The HCV-Associated Hepatocarcinogenesis in Intracellular Low Viral Load Cells

( Chia-yen Dai ),( Shu-chi Wang ),( Chung-feng Huang ),( Wang-long Chung ),( Ming-lung Yu ) 대한간학회 2018 춘·추계 학술대회 (KASL) Vol.2018 No.1

Aims: There are differential viral load distribution in HCV infected liver tissues. We conduct the present study aimed to dissect the different viral load cells to investigate the viral-host resistance on the HCV asssoicated hepatocarcinogenesis. Methods: The study was performed using a replicated in vitro HCV-fluorescence infection model to discuss HCV-high viral load (HVL) cells and the HCV-low viral load (LVL) cells by Flow sort system. The next generation RNA sequence and miRNA array were used to explore the gene profiles and miRNA expression on different HCV-viral load cell populations. Results: The ROS indicator shows that the ROS abundantly in low viral load cells. The RNA-Seq analysis showed that significant enrichment in Cancer (P=5.00E-02 - 2.76E-04; 40 molecules) and a network of the Cellular Movement, Immune Cell Trafficking, Inflammatory Response (Score: 18) by IPA analysis. Protein analysis results confirmed the GADD45A and iNOS overexpression in the LVL cells which verified the oxidative stress qPCR array data in LVL cells. We also found the up-regulated Src oncoprotein expression and down-regulated E-cadherin expression in LVL cells. The miRNA array showed that miR-194, miR-192/215 and miR-10a were preferentially expressed in low viral load cells. Conclusions: With our established cell sorting system, this study provided the gene network between viral and host cells resistance by different viral load cells. The findings show the activated oxidative stress related-gene expression in hepatocytes is associated with the HCV-infected epithelial-to-mesenchymal transition (EMT), providing an important link between HCV viral load and liver cancer. The miRNA-gene intergraded dada need further studies.

Special Issue in Hepatitis C Virus Treatment: Hepatitis B Virus Reactivation and Occurrence and Recurrence of Hepatocellular Carcinoma

( Chia-yen Dai ) 대한간학회 2018 춘·추계 학술대회 (KASL) Vol.2018 No.1

The overall outcome of patients with chronic hepatitis C virus (HCV) infection may have the chance of improvement from the advancement in the treatment efficacy of antiviral therapies. With higher efficacy of the standard of care (pegylated interferon- alpha (PegIFN) plus ribavirin therapy and/or new all oral direct-acting antiviral agents (DAAs), the sustained virological response (SVR) has greatly improved the treatment efficacy. However, some special issues for some patient groups need to be noticed. For patients with hepatitis B virus (HBV)/HCV coinfection, hepatitis B reactivation associated with DAA therapy is emerging to be an important cause of morbidity and mortality. The positivity of HBsAg and HBV DNA are the major determinants of hepatitis B reactivation. With the previous reports by the IFN-based therapy, hepatitis B reactivation was observed about 40% during 5-year post-treatment follow-up with rare clinical hepatitis event. After treatment of DAAs for CHC, the hepatitis B reactivation seems different from the IFN-based therapy. Recent data suggest that in patients with positive HBsAg and HCV infection, reactivation of HBV can occur in up to 70% of the subjects during the DAA therapy and within one year after cessation of DAA therapy. Less than 20% of the subjects developed hepatitis flare accompanying HBV reactivation. The major guidelines have highlighted the needs of monitoring for HV reactivation or even initiating the prophylactic therapy with nucleos(t)ide analogues which may need further studies to clarify the predictive factors and the patient groups who will get the benefits. Another special consideration that should be discussed is related to hepatocellular carcinoma (HCC). The effect of the viral eradication on the development of HCC is an important issue which reflects the outcome of secondary and tertiary prevention in CHC patients with antiviral therapies. With the SVR achieved after the IFN-based therapy, it has been shown that successful antiviral therapy can reduce the risk of HCC development in HCV patients who were without HCC. For patients with HCC receiving HCC therapy, there were some reports revealing no benefit of eradication of HCV on the prevention of the recurrence of HCC. However, large cohort studies have shown that for patients after HCV-related HCC After curative therapy, IFN therapy has been shown to improve outcomes these patients. In the DAA era, with the very high cure rate of HCV, most of the reports show that DAAs therapy seems not to be associated with the occurrence of the HCC. There were some reports indicated the effect of secondary prevention of HCC in CHC patients. Nevertheless, the risk of HCC in patients after therapy for HCC, such as the curative therapy has been debated with DAA therapy. Some studies have shown the lack of preventing recurrence of the HCC in patients with DAAs therapy after treatment of the HCC. On the country, there is the raised concern of the surprisingly increased risk of recurrence of the HCC in these DAAs-treated patients after the curative therapies for HCC. There are more evidences recently reported that no observation about such an increasing of the recurrence of HCC. The efficacy of tertiary prevention needs further long-term studies and assessment.

