- Email: [email protected]
Patients with Chronic Hepatitis C Can be Identified by their Gut Microbiota
POSTER PRESENTATIONS complications. The reasons why hepatitis C promotes insulin resistance are partially known. Aims: To evaluate a wide multitest panel including biochemical, metabolic, inflammatory, genetic and virologic parameters in patients with hepatitis C and to identify factors related with insulin resitance.
Background and Aims: The importance of the intestinal microbiota for the onset and clinical course of many diseases, including liver diseases like NASH and liver cirrhosis, is increasingly recognized. However, the role of intestinal microbiota in chronic HCV infection remains unclear. Methods: In a cross sectional approach the intestinal microbiota of 95 patients chronically infected with HCV (n = 57 w/o cirrhosis (NOCIR); n = 38 with cirrhosis (CIR) and 50 healthy controls (HC) was analysed. The V1-2 region of the 16S rRNA gene was amplified followed by sequencing on the Illumina MiSeq platform. Random subsampling was performed to achieve equal number of reads.
Methods: Patients with chronic hepatitis C without any antiviral therapy in the previous one year were included. Subjects with decompensated cirrhosis, hepatitis B o HIV coinfection, or previous diagnosis of diabetes mellitus were excluded. In 76 patients, we performed a blood laboratory test which include routine, metabolic and inflammatory parameters (retinol, retinol-binding protein 4, 25OH vitamin D, Vitamin E, lipopolysaccharide-binding protein, interleukin-6 and cystatin C). Insulin resistance was established with a HOMA value higher than 3. We determined single nucleotide polymorphisms rs7041 and rs4588 GC-DBP (Group specific component-Vitamin D-binding protein), rs 738409 PNPLA3 ( patatin-like phospholipase domain containing 3), and rs12979860 IL28B (interleukin-28 B) genes. Viral parameters like genotype o viral load were obtained and fibrosis staging was assessed with liver biopsy or transient elastography. Results: After backward logistic regression analysis, independent variables associated with insulin resistance were: 1) Gc1s/Gc1S Vitamin D-binding protein phenotype, that results from the homozygous carriage of the rs7041G/rs4588C haplotype ( p = 0.033); 2) low retinol/RBP4 ratio, reflecting a greater rate of unbound retinol-binding protein 4 ( p = 0.005); 3) older age ( p = 0.01); 4) high serum tryglicerides ( p = 0.026), and 5) advanced (F3F4) fibrosis stage ( p = 0.034). The AUROC provided by the multivariate model was 0.950 (95 % IC = 0.906–0.993). Conclusions: In addition to previously known ones, the Gc1s/Gc1s phenotype variant of vitamin D-binding protein and the unbound fraction of plasma retinol-binding protein 4 may be considered as factors related with the incidence, and possibly the risk, of insulin resistance in chronic hepatitis C patients. FRI-159 PATIENTS WITH CHRONIC HEPATITIS C CAN BE IDENTIFIED BY THEIR GUT MICROBIOTA B. Heidrich1,2,3, M. Vital2, I. Plumeier2, S. Kahl2, J. Kirschner1, L. Sollik1, S. Ziegert1, M. Manns1,3, H. Wedemeyer1,3, D. Pieper2,3. 1Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Hannover; 2Microbial Interactions and Processes Research Group, Center for Infection Research, Braunschweig; 3German Center for Infection Research (DZIF), Hannover-Braunschweig, Germany E-mail: [email protected]
Results: The number of phylotypes (S) was significantly lower in NOCIR vs. HC (386 ± 77 vs. 420 ± 72; p = 0.003) and CIR vs. HC (329 ± 86 vs. 386 ± 77; p = 0.009). Shannon diversity index H’ was different between CIR vs. HC (4.0 ± 0.5 vs. 4.4 ± 0.3; p < 0.001) and CIR vs. NOCIR (4.0 ± 0.5 vs. 4.4 ± 0.4; p = 0.002) while no differences were
Journal of Hepatology 2016 vol. 64 | S425–S630
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POSTER PRESENTATIONS observed between HC vs NO-CIR. S and H’ correlated negatively with ARFI (S: R = −0.590; p < 0.001; H’: R = −0.528; p = 0.002) in patients. The presence or absence of 14 genera, which were more often observed in either HC or NO-CIR was used to develop a score to differentiate between HC and NO-CIR. If a genus, that was more often observed in HC, was observed in a sample, it was scored + 1 whereas it was scored −1, when the genus was more often observed in NO-CIR. The total score ranged from −6 to +8. Assuming samples of a score >2 as being from HC, the score has an AUC of 0.917 with a sensitivity of 0.82, specificity of 0.92, PPV of 0.92 and NPV of 0.82. Additionally, we observed