prediction model to estimate the risk of liver cirrhosis in primary care patients with abnormal liver blood test results: protocol for an electronic health record study in Clinical Practice Research Datalink

BackgroundDriven by alcohol consumption and obesity, the prevalence of non-viral liver disease in the UK is increasing. Due to its silent and slow nature, the progression of liver disease is currently unpredictable and challenging to monitor. The latest National Institute for Health Care Excellence cirrhosis guidelines call for a validated risk tool that would allow general practitioners to identify patients that are at high risk of developing cirrhosis.MethodsUsing linked electronic health records from the Clinical Practice Research Datalink (a database of > 10 million patients in England), we aim to develop and validate a prediction model to estimate 2-, 5- and 10-year risk of cirrhosis. The model will provide individualised cirrhosis risk predictions for adult primary care patients, free from underlying liver disease or viral hepatitis infection, whose liver blood test results come back abnormal. We will externally validate the model in patients from 30 further Clinical Practice Research Datalink general practices in England.DiscussionThe prediction model will provide estimates of cirrhosis risk in primary care patients with abnormal liver blood test results to guide referral to secondary care, to identify patients who are in serious need of preventative health interventions and to help reassure patients at low risk of cirrhosis in the long term.

Interferon Lambda and Liver Fibrosis.

Fibrosis is a highly conserved and coordinated wound healing response to injury. In the liver, injury is promoted by immune effector mechanisms that are common across various disease etiologies and even between organs such as lungs, kidneys, heart, and other organs. Thus, the liver represents a useful model to study inflammation and repair, particularly as it is frequently biopsied in clinical contexts. Currently, strong evidence implicates IFNL3/4 polymorphisms and interferon (IFN)-λ3 levels as determinants of the extent of hepatic inflammation and fibrosis in viral and nonviral liver diseases, as well as in governing the severity of nonhepatotropic viral diseases. Interestingly, IFNL3/4 polymorphisms and IFN-λ3 levels correlate with fibrosis extent in other organs such as the lung and kidney. In this review, we discuss the association between IFN-λ and tissue inflammation and fibrosis in human disease and the potential clinical utility of the findings.

Aspirin: Does it Have a Role for Chemoprevention of Hepatocellular Carcinoma?

Aspirin: Does it Have a Role for Chemoprevention of Hepatocellular Carcinoma?

Multiple primary Malignancies in patients with HCC; Is it a poor prognosis factor for overall survival?

Introduction: Multiple primary malignancies are defined as two or more different malignancies detected synchronously or metachronously in different organs in a single patient. However, studies for the clinicopathologic features regarding multiple primary malignancies in patients with HCC are insufficient. In this study, we performed a retrospective study to investigate the clinicopathologic features for the HCC patients with multiple primary malignancies. Method: Between May 1997 and July 2016, 1043 HCC patients received radical surgical treatment including hepatectomy and liver transplantation in our institute. Among them, 58 (5.6%) cases were diagnosed with extrahepatic primary malignancies. The comparison between single primary HCC and multiple primary malignancies was performed. Result: The five most common extrahepatic primary malignancies are stomach, lung, colorectal, renal and bladder cancers. The patients with multiple primary tumors had a significantly longer overall survival, compared to those with single primary HCCs. The 5-year OS rates for multiple and single primary tumor were 77.8% and 66.9%, respectively. The multiple primary tumor patients had the following clinicopathologic features: older mean age, more patients with non-viral background liver, fewer patients with liver cirrhosis and more patients with AJCC stage I for HCC. Conclusion: The patients with multiple primary tumors have a relatively good prognosis compared to those with no extrahepatic malignancy. This is thought to be mostly due to non-viral background liver and less invasive tumor characteristics.

Outcomes of immunosuppression minimization and withdrawal early after liver transplantation.

