Circulating serum exosomal microRNA-203 as a non-invasive biomarker for predicting prognosis in hepatocellular carcinoma

POSTER PRESENTATIONS SAT-491 Serum vascular cell adhesion molecule-1 predicts significant liver fibrosis in obese patients with non-alcoholic fatty liver disease S. Lefere1, F. Van de Velde2, L. Devisscher1, M. Bekaert2, S. Raevens1, X. Verhelst1, Y. Van Nieuwenhove3, M. Praet4, A. Hoorens4, C. Van Steenkiste1, H. Van Vlierberghe5, B. Lapauw2, A. Geerts1. 1 Gastroenterology and Hepatology; 2Endocrinology; 3Gastroinestinal Surgery; 4Pathology, Ghent University Hospital, Ghent; 5 Gastroenterology and Hepatology, Ghent University Hospital, Gent, Belgium E-mail: [email protected] Background and Aims: Non-alcoholic fatty liver disease (NAFLD) is the most common chronic liver disease worldwide and is strongly associated with obesity, dyslipidemia and insulin resistance. NAFLD often presents as simple steatosis (NAFL) but can progress to nonalcoholic steatohepatitis (NASH) and fibrosis. Current non-invasive biomarkers are not tailored to identify significant (≥F2) fibrosis, although recent guidelines recommend a stringent follow-up of this patient population. We and others have reported on the role of pathological angiogenesis in the pathogenesis of NAFLD, highlighting pro-angiogenic factors as potential diagnostic markers. We aimed to investigate the applicability of angiogenic factors as non-invasive diagnostic tools for NASH-associated fibrosis in obese patients. Methods: Serum protein levels and visceral adipose tissue gene expression of endothelial dysfunction and angiogenic markers were analyzed by multiplex bead-based assay and quantitative RT-PCR, respectively, in sixty-one morbidly obese male patients undergoing bariatric surgery and in thirty-five control patients. Results: We identified serum vascular cell adhesion molecule-1 (VCAM-1) as an independent predictor for ≥F2 fibrosis (median 14.0 vs. 8.7 ng/mL in patients with and without significant fibrosis; P < 0.0001) with an area under the receiver operating characteristics curve (AUROC) of 0.80. The cut-off point of 13.2 ng/mL showed a sensitivity of 80% and specificity of 83%. The AUROC increased to 0.86 when VCAM-1 and the presence of type 2 diabetes were combined. These AUROCs were higher than those for the simple fibrosis risk scores FIB-4 and BAAT. In line with these results, VCAM-1 visceral adipose tissue gene expression was also elevated in patients with fibrosis (P = 0.030). Conclusions: Serum VCAM-1 levels were able to accurately predict significant (≥F2) fibrosis in obese men with NAFLD. SAT-492 The assessment of liver fibrosis stage by two-dimensional shear wave ultrasound and transient elastography in patients with chronic liver disease S.G. Kim1, B. Lee2, J.J. Yoo1, Y.S. Kim1, S.W. Jeong3, J. Youn, Jang3, S.H. Lee4, Y.D. Kim5, G.J. Cheon5, H.S. Kim4, B.S. Kim1. 1Soonchunhyang University Bucheon Hospital, School of Medicine; 2Biostatistical Consulting Unit, Soonchunhyang University Bucheon Hospital, Bucheon; 3 Soonchunhyang University Bucheon Hospital, School of Medicine, Seoul; 4Soonchunhyang University Bucheon Hospital, School of Medicine, Cheonan; 5Gangneung Asan Hospital, Gangneung, Korea, South E-mail: [email protected] Background and Aims: Several real-time two-dimensional shear wave elastography (2D-SWE) have been developed to assess liver fibrosis with readily use of combining elastography and traditional ultrasound imaging. However, compared with transient elastography (fibroscan), the diagnostic accuracy and clinical usefulness of these methods were not fully validated. In this study, newly developed 2DSWE (LOGIQ E9, GE healthcare, UK) was evaluated for predicting liver fibrosis stage and compared with fibroscan. Methods: Out of 2,144 patients who received 2D-SWE during May 2015 to Nov 2016, one hundred-forty (6.5%) who failed to get available value of 2D-SWE due to obesity or poor echo window and S672