four distinct patterns in the relative abundance of genera. The relative abundance of 5 genera increased from HC to NO-CIR to CIR ( pattern 1), whereas the relative abundance of 8 genera decreased, and the abundance of those genera can, thus, be associated with fibrosis progression. Six genera showed a significant difference in abundance in CIR but comparable abundancies in HC and NO-CIR ( pattern 3), and their abundance could, thus be associated with cirrhosis. A further 6 genera showed a significant difference between HC and NO-CIR ( pattern 4), thus, being associated with HCV infection (Fig). Conclusions: These results demonstrate an association of chronic HCV infection with the community structure in the gut compared to HC even in the absence of cirrhosis. It has to be clarified if these differences in the microbiota are reversible with the clearance of HCV by novel DAAs. FRI-160 RISKS OF LIVER COMPLICATIONS AFTER A LONG-TERM FOLLOW-UP IN PATIENTS WITH CHRONIC HEPATITIS C WITH SUSTAINED VIROLOGIC RESPONSE B. Kutala1, M.-P. Ripault1, E. Estrabaud1, F. Khelifa-Mouri1, R. Nait1, N. Boyer1, N. Giuily1, C. Castelnau1, T. Asselah1, P. Marcellin1. 1Service d’hépatologie, Hôpital Beaujon, Clichy, France E-mail: [email protected] Background and Aims: The long-term effect of sustained virologic response (SVR) to antiviral therapy on the risk of developing hepatocellular carcinoma (HCC), liver decompensation, liver-related death, and overall death, in patients with chronic hepatitis C (HCV), has been determined by few studies in the short term and never in the long term. Methods: The study was conducted on 1024 patients with chronic hepatitis C treated with antiviral regimens based interferon (IFN) who had achieved SVR at the university center of Beaujon from 2000 to 2011. Complementary follow-up data were obtained by contacting patients themselves, their primary care physician, their municipality of birth and the French biomedicine agency for patients on list of liver transplantation. Results: The baseline median (IQR) age was 51 (40–59) years and 734 patients (72%) were men. The advanced fibrosis score (Metavir F3–F4) was in 403 patients (39%), genotype 1 found in 542 patients (53%). During the follow-up (mean of 5.6 years), among patients with advanced fibrosis (F3–F4), 19 patients died or underwent liver transplantation (LT), 31 patients developed HCC 14 patients had liver decompensation. However in patients without advanced fibrosis (F0–F2) 3, 2 and 2 patients respectively died or underwent LT, developed HCC and had liver decompensation. The incidences of overall death or liver transplantation, HCC and any liver decompensation per 100 person-years were significantly higher in F3-F4 patient time with 0.8, 1.3 and 0.6, compared to 0.2, 0.1, and 0.1 in patients without advanced fibrosis (F0-F2). Conclusions: Among patients with SVR, the risk of death or LT, HCC and liver decompensation were markedly reduced after 3 years follow-up the SVR. However, a long-term risk of developing HCC persists in patients with advanced fibrosis. There was no recurrence of HCV infection or evidence of liver disease progression in patients without advanced fibrosis (F0–F2). Future studies focusing on the
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better identification of patients with a long-term risk for HCC are needed. FRI-161 LONG-TERM ASSESSMENT OF RENAL FUNCTION IN PATIENTS FOLLOWING ANTIVIRAL THERAPY FOR CHRONIC HEPATITIS C VIRUS INFECTION C.-H. Liu1, C.-L. Chen2, C.-J. Liu2, T.-H. Su1, H.-C. Yang1, P.-J. Chen2, D.-S. Chen3, J.-H. Kao3. 1Internal Medicine; 2Graduate Institute of Clinical Medicine; 3Hepatitis Research Center, National Taiwan University Hospital, Taipei, Taiwan E-mail: [email protected] Background and Aims: The long-term progression of renal function in patients following antiviral therapy for chronic hepatitis C virus (HCV) infection remains unclear. Methods: A total of 1,569 HCV-infected patients receiving peginterferon/ribavirin or direct acting antiviral agents (DAAs) were recruited in this prospective cohort study. Serial renal function assessment was evaluated by modification of diet in renal disease equation (MDRD). Linear mixed-effect models were used to estimate the effect of sustained virologic response (SVR) on the changes of MDRD during follow up. Adjusted variables included age, sex, history of DM, hypertension and hyperlipidemia and the interaction between time and SVR. Results: The median time (interquartile range) of