The Immune Tolerance Network ITN030ST A-WISH assessed immunosuppression withdrawal in liver transplant recipients with hepatitis C or nonimmune nonviral liver disease. Of 275 recipients enrolled before transplantation, 95 were randomly assigned 4:1 to withdrawal (n=77) or maintenance (n=18) 1- to 2-years posttransplant. Randomization eligibility criteria included stable immunosuppression monotherapy; adequate liver and kidney function; ≤Stage 2 Ishak fibrosis; and absence of rejection on biopsy. Immunosuppression withdrawal followed an 8-step reduction algorithm with ≥8weeks per level. Fifty-two of 77 subjects (67.5%) reduced to ≤50% of baseline dose, and 10 of 77 (13.0%) discontinued all immunosuppression for ≥1year. Acute rejection and/or abnormal liver tests were treated with increased immunosuppression; 5 of 32 rejection episodes required a methylprednisolone bolus. The composite end point (death or graft loss; grade 4 secondary malignancy or opportunistic infection; Ishak stage ≥3; or >25% decrease in glomerular filtration rate within 24months of randomization) occurred in 12 of 66 (18%) and 4 of 13 (31%) subjects in the withdrawal and maintenance groups. Early immunosuppression minimization is feasible in selected liver recipients, while complete withdrawal is successful in only a small proportion. The composite end point comparison was inconclusive for noninferiority of the withdrawal to the maintenance group.

Relationship between genetic variation at PPP1R3B and levels of liver glycogen and triglyceride.

These observations are consistent with the notion that the minor allele of rs4841132 promotes a mild form of hepatic glycogenosis that is associated with hepatic injury. (Hepatology 2018;67:2182-2195).

Soluble intercellular adhesion molecule-1 is associated with hepatocellular carcinoma risk: multiplex analysis of serum markers

Individualized assessment of hepatocellular carcinoma (HCC) risk in chronic liver disease remains challenging. Serum biomarkers including cytokines may offer helpful adjuncts to standard parameters for risk prediction. Our aim was to identify markers associated with increased HCC incidence. This was a prospective cohort study of 282 patients with both viral or non-viral chronic liver disease. Baseline serum cytokines and other markers were measured in multiplex with a commercially-available Luminex-based system. Patients were followed until death or HCC diagnosis. We performed Lasso-based survival analysis to determine parameters associated with HCC development. Cytokine mean florescence intensity (MFI) was the primary predictor and HCC development the primary outcome. 25 patients developed HCC with total follow-up of 1,363 person-years. Parameters associated with increased HCC incidence were cirrhosis, hepatic decompensation, and soluble serum intercellular adhesion molecule 1 (sICAM-1) MFI. No other molecules increased predictive power for HCC incidence. On univariate analysis, the parameters associated with HCC incidence in patients with cirrhosis were age, antiviral treatment, and high sICAM-1 MFI; on multivariate analysis, sICAM-1 remained associated with HCC development (adjusted HR = 2.75). On unbiased screening of serum cytokines and other markers in a diverse cohort, baseline sICAM-1 MFI is associated with HCC incidence.

Proteoforms in Peripheral Blood Mononuclear Cells as Novel Rejection Biomarkers in Liver Transplant Recipients.

Biomarker profiles of acute rejection in liver transplant recipients could enhance the diagnosis and management of recipients. Our aim was to identify diagnostic proteoform signatures of acute rejection in circulating immune cells, using an emergent "top-down" proteomics methodology. We prepared differentially processed and cryopreserved cell lysates from 26 nonviral liver transplant recipients by molecular weight-based fractionation and analyzed them by mass spectrometry of whole proteins in three steps: (i) Nanocapillary liquid chromatography coupled with high-resolution tandem mass spectrometry; (ii) database searching to identify and characterize intact proteoforms; (iii) data processing through a hierarchical linear model matching the study design to quantify proteoform fold changes in patients with rejection versus normal liver function versus acute dysfunction without rejection. Differentially expressed proteoforms were seen in patients with rejection versus normal and nonspecific controls, most evidently in the cell preparations stored in traditional serum-rich media. Mapping analysis of these proteins back to genes through gene ontology and pathway analysis tools revealed multiple signaling pathways, including inflammation mediated by cytokines and chemokines. Larger studies are needed to validate these novel rejection signatures and test their predictive value for use in clinical management.

Zinc is a potent and specific inhibitor of IFN-\u03bb3 signalling

Lambda interferons (IFNL, IFN-λ) are pro-inflammatory cytokines important in acute and chronic viral infection. Single-nucleotide polymorphisms rs12979860 and rs8099917 within the IFNL gene locus predict hepatitis C virus (HCV) clearance, as well as inflammation and fibrosis progression in viral and non-viral liver disease. The underlying mechanism, however, is not defined. Here we show that the rs12979860 CC genotype correlates with increased hepatic metallothionein expression through increased systemic zinc levels. Zinc interferes with IFN-λ3 binding to IFNL receptor 1 (IFNLR1), resulting in decreased antiviral activity and increased viral replication (HCV, influenza) in vitro. HCV patients with high zinc levels have low hepatocyte antiviral and inflammatory gene expression and high viral loads, confirming the inhibitory role of zinc in vivo. We provide the first evidence that zinc can act as a potent and specific inhibitor of IFN-λ3 signalling and highlight its potential as a target of therapeutic intervention for IFN-λ3-mediated chronic disease.