The Different Expression of Gene Profiles on Hepatocellular Carcinoma Cells with Different Intracellular Hepatitis C Viral Load

( Chia-yen Dai ),( Shu-chi Wang ),( Meng-hsuan Hsieh ),( Cheng-fu Yang ),( Ching-i Huang ),( Chung-feng Huang ),( Ming-lun Yeh ),( Jee-fu Huang ),( Wang-long Chung ),( Ming-lung Yu ) 대한간학회 2017 춘·추계 학술대회 (KASL) Vol.2017 No.1

Aims: The different hepatitis C virus (HCV) replication has been reported among individual hepatocytes in chronic HCV infection by identifying hepatocytes with different HCV RNA levels. We have previously established a fluorescence-activated cell sorting (FACS) protocol to study the effects of different intracellular viral loads in HCV-infected cells. The present study aimed to further study the gene expression on different hepatocellular carcinoma (HCC) cells with different HCV viral load. Methods: The JFH1-EYFP viral florescence intensity was used to sort the high and low viral load cells after 5 days infection in vitro which has been shown in our previous study that infected cells efficiently and accurately discriminated between high- and low-viral load cell populations. The next generation sequence-RNA sequence was used to clarify the mRNA and miRNA gene network between HCV-high and HCV-low infected cells of the HCC cell line. Venn diagram summarizing the probe sets that were differentially expressingbetween the Huh7.5.1 versus each differential viral load cell population and miRDB and miRTar databases were used to predict HVL and LVL/S2 unique miRNA target genes. Results: By analyzing the NGS dataset and miRNA microarray dataset, of the significant transcripts, three miRNA were unique for the LVL/S2 cells and nine miRNA unique for the HVL. Twenty-three miRNA were common for all 3 viral load groups. We verified them by q-PCR and data confirmed the array data expression level. We found that high viral loads were associated with cell inflammation- and cell death-associated pathway; and the low viral loads were associated many stress response- and cell adhesion molecular (CAMs)-related genes. Conclusions: With the established cell sorting protocol, we have demonstrated that different gene network between HCV-high and HCV-low infected cells in JFH1-EYFP infectious cells exists. Our results may provide a boarder gene regulation map between high and low viral load cell populations.

Motivation of teaching marine leisure and tourism and students’ ocean literacy and engagement in marine related career plan

Cheng-Chieh Chang,Chia-Dai Yen,Lusi Victoria 한국호텔외식관광경영학회 2015 한국호텔외식경영학회 학술발표논문집 Vol.- No.-

Serial Serum MHC Class I Chain-Related a Levels in Predicting Hepatocellular Carcinoma in Chronic Hepatitis C Patients with Curative Antiviral Treatment

( Ming Lung Yu ),( Chung-feng Huang ),( Chia-yen Dai ),( Jee-fu Huang ),( Wan-long Chuang ) 대한간학회 2017 춘·추계 학술대회 (KASL) Vol.2017 No.1