207 (9.7%) with high value of AST or ALT were excluded in the analysis. Liver biopsy was performed in 244 patients. 2D-SWE measurement was considered valid when homogenous color pattern in a region of interest of at least 10 mm was shown at 10 different sites. Diagnostic performance was calculated using area under the receiver operating characteristics curve (AUROC). Results: Patients were male predominant (53.7%), their mean age was 49.1 ± 13.9 years old and most common etiology of liver disease was hepatitis B (34.4%) followed by autoimmune hepatitis (17.6%). Liver fibrosis stage consisted of F0 (14.8%), F1 (20.1%), F2 (23.8%), F3 (15.6%) and F4 (25.8%). Overall, 2D-SWE was well correlated with transient elastography (r = 0.855, P < 0.001). 2D-SWE median values (kPa) increased with increasing stage of liver fibrosis [ F0 (5.0 ± 1.0), F1 (5.7 ± 1.2), F2 (6.8 ± 2.0), F3 (8.7 ± 2.1), F4 (12.5 ± 2.9)] ( p for trend <0.001). For the diagnosis of liver cirrhosis, AUROCs and optimal cutoff of 2D-SWE were 0.931 (95% confidence interval [CI], 0.905– 0.958) and 9.9 kPa which was not significantly different from fibroscan [0.918 (95% CI 0.878–0.957)] ( p = 0.186). The sensitivity, specificity, positive predictive value and negative predictive value of 2D-SWE for predicting cirrhosis were 90.2%, 86.5%, 70.7% and 94.8% respectively. For diagnosing significant liver fibrosis (≥F2), AUROCs and optimal cutoff of 2D-SWE were 0.889 (95% CI, 0.849–0.929) and 6.68 kPa.

Conclusions: With good comparability to fibroscan and availability of a conventional ultrasound examination, Two-dimensional SWE is an useful tool for stratifying liver fibrosis stage and diagnosing liver cirrhosis. SAT-493 Circulating serum exosomal microRNA-203 as a non-invasive biomarker for predicting prognosis in hepatocellular carcinoma S.Y. Jang1, G. Kim2, K. Hur2, S.Y. Park1, W.Y. Tak1, Y.O. Kweon1, Y.R. Lee1, J.S. Yoon1, S.H. Kwon1, H.S. Kim1, J.S. Lee1, J.G. Park3. 1Internal Medicine, Kyungpook National University Hospital; 2Biochemistry and Cell Biology, Kyungpook National University School of Medicine; 3Internal Medicine, Yeungnam University Medical Center, Daegu, Korea, South E-mail: [email protected] Background and Aims: Recently, cancer cell-derived extracellular vesicles have been known to contain various intracellular biomolecules including microRNAs (miRNAs). The aim of this study was to evaluate prognostic value of serum exosomal miRNAs serving as a non-invasive biomarker in hepatocellular carcinoma (HCC). Methods: We isolated exosomes from serum samples of 102 HCC patients by ultracentrifugation and exosomes were confirmed by expression of exosome markers (CD9, CD63, ALIX, and TSG101) based on immunoblotting. We analyzed the expression of 6 miRNAs (miRNA-24, -130a, -182, -203, -373, and -423) and investigated expression of the 6 miRNAs in HCC tissues and paired liver tissues. MiRNAs expression was determined by quantitative real-time PCR and normalized by cel-miR-39 and RNU6B expression for serum and tissue samples, respectively. The chi-square or Fischer exact test was used to analyze the relationship between miRNA expression level and clinicopathological characteristics. Overall and disease

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POSTER PRESENTATIONS progression-free survival were plotted as Kaplan-Meier curves and the log-rank test was used for evaluating differences in survivals, respectively. Results: We observed that the level of miRNAs (miRNA-24, -130a, -182, -203, and -373) from serum exosome were elevated in HCC patients comparing to that of normal healthy controls. In further comparison between early stage and advanced stage of HCC patients, serum exosomal miRNA-203 (P < 0.05) and miRNA-373 (P < 0.05) were significantly up-regulated in advanced HCC patients. Among age, sex, Child-Pugh class, TNM stage, α-fetoprotein, serum-exosomal miR-203, and serum-exosomal miR-373, TNM stage (IV) (hazard ratio [HR] = 19.147, 95% confidence interval [CI] = 4.054–90.430, p = 0.0002) and high serum-exosomal miR-203 (HR = 3.788, 95% CI = 1.223–11.734, p = 0.0210) were prognostic factors for overall survival and TNM stage (III) (HR = 3.913, 95% CI = 1.135–13.489, p = 0.0307), TNM stage(IV) (HR = 15.832, 95% CI = 5.016–49.970, p < 0.0001) and high serum-exosomal miR-203 (HR = 2.503, 95% CI = 1.075–5.826, p = 0.0333) were prognostic factors for disease progression-free survival in multivariate analysis.

