Search

Background/Aims Response to HCV treatment with pegylated interferon-α is variable but might at least in part depend on genetic host factors. The G protein β3 unit (GNB3) C825T polymorphism has been shown to affect treatment response in HCV mono-infection. Here, we analyzed the impact of the GNB3 genotype in the context of HCV/HIV co-infection. Methods HIV/HCV co-infected (n = 112) and HCV mono-infected patients (n = 150), receiving therapy with pegylated IFN-α/ribavirin, were enrolled into this study. Furthermore, we analyzed 220 healthy and 92 HIV mono-infected patients. GNB3 genotype was defined and correlated with respect to treatment response. Results GNB3 genotype distribution differed significantly between HIV/HCV co-infected patients and HIV-positive/HCV-negative (p = 0.0002) or healthy controls (p = 0.03). Patients with a GNB3 CC genotype had significantly lower SVR rates as compared to carriers of a non-CC genotype (52% versus 77%; p = 0.018). In a logistic regression analysis the GNB3 genotype and the HCV genotype were significantly associated with response to treatment (p = 0.018). In contrast to HIV/HCV co-infected patients, GNB3 genotype did not affect response to treatment in HCV mono-infected patients. Conclusions The GNB3 825 CC genotype is associated with poor SVR rates in HIV/HCV co-infected patients. This underlines the impact of genetic host factors for treatment response.

Background/Aims : The CC-chemokine receptor (CCR) 5-Δ32 mutation may predispose to chronic liver disease and high level viremia in hepatitis C. However, it is unclear whether CCR5-Δ32 also affects the response to antiviral treatment. Methods : We determined CCR5 genotypes in patients with hepatitis C treated with either interferon-α ( N =78) or interferon and ribavirin ( N =78). In each group, rates of end of treatment responses (ETRs) and sustained virological responses (SVRs) were compared between CCR5-Δ32 carriers and homozygous CCR5 wildtype patients. Results : ETR and SVR were achieved in 25 and 12 patients with interferon-α and in 52 and 45 patients with interferon/ribavirin treatment, respectively. CCR5-Δ32 carriers had significantly lower ETR rates than homozygous CCR5 wildtype patients (10.5 vs. 39.0%; P =0.02), whereas SVR rates only showed a non-significant trend (5.3 vs. 18.6%). Multivariate analysis confirmed CCR5-Δ32 carriage as an independent negative predictor for ETR in interferon-α monotherapy (odds ratio: 0.16; 95% confidence limits: 0.032–0.82; P =0.03). In interferon/ribavirin treated patients CCR-Δ32 carriers and CCR5 wildtype patients had similar ETR rates [19.2% vs. 23.1%] and SVR rates [20.0% vs. 21.2%]. Conclusions : Response rates to interferon-α monotherapy are reduced in hepatitis C virus (HCV)-infected patients carrying the CCR5-Δ32 mutation. However, interferon/ribavirin combination treatment may overcome this negative effect of CCR5-Δ32.

Background/Aims: Common genetic variations in vitamin D metabolism are associated with liver stiffness. Whether these genes are implicated in hepatic steatosis remains unclear. Here we aimed to analyse the association of common vitamin D pathway gene variants with liver steatosis. Methods: Liver steatosis was assessed non-invasively in 241 patients with chronic liver conditions by controlled attenuation parameter (CAP). The following polymorphisms were genotyped using TaqMan assays: group-specific component (GC) rs7041, 7-dehydrocholesterol reductase (DHCR7) rs12785878, cytochrome P450 2R1 (CYP2R1) rs10741657, ­vitamin D receptor (VDR) rs7974353. Chemiluminescence immunoassay determined serum 25-hydroxyvitamin D (25(OH) D) concentrations. Results: Vitamin D deficiency (defined by 25(OH)D concentrations p = 0.033) lower 25(OH)D levels as compared to patients with CAP GC rs7041 was significantly (p = 0.018) associated with higher 25(OH)D levels in patients with CAP GC, DHCR7, CYP2R1, and VDR polymorphisms were not related to liver steatosis and obesity traits. Conclusions: Higher CAP values are associated with low serum 25(OH)D concentrations but not with common vitamin D pathway gene variants.