post-treatment follow-up was 48 (6–66) months and the SVR rate was 80.1%. The analysis was restricted to 48 months of follow-up. The renal function was similar at the beginning of follow up, regardless of SVR status (82.7 ± 21.4 vs. 80.1 ± 21.8 for SVR vs. non SVR), but significantly lower among patients with DM and hypertension. As time progress, the MDRD started to increase with an average of 0.21 (95% CI = 0.17–0.26) per 6 month. The slope of the increase was higher for those not achieving SVR (averaging 0.25 increase per 6 month, adjusted p for trend <0.001) than those achieving SVR (averaging 0.07 per 6 month, adjusted p for trend = 0.054). Conclusions: In patients undergoing antiviral therapy for HCV, those who achieved SVR was associated with slower progressive loss of renal function. FRI-162 EFFECT OF AMIODARONE AND HCV DIRECT-ACTING ANTIVIRAL AGENTS ON CARDIAC CONDUCTION IN NONCLINICAL STUDIES G. Liu1, C.H. Tay1, S. Rajamani1, A. Ray1, L. Stamm1, J. Vick2, M. Wilichinsky1, J. McHutchison1, D. Brainard1. 1Gilead Sciences, Inc., Foster City; 2ChanTest, a Charles River Company, Ohio, United States E-mail: [email protected] Background and Aims: In the post-market setting, serious cases of symptomatic bradycardia have been reported in patients taking amiodarone with Sovaldi in combination with an HCV direct-acting antiviral (DAA). The underlying mechanism for these events is unknown but is currently under investigation. Amiodarone, a multiion channel inhibitor including Ca2+ currents, has been shown to affect the atrial-to-ventricular conduction precipitating bradycardia. Sofosbuvir, a HCV NS5B polymerase nucleotide inhibitor, did not alter the heart rate, blood pressure, or EKG in telemeterized dogs and up to supratherapeutic doses in a clinical TQT study. The effect of amiodarone and DAAs, alone and in combination, on cardiac conduction in guinea pig isolated hearts, and in vitro inhibition of human calcium channels were determined. Methods: Using guinea-pig isolated hearts, the effect of amiodarone, daclatasvir (a NS5A inhibitor), simeprevir (a NS3/4A protease inhibitor), sofosbuvir and GS-331007 (nucleoside analogue of sofosbuvir) alone or in combination at their maximal concentrations achieved in plasma, on cardiac conduction was determined; specifically, the atrial-to-His bundle (A-H) interval was measured. Studies with the NS5A inhibitor ledipasvir could not be completed under the experimental conditions due to poor solubility
Journal of Hepatology 2016 vol. 64 | S425–S630
Background and Aims: The importance of the intestinal microbiota for the onset and clinical course of many diseases, including liver diseases like NASH and liver cirrhosis, is increasingly recognized. However, the role of intestinal microbiota in chronic HCV infection remains unclear. Methods: In a cross sectional approach the intestinal microbiota of 95 patients chronically infected with HCV (n = 57 w/o cirrhosis (NOCIR); n = 38 with cirrhosis (CIR) and 50 healthy controls (HC) was analysed. The V1-2 region of the 16S rRNA gene was amplified followed by sequencing on the Illumina MiSeq platform. Random subsampling was performed to achieve equal number of reads.
Methods: Patients with chronic hepatitis C without any antiviral therapy in the previous one year were included. Subjects with decompensated cirrhosis, hepatitis B o HIV coinfection, or previous diagnosis of diabetes mellitus were excluded. In 76 patients, we performed a blood laboratory test which include routine, metabolic and inflammatory parameters (retinol, retinol-binding protein 4, 25OH vitamin D, Vitamin E, lipopolysaccharide-binding protein, interleukin-6 and cystatin C). Insulin resistance was established with a HOMA value higher than 3. We determined single nucleotide polymorphisms rs7041 and rs4588 GC-DBP (Group specific component-Vitamin D-binding protein), rs 738409 PNPLA3 ( patatin-like phospholipase domain containing 3), and rs12979860 IL28B (interleukin-28 B) genes. Viral parameters like genotype o viral load were obtained and fibrosis staging was assessed with liver biopsy or transient elastography. Results: After backward logistic regression analysis, independent variables associated with insulin resistance were: 1) Gc1s/Gc1S Vitamin D-binding protein phenotype, that results from the homozygous carriage of the rs7041G/rs4588C haplotype ( p = 0.033); 2) low retinol/RBP4 ratio, reflecting a greater rate of unbound retinol-binding protein 4 ( p = 0.005); 3) older age ( p = 0.01); 4) high serum