Conclusions: We provided the evidence of prognostic value of serum exosomal miR-203 as a potential non-invasive biomarker in HCC patients. SAT-495 Community Approach Targeting Cirrhosis and Hepatocellular carcinoma (CATCH): community prevalence of advanced fibrosis in viral hepatitis S. Bloom1,2,3, W. Kemp3,4, A. Dev4,5, P. Gow6,7, A. Nicoll1,4,7, S. Roberts3,4, S. Bell7,8, I. Kronborg7,9, V. Knight5, S. Sood10, D. Lewis1,2, J. Lubel1,2 and Melbourne Liver Group. 1Gastroenterology and Hepatology, Eastern Health; 2Eastern Health Clinical School, Monash University; 3Gastroenterology, Alfred Health; 4Monash University; 5 Gastroenterology, Monash Health; 6Gastroenterology and Hepatology, Austin Health; 7University of Melbourne; 8Gastroenterology, St Vincent’s Hospital; 9Gastroenterology, Western Health; 10Gastroenterology and Hepatology, Royal Melbourne Hospital, Melbourne, Australia E-mail: [email protected] Background and Aims: The majority of chronic hepatitis C (CHC) and chronic hepatitis B (CHB) is managed within primary care and the prevalence of advanced fibrosis in this group is unknown. This study aims to estimate the prevalence of significant fibrosis and cirrhosis as assessed by LSM in an at-risk population and explore the role of this investigation in facilitating the detection of hepatocellular carcinoma (HCC).

Methods: Participants were recruited from 21 primary care practices across Melbourne, Australia. Inclusion criteria included age >18 yrs, CHB or CHC duration >6 months, no prior or recent (<18 mths) specialist input and no history of HCC. Clinical assessment, LSM (Fibroscan® 402) and blood analysis were performed in the primary care setting. Scans were considered reliable if the success rate was >60% and IQR/median stiffness <0.3. LSMs of 8.0 kPa and 12.5 kPa were taken as cut-offs to represent significant fibrosis (>F2) and cirrhosis (F4) respectively. Referral bias was assessed using a consecutively recruited hospital control group (h) undergoing identical assessment to the community group (c). Results: Over 24months, 1,523 were invited to participate of which 1,043 participants were assessed (753 cCHC/290 cCHB). LSM failure occurred in two (0.2%) subjects and phlebotomy failure in seven (0.7%). The M probe was used in 95.4% and the XL probe in 4.6%. The mean LSM was 9.9 kPa in cCHC and 5.0 kPa in cCHB with no difference in mean LSM between the cohorts (CHC p = 0.92/CHB p = 0.17). An LSM ≥ 8kPa was observed in 31.2% (cCHC 40.6%/cCHB 7.2% p < 0.01) and a LSM ≥12.5 kPa in 11.6% (cCHC 15.9%/cCHB 0.6% p < 0.01). Three additional cCHB patients were diagnosed as cirrhotic on clinical grounds (all LSM > 11.0), with a final prevalence of 2.8%. (vs. 5.0% hCHB p = 0.04). The prevalence of LSM ≥ 12.5 kPa was no different between cCHC and hCHC (15.9% vs. 21.2% p = 0.06). On multivariate analysis; advanced age (OR1.048 p < 0.01), waist circumference (OR1.052 p < 0.01) and at risk alcohol use (>140 g/ week, OR1.937 p < 0.01) were associated with an LSM ≥ 12.5 kPa in cCHC. In cCHB only viral load (OR4.597 p = 0.03) was predictive of cirrhosis on multivariate analysis. Four HCC (all BCLC A) were detected with an incidence rate of 3.9% in our cirrhotic patients. Conclusions: Based on LSM, the prevalence of advanced liver fibrosis in community CHC is comparable to a hospital cohort. This data highlights the utility of LSM to identify those at risk in the community for liver related events and the need for community-based surveillance programs for HCC. SAT-496 Transient Elastography has limited efficacy for fibrosis assessment in End Stage Renal Disease patients on maintenance haemodialysis with suspected liver disease S. Taneja1, A. Borkakoty1, S. Rathi1, A. Duseja1, R.K. Dhiman1, Y. Chawla1. 1Hepatology, PGIMER, Chandigarh, India E-mail: [email protected] Background and Aims: Patients with End Stage Renal Disease (ESRD) are at high risk of acquiring chronic viral hepatitis B and C and subsequent liver disease. While non-invasive assessment of liver fibrosis by Transient Elastography has emerged as a reliable tool, its validity in patients with volume overload states like renal failure and heart failure is ambiguous. Aims To study the correlation of liver stiffness measurement (LSM) with liver fibrosis on histology, and change in LSM and body impedance parameters before and after hemodialysis (HD) in ESRD patients with suspected liver disease. Methods: We prospectively enrolled 68 patients of ESRD on maintenance hemodialysis (MHD) with suspected liver disease and compared LSM before and after HD. The change in LSM (DLSM) values were then correlated with the change in body weight and total body water, duration and frequency of HD. 18 patients underwent a liver biopsy, and correlation of LSM with histopathological grade of fibrosis was assessed. Results: The mean age of the study population was 40 ± 14 yrs. Hepatitis C was the most common cause of liver disease followed by hepatitis B. There was a significant reduction of LSM values after HD (18.5 ± 14.9 vs. 11.2 ± 10.1 kPa, P < 0.001), with a mean LSM reduction of 7.2 ± 8.2 kPa. The decline in LSM after HD strongly correlated with the LSM at baseline (r = 0.77, P = <0.001). The population was divided in four quartiles based on LSM, which revealed lowest decline in

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