Research advances in autoimmune liver diseases in 2016

Autoimmune liver diseases are a group of abnormal autoimmune-mediated inflammatory hepatobiliary injuries, mainly including autoimmune hepatitis(AIH), primary biliary cholangitis(PBC), and primary sclerosing cholangitis (PSC). The diagnosis and treatment of autoimmune liver diseases, an important type of non-viral liver disease, have become a prominent issue in hepatology. In 2016, many new advances have been achieved in the clinical and basic research on autoimmune liver diseases, including the phase 3 clinical trial of obeticholic acid, the proposal of UK-PBC risk score, and the research on gut microbiota associated with PSC. This article reviews the research advances in the diagnosis and treatment of autoimmune liver diseases in 2016.

Patients with non-viral liver disease have a greater tumor burden and less curative treatment options when diagnosed with hepatocellular carcinoma.

AIMTo assess the impact of underlying liver disease etiology on the presenting features and outcomes in a large cohort of patients with hepatocellular carcinoma (HCC).METHODSA prospective database of all patients with HCC was established from 1998 to March 2012. One thousand and seventy-eight patients were categorized into three groups, based on the etiology of their liver disease: hepatitis B virus (HBV), hepatitis C virus (HCV) and non-viral liver disease (NVLD). Overall survival was determined by Kaplan Meier analysis to time of death or last follow-up.RESULTSHCC patients with HCV (85%) were more likely to be diagnosed as part of a surveillance program, compared to HBV or NVLD (both 71%) (P < 0.001). Patients with NVLD were more likely to receive best supportive care (29%) compared to those with HBV (21%) or HCV (20%) (P < 0.02). Twelve percent of NVLD and 13% of HBV patients underwent liver transplantation compared to 21% of HCV patients (P = 0.001). Median survival from presentation was lowest in NVLD (1.7 years) when compared to HBV (2.8 years) and HCV (2.6 years) (P < 0.05). In multivariate analysis, independent predictors of survival included Child Turcotte Pugh score, size of dominant lesion, absence of vascular invasion, and management with surgical resection or liver transplantation. Patient age and the etiology of the underlying liver disease were not independent predictors of survivalCONCLUSIONPatients with NVLD and HCC were less likely to be enrolled in a HCC surveillance program and are less likely to have curative therapies such as liver resection and transplantation after diagnosis with HCC, when compared to patients with Hepatitis B and Hepatitis C.

Association of Serum Ferritin with Diabetes and Alcohol in Patients with Non-Viral Liver Disease-Related Hepatocellular Carcinoma

Introduction: Non-alcoholic fatty liver disease is a leading cause for hepatocellular carcinoma (HCC) in Sri Lanka. Diabetes mellitus, alcohol abuse, and liver inflammation are known to increase the risk of HCC. The present study evaluates serum ferritin levels in a cohort of patients with non-viral HCC (nvHCC). Methodology: Consecutive patients with nvHCC presenting to the Colombo North Liver transplant Service, Ragama, from January 2012 to July 2013 were investigated. All were negative for hepatitis B and C. At registration, 5 mL of serum was separated into plain tubes, stored at -80°C and analysed for ferritin using an enzyme-linked immunosorbent assay. Correlation between the serum ferritin and patient risk factors, liver status, and tumour characteristics were analysed. Results: There were 93 patients with nvHCC (median age 65 [12-82] years; 82 [88.2%] males). The median ferritin level was 246.2 μg/L, and 38 (40.86%) patients had elevated ferritin. Non-diabetics (median 363.5 mg/L, p = 0.003) and alcohol abusers (median 261.2 mg/L, p = 0.018) had higher ferritin levels. On multiple-variable analysis, being non-diabetic (p = 0.013) and alcoholic (p = 0.046) was significantly associated with high serum ferritin. No association was found with body mass index, tumour stage, size, macrovascular invasion, number of nodules, alpha-fetoprotein, bilirubin, international normalized ratio, and survival. Conclusion: In patients with nvHCC, serum ferritin levels are higher in non-diabetics and alcoholics.