Background & Aims CXCL1 is a ligand for CXC chemokine-receptor 2 expressed on hepatic stellate cells (HSC). Thus, CXCL1 might contribute to HSC activation and fibrogenesis. Here, we investigated whether the CXCL1 rs4074 polymorphism affects CXCL1 expression and progression of chronic hepatitis C virus (HCV) infection towards cirrhosis. Methods The study involved 237 patients with chronic HCV genotype 1 infection (75 with cirrhosis) and 342 healthy controls. The CXCL1 rs4074 polymorphism was determined by a LightSNiP assay on the LightCycler system. CXCL1 serum levels and induction in response to HCV proteins were studied by ELISA. Results Distributions of CXCL1 genotypes (GG/GA/AA) matched the Hardy–Weinberg equilibrium in all subgroups (HCV-associated cirrhosis: 29.3%/54.7%/16.0%; non-cirrhotic HCV infection: 45.1%/44.4%/10.5%, healthy controls: 46.2%/40.9%/12.9%). HCV-infected cirrhotic patients had a significantly greater CXCL1 rs4074 A allele frequency (43.3%) than patients without cirrhosis (32.7%, OR=1.573, p =0.03) and healthy controls (33.3%, OR=1.529, p =0.02). In vitro carriers of the A allele produced greater amounts of CXCL1 in response to TLR2-ligands including HCV core and NS3, and HCV-infected carriers of the CXCL1 rs4074 A allele had higher CXCL1 serum levels than those with the G/G genotype. Moreover, multivariate Cox-regression analysis confirmed age and the presence of a CXCL1 rs4074 A allele as risk factors for cirrhosis. Conclusions Enhanced production of CXCL1 in response to HCV antigens in carriers of the rs4074 A allele together with its increased frequency in cirrhotic patients with hepatitis C suggest the CXCL1 rs4074 A allele as a genetic risk factor for cirrhosis progression in hepatitis C.

Background & Aims Toll-like receptor (TLR) 2 and nucleotide-binding oligomerisation domain (NOD) 2 recognize distinct pathogen-associated molecular patterns (PAMS) on the cell surface and in the cytoplasm, respectively. Since they may contribute to susceptibility to spontaneous bacterial peritonitis (SBP), we studied the effects of TLR2 gene variants on susceptibility for SBP in relation to the previously reported NOD2 alleles. Methods Overall, 150 patients with liver cirrhosis and ascites were genotyped for TLR2 gene variants −16934 (rs4696480), Arg753Gln (rs5743708), Pro631His (rs5743704) and the TLR2 GT microsatellite polymorphism in intron 2. Patients were monitored for SBP over two years. TLR2 SNPs were identified by hybridization probe assays on a LightCycler system. Numbers of GT repeats were determined with an ABI310 sequencer and Genescan Analysis 2.1 software. Results Fifty two patients (35%) had SBP. Unlike the TLR2 Arg753Gln and Pro631His mutations, SBP was significantly more frequent in patients with the TLR2 −16934 TT genotype (38.5% vs. 15.3%; p =0.002) and in carriers with two long tandem GT repeat alleles (>20) (53.8% vs. 25.5%; p =0.001). A multivariate analysis confirmed TLR2 GT microsatellite polymorphism (OR=3.8, p =0.002) and NOD2 variants (OR=3.3, p =0.011) as independent predictors of SBP, and the simultaneous presence of both risk factors indicated a particularly high risk for SBP (OR=11.3, p =0.00002). Conclusions Analogous to NOD2 risk variants, TLR2 polymorphisms indicate increased susceptibility toward SBP in cirrhotic patients with ascites, and the combination of the TLR2 GT microsatellite polymorphism with at least one NOD2 risk variant enables improved identification of patients with a high risk for SBP.

Background & Aims Recent genome-wide association studies have identified the variant p.I148M of the adiponutrin gene PNPLA3 as a risk factor for developing severe forms of non-alcoholic and alcoholic liver diseases. The risk allele confers an increased risk for fatty liver disease and elevated serum aminotransferase activities reflecting liver injury. In the current elastography-based study, we investigate variant adiponutrin as genetic determinant of liver fibrosis, the hallmark of all chronic liver diseases. Methods In this observational cross-sectional study, we staged 899 patients with different chronic liver diseases non-invasively by transient elastography (Fibroscan) and genotyped them for variant adiponutrin (rs738409) by PCR-based assays. A subgroup of 229 patients consented to percutaneous liver biopsy, validating the accuracy of elastography in staging fibrosis ( ρ =0.743, p Results Carriers of distinct p.I148M adiponutrin genotypes display significant ( p =0.017) differences in liver stiffness determined by elastography. In particular, individuals carrying the allele [G] are at higher risk of developing liver cirrhosis defined by stiffness values ⩾13.0kPa (OR=1.56, p =0.005). Of note, the PNPLA3 risk variant advances fibrosis in the total cohort as well as in the subgroups of patients with viral hepatitis and non-viral liver diseases and contributes 16% of the total cirrhosis risk. Conclusions The adiponutrin risk variant is a common genetic determinant of progressive liver fibrosis. Our results underpin non-invasive follow-up for individuals with chronic liver disease at-risk for developing advanced fibrosis and cirrhosis.