tryglicerides ( p = 0.026), and 5) advanced (F3F4) fibrosis stage ( p = 0.034). The AUROC provided by the multivariate model was 0.950 (95 % IC = 0.906–0.993). Conclusions: In addition to previously known ones, the Gc1s/Gc1s phenotype variant of vitamin D-binding protein and the unbound fraction of plasma retinol-binding protein 4 may be considered as factors related with the incidence, and possibly the risk, of insulin resistance in chronic hepatitis C patients. FRI-159 PATIENTS WITH CHRONIC HEPATITIS C CAN BE IDENTIFIED BY THEIR GUT MICROBIOTA B. Heidrich1,2,3, M. Vital2, I. Plumeier2, S. Kahl2, J. Kirschner1, L. Sollik1, S. Ziegert1, M. Manns1,3, H. Wedemeyer1,3, D. Pieper2,3. 1Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Hannover; 2Microbial Interactions and Processes Research Group, Center for Infection Research, Braunschweig; 3German Center for Infection Research (DZIF), Hannover-Braunschweig, Germany E-mail: [email protected]
Results: The number of phylotypes (S) was significantly lower in NOCIR vs. HC (386 ± 77 vs. 420 ± 72; p = 0.003) and CIR vs. HC (329 ± 86 vs. 386 ± 77; p = 0.009). Shannon diversity index H’ was different between CIR vs. HC (4.0 ± 0.5 vs. 4.4 ± 0.3; p < 0.001) and CIR vs. NOCIR (4.0 ± 0.5 vs. 4.4 ± 0.4; p = 0.002) while no differences were
Journal of Hepatology 2016 vol. 64 | S425–S630
S609
POSTER PRESENTATIONS observed between HC vs NO-CIR. S and H’ correlated negatively with ARFI (S: R = −0.590; p < 0.001; H’: R = −0.528; p = 0.002) in patients. The presence or absence of 14 genera, which were more often observed in either HC or NO-CIR was used to develop a score to differentiate between HC and NO-CIR. If a genus, that was more often observed in HC, was observed in a sample, it was scored + 1 whereas it was scored −1, when the genus was more often observed in NO-CIR. The total score ranged from −6 to +8. Assuming samples of a score >2 as being from HC, the score has an AUC of 0.917 with a sensitivity of 0.82, specificity of 0.92, PPV of 0.92 and NPV of 0.82. Additionally, we observed four distinct patterns in the relative abundance of genera. The relative abundance of 5 genera increased from HC to NO-CIR to CIR ( pattern 1), whereas the relative abundance of 8 genera decreased, and the abundance of those genera can, thus, be associated with fibrosis progression. Six genera showed a significant difference in abundance in CIR but comparable abundancies in HC and NO-CIR ( pattern 3), and their abundance could, thus be associated with cirrhosis. A further 6 genera showed a significant difference between HC and NO-CIR ( pattern 4), thus, being associated with HCV infection (Fig). Conclusions: These results demonstrate an association of chronic HCV infection with the community structure in the gut compared to HC even in the absence of cirrhosis. It has to be clarified if these differences in the microbiota are reversible with the clearance of HCV by novel DAAs. FRI-160 RISKS OF LIVER COMPLICATIONS AFTER A LONG-TERM FOLLOW-UP IN PATIENTS WITH CHRONIC HEPATITIS C WITH SUSTAINED VIROLOGIC RESPONSE B. Kutala1, M.-P. Ripault1, E. Estrabaud1, F. Khelifa-Mouri1, R. Nait1, N. Boyer1, N. Giuily1, C. Castelnau1, T. Asselah1, P. Marcellin1. 1Service d’hépatologie, Hôpital Beaujon, Clichy, France E-mail: [email protected] Background and Aims: The long-term effect of sustained virologic response (SVR) to antiviral therapy on the risk of developing hepatocellular carcinoma (HCC), liver decompensation, liver-related death, and overall death, in patients with chronic hepatitis C (HCV), has been determined by few studies in the short term and never in the long term. Methods: The study was conducted on 1024 patients with chronic hepatitis C treated with antiviral regimens based interferon (IFN) who had achieved SVR at the university center of Beaujon from 2000 to 2011. Complementary follow-up data were obtained by contacting patients themselves, their primary care physician, their municipality of birth and the French biomedicine agency for patients on list of liver transplantation. Results: The baseline median (IQR) age was 51 (40–59) years and 734 patients (72%) were men. The advanced fibrosis score (Metavir F3–F4) was in 403 patients (39%), genotype 1 found in 542 patients (53%). During the follow-up (mean of 5.6 years), among patients with advanced fibrosis (F3–F4), 19 patients died or underwent liver transplantation (LT), 31 patients developed HCC 14 patients had liver decompensation. However in patients without advanced fibrosis (F0–F2) 3, 2 and 2 patients respectively died or underwent LT, developed HCC and had liver decompensation. The incidences of overall death or liver transplantation, HCC and any liver decompensation per 100 person-years were significantly higher in F3-F4 patient time with 0.8, 1.3 and 0.6, compared to 0.2, 0.1, and 0.1 in patients without advanced fibrosis (F0-F2). Conclusions: Among patients with SVR, the risk of death or LT, HCC and liver decompensation were markedly reduced after 3 years follow-up the SVR. However, a long-term risk of developing HCC persists in patients with advanced fibrosis. There was no recurrence of HCV infection or evidence of liver disease progression in patients without advanced fibrosis (F0–F2). Future studies focusing on the
S610
better identification of patients with a long-term risk for HCC are needed. FRI-161 LONG-TERM ASSESSMENT OF RENAL FUNCTION IN PATIENTS FOLLOWING ANTIVIRAL THERAPY FOR CHRONIC HEPATITIS C VIRUS INFECTION C.-H. Liu1, C.-L. Chen2, C.-J. Liu2, T.-H. Su1, H.-C. Yang1, P.-J. Chen2, D.-S. Chen3, J.-H. Kao3. 1Internal Medicine; 2Graduate Institute of Clinical Medicine; 3Hepatitis Research Center, National Taiwan University Hospital, Taipei, Taiwan E-mail: [email protected] Background and Aims: The long-term progression of renal function in patients following antiviral therapy for chronic hepatitis C virus (HCV) infection remains unclear. Methods: A total of 1,569 HCV-infected patients receiving peginterferon/ribavirin or direct acting antiviral agents (DAAs) were recruited in this prospective cohort study. Serial renal function assessment was evaluated by modification of diet in renal disease equation (MDRD). Linear mixed-effect models were used to estimate the effect of sustained virologic response (SVR) on the changes of MDRD during follow up. Adjusted variables included age, sex, history of DM, hypertension and hyperlipidemia and the interaction between time and SVR. Results: The median time (interquartile range) of post-treatment follow-up was 48 (6–66) months and the SVR rate was 80.1%. The analysis was restricted to 48 months of follow-up. The renal function was similar at the beginning of follow up, regardless of SVR status (82.7 ± 21.4 vs. 80.1 ± 21.8 for SVR vs. non SVR), but significantly lower among patients with DM and hypertension. As time progress, the MDRD started to increase with an average of 0.21 (95% CI = 0.17–0.26) per 6 month. The slope of the increase was higher for those not achieving SVR (averaging 0.25 increase per 6 month, adjusted p for trend <0.001) than those achieving SVR (averaging 0.07 per 6 month, adjusted p for trend = 0.054). Conclusions: In patients undergoing antiviral therapy for HCV, those who achieved SVR was associated with slower progressive loss of renal function. FRI-162 EFFECT OF AMIODARONE AND HCV DIRECT-ACTING ANTIVIRAL AGENTS ON CARDIAC CONDUCTION IN NONCLINICAL STUDIES G. Liu1, C.H. Tay1, S. Rajamani1, A. Ray1, L. Stamm1, J. Vick2, M. Wilichinsky1, J. McHutchison1, D. Brainard1. 1Gilead Sciences, Inc., Foster City; 2ChanTest, a Charles River Company, Ohio, United States E-mail: [email protected] Background and Aims: In the post-market setting, serious cases of symptomatic bradycardia have been reported in patients taking amiodarone with Sovaldi in combination with an HCV direct-acting antiviral (DAA). The underlying mechanism for these events is unknown but is currently under investigation. Amiodarone, a multiion channel inhibitor including Ca2+ currents, has been shown to affect the atrial-to-ventricular conduction precipitating bradycardia. Sofosbuvir, a HCV NS5B polymerase nucleotide inhibitor, did not alter the heart rate, blood pressure, or EKG in telemeterized dogs and up to supratherapeutic doses in a clinical TQT study. The effect of amiodarone and DAAs, alone and in combination, on cardiac conduction in guinea pig isolated hearts, and in vitro inhibition of human calcium channels were determined. Methods: Using guinea-pig isolated hearts, the effect of amiodarone, daclatasvir (a NS5A inhibitor), simeprevir (a NS3/4A protease inhibitor), sofosbuvir and GS-331007 (nucleoside analogue of sofosbuvir) alone or in combination at their maximal concentrations achieved in plasma, on cardiac conduction was determined; specifically, the atrial-to-His bundle (A-H) interval was measured. Studies with the NS5A inhibitor ledipasvir could not be completed under the experimental conditions due to poor solubility
Journal of Hepatology 2016 vol. 64 | S425–S630