Clinical observation on the treatment of acute liver failure by combined non-biological artificial liver.

The clinical efficacy and safety of different combinations of non-bio artificial liver in the treatment of acute liver failure was examined. A total of 61 cases were selected under blood purification treatment from the patients with severe acute liver failure admitted to the severe disease department of the hospital from December, 2010 to December, 2015. Three types of artificial liver combinations were observed, i.e., plasma exchange plus hemoperfusion plus continuous venovenous hemodiafiltration (PE+HP+CVVHDF), PE+CVVHDF and HP+CVVHDF. The heart rate (HR), mean arterial pressure (MAP), respiratory index (PaO2/FiO2), liver and kidney function indicator, as well as platelet and coagulation function were compared. A comparison before and after the treatment using the three methods, showed improvement in the HRs, MAPs, PaO2/FiO2, total bilirubins (TBIL) and alanine aminotransferases (ALT) (P<0.05), of which TBIL and ALT were decreased more significantly (P<0.01) in the PE+CVVHDF and PE+HP+CVVHDF groups. Only changes in the PE+HP+CVVHDF and PE+CVVHDF groups were statistically significant after prothrombin time and albumin treatment (P<0.05). The difference between the decrease in TBIL in the PE+HP+CVVHDF group and that in the HP+CVVHDF group was statistically significant (P<0.05). Treatment of the 61 patients using the artificial liver support system yielded a survival rate of 62.3% (38/61), and a viral survival rate of 35.0% (7/20); with the non-viral survival rate being 75.6% (31/41). In conclusion, following the treatment of three types of artificial livers, the function was improved to varying degrees, with the PE+HP+CVVHDF and the PE+CVVHDF method being better. By contrast, after the treatment of non-viral liver failure, the survival rate was significantly higher than the patients with viral liver failure.

Serum YKL-40 as a marker of liver fibrosis in patients with non-alcoholic fatty liver disease.

Non-alcoholic fatty liver disease (NAFLD) is a common cause of chronic non-viral liver disease. YKL-40, chitinase-like protein expressed in multiple tissues including liver, is involved in cell proliferation, inflammation and remodeling of the extracellular matrix. The aim of this study was to assess whether serum YKL-40 levels are associated with liver fibrosis in NAFLD patients. Serum YKL-40 levels were quantified in 111 NAFLD patients and 23 HCC patients with NAFLD. To identify the source of YKL-40, immunofluorescence staining of liver specimens from NAFLD patients was performed. Serum YKL-40 levels in NAFLD patients increased in accordance with the progression of liver fibrosis. Multivariate analysis revealed that YKL-40 was one of the independent factors significantly associated with severe fibrosis (F3-4). We established a new predictive model for fibrosis of NAFLD, using logistic regression analysis: YKL-40 based fibrosis score = −0.0545 + type IV collagen 7s * 0.3456 + YKL-40 * 0.0024. Serum YKL-40 levels of HCC patients with non-cirrhotic NAFLD were significantly higher than those without HCC. Immunofluorescence staining showed that YKL-40 was expressed by macrophages in liver tissue of NAFLD patients. In conclusion, macrophage-derived YKL-40 is a feasible biomarker of liver fibrosis in NAFLD patients.

Genotyping of coding single nucleotide variants of the hOAT2[SLC22A7] gene in Japanese patients with non-viral liver tumor

Human organic anion transporter 2 (hOAT2[SLC22A7]) is a Na+-independent multispecific organic solutes (SLC) transporter. To date, several findings regarding the function and substrates of hOAT2[SLC22A7] have been reported; there are still limited data concerning the genetic variants of the hOAT2[SLC22A7] gene in Japanese, specifically, in patients with non-viral liver carcinoma (LC). In the present study, we isolated genomic DNA from a normal portion of liver cancer and screened comprehensively 88 coding single nucleotide polymorphisms (cSNPs) chosen from a database of SNPs (dbSNPs). We found genotype and allele frequencies for 1 synonymous SNP [rs2270860 (1324C>T, Ser425Ser)] to be statistically significant when compared with the SNP database (http://www.ncbi.nlm.nih.gov/snp/). Our present findings suggest that, at least partly, within the entire sequence of coding region of the hOAT2[SLC22A7] gene, rs2270860 (1324C>T, Ser425Ser) might be useful as a biomarker for susceptibility for non-viral LC in Japanese.