Recently, variation of gene copy numbers was recognized as a novel type of common genetic diversity, but its impact on viral hepatitis is unknown. Here, we determine the influence of copy number variation on the susceptibility and disease severity in hepatitis C virus (HCV) infection, investigating copy number variants (CNVs) of the chemokine CCL3L1 gene, which encodes a potent CCR5 ligand.CNVs were determined in 254 patients with chronic hepatitis C, 144 HCV/HIV co-infected patients, and 210 HCV negative controls, using quality-controlled real-time fluorescent dye-labeled quantitative PCR. Liver biopsies were obtained from HCV infected patients.Copy numbers of the CCL3L1 gene range from 0 to 12 (mean 2.7+/-1.4 copies). Patients with two or less copies are over-represented in the HCV infected cohort compared to HCV negative controls (odds ratio [OR] 1.54; p=0.02). CCL3L1 copies are shifted to lower numbers in HCV infected patients (means 2.6 vs. 2.9 in controls; p=0.011). HCV/HIV co-infected patients carry even lower CCL3L1 copy numbers compared to controls (means 2.2 vs. 2.9; p0.001), with a higher proportion of patients possessing two or less copies (OR=3.42; p0.001). No association was detected between CCL3L1 copy numbers and histological grades of inflammation or stages of fibrosis.Lower CCL3L1 gene copy number compared to the population median is associated with chronic hepatitis C. Copy number variation of host genes represents a novel class of genetic diversity associated with viral hepatitis.

(Mdr2-KO, bile duct ligation, treatment with dehydrocollidine – DDC). However, PC-2-gene expression was not reduced, suggesting that the decrease in PC2 protein content was due to increased degradation. To understand if factors involved in biliary damage influence PC2 protein expression, mouse cholangiocytes were treated with pro-inflammatory cytokines (IL-1b, IFNg, TNFa), nitric oxide (NO) donors (DETAnonoate) and ER stressors (thapsigargin). Expression of PC2 protein was again significantly reduced, but not its gene expression. Noteworthy, downregulation of PC-2 was associated to increased ERK1/2 phosphorylation, HIF1a transcription activity, secretion of VEGF and phosphorylation of VEGFR2. Expression of Herp and NEK, ubiquitin-like proteins that promote PC2 degradation by increasing the retrotranslocation from the ER to the proteasome complex was also increased. Pre-treatment with the proteasome inhibitor MG-132 restored the expression of PC2 in cells treated with cytokines but not in cells treated with NO donors or with ER stressors. In the latter conditions, PC2 degradation was inhibited by LY294002, an inhibitor of PI3K that blocks the autophagosome formation and by blockade of lysosomes with chloroquine, suggesting a role for autophagy. Finally, treatment of DDC-treated mice with the proteasome inhibitor bortezomib, significantly reduced the extent of the liver damage and restored PC2 expression. Conclusions: PC2 protein is modulated post-translationally by proinflammatory cytokines, ER-stressors and NO-donors and is reduced in mice with biliary damage. Down regulation of PC2 protein expression in cholangiocytes increases the ERK1/2/HIF1a pathway and VEGF secretion, thereby playing a pivotal role in the regulation of cholangiocyte response to biliary damage. Treatments able to restore PC2 expression (i.e. proteasome inhibitors) may represent a new therapeutic approach in biliary diseases.

690 PATATIN-LIKE PHOSPHOLIPASE-3 (PNPLA3) VARIATION AND BILIARY TRACT CANCER RISK V. Zimmer, A. Hoblinger, F. Mihalache, M. Krawczyk, F. Grunhage, M. Acalovschi, F. Lammert. Department of Medicine II, Saarland University Medical Center, Homburg, Department of Medicine I, University of Bonn, Bonn, Germany; Department of Medicine III, University Iuliu Hatieganu, Cluj-Napoca, Romania E-mail: vincent.zimmer@uks.eu

Figure 2. (a) Ex-vivo monocyte subsets differentiation at 0 hour, 24 hours, 36 hours in a survival ALF patient on the 1, 3, 7 day after admission; The Mo1/Mo2 ratio was 4.78 (day 1), 1.54 (day 3), 0.55 (day 7), respectively, which suggested that the repressed monocyte subsets differentiation on the 1 and the 3 day, then, recovered on 7 day in the survivor. (b) Main clinical data for the patient on the 1, 3, 7 day after admission.