Aims: MHC class I chain-related A (MICA) genetic variants and its serum level (sMICA) were associated with hepatitis C virus (HCV) related hepatocellular carcinoma (HCC) in untreated cohorts. The dynamic changes of serial sMICA levels regarding anti-HCV treatment and consequent HCC development is elusive. Methods: Single nucleotide polymorphism rs2596542 of MICA and serial sMICA levels were tested in chronic hepatitis C (CHC) patients with sustained virological response after antiviral treatment. Forty-two patients who developed HCC and another 84 age-, sex- and cirrhosis-propensity score matched non-HCC controls were compared. Serial sMICA levels were measured at three-time points: within 6 months of pretreatment (pre-sMICA), 6 months after the end of treatment (post-sMICA) and last visit before HCC occurrence or not (last-sMICA). Results: Compared to patients without HCC occurrence, those with HCC had lower platelet counts, higher levels of post-sMICA (197.4+398.0 pg/mL vs. 57.6+89.6 pg/mL, P=0.03) and last-sMICA (320.4+508.4 pg/mL vs. 37.7+140.2 pg/mL, P<0.001). Cox regression analysis revealed that last-sMICA is the only factor predictive of HCC development (hazard ratio [HR]/ 95 % confidence intervals [CI.]: 2.27 (per 1 log pg/mL increase)/1.672-3.082, P<0.001). Patients without HCC had a significantly decreased trend of sMICA levels during follow-up (trend P=0.001). In contrast, HCC patients had an increased trend of sMICA levels (trend P=0.024). MICA rs2596542 GG genotype carriers without HCC had a significantly decreased trend of sMICA levels during follow-up (trend P<0.001). However, HCC patients who carried GG genotype had a substantially increased trend of sMICA levels (trend P=0.06). Nevertheless, both trends were not observed in A allele carriers with or without HCC development. t three-time points: within 6 months of pretreatment (pre-sMICA), 6 months after the end of treatment (post-sMICA) and last visit before HCC occurrence or not (last-sMICA). Conclusions: Serial sMICA levels could serve as a surrogate marker for HCC development in CHC patients with SVR. The clinical utility is restricted to MICA rs2596542 GG genotype carriers. CA (320.4+508.4 pg/mL vs. 37.7+140.2 pg/mL, P<0.001). Cox regression analysis revealed that last-sMICA is the only factor predictive of HCC development (hazard ratio [HR]/ 95 % confidence intervals [CI.]: 2.27 (per 1 log pg/mL increase)/1.672-3.082, P<0.001). Patients without HCC had a significantly decreased trend of sMICA levels during follow-up (trend P=0.001). In contrast, HCC patients had an increased trend of sMICA levels (trend P=0.024). MICA rs2596542 GG genotype carriers without HCC had a significantly decreased trend of sMICA levels during follow-up (trend P<0.001). However, HCC patients who carried GG genotype had a substantially increased trend of sMICA levels (trend P=0.06). Nevertheless, both trends were not observed in A allele carriers with or without HCC development. t three-time points: within 6 months of pretreatment (pre-sMICA), 6 months after the end of treatment (post-sMICA) and last visit before HCC occurrence or not (last-sMICA).

Achievement Goal Orientation in Chinese Societies: Implications for College Counseling

Ching-Chen Chen,Wei-Wen Chen,George B. Richardson,Chia-Laing Dai,Jared Lau,Danica G. Hays 한국상담학회 2018 Journal of Asia Pacific counseling Vol.8 No.1

Achievement goal orientation has been linked to various academic and psychosocial outcomes within the United States and globally. As college counselors increasingly work with a diversified student body, it is imperative that they understand goal orientation and how it should be assessed among various cultures and subcultures. This study evaluated the functioning of the Achievement Goal Scale items across samples from Taiwan, Hong Kong, and Macau and tested whether metric (i.e., goal orientation dimensions had the same meanings) and scalar (i.e., means could be compared across groups) invariance held. Findings suggest that goal orientation can be assessed across Chinese societies and, therefore, used to inform the development of culturally appropriate counseling approaches when working with college students. Implications for counseling practice and research are provided.

Time-degenerative Factors and the Risk of Hepatocellular Carcinoma after Antiviral Therapy among HCV Patients: A Model for Prioritization of Treatment

( Ming-lung Yu ),( Chung-feng Huang ),( Ming-lun Yeh ),( Jee-fu Huang ),( Chia-yen Dai ),( Wan-long Chuang ) 대한간학회 2016 춘·추계 학술대회 (KASL) Vol.2016 No.1

Aims: Age and hepatic fibrosis are the factors that increase the risk of hepatocellular carcinoma (HCC) over time. We aimed to explore their impac at the initiation of antiviral therapy on HCC among chronic hepatitis C (CHC) patients. Methods: A total of 1281 biopsy-proven CHC patients receiving interferon- based therapy were followed for a mean period of 5.5 years. Results: The 5-year cumulative incidence of HCC did not differ between non-SVR and SVR patients who were <40 years old (7.7 % vs. 0.5%, P=0.1), but was significantly higher in non-SVR patients between 40 and 55 years old (18.0% vs. 1.3%, P<0.001) and >55 years old (15.1% vs. 7.9%, P=0.03). Compared with SVR, non-SVR was independently predictive of HCC in patients 40-55 years old (hazard ratio [HR]/95% confidence intervals [CI]: 10.92/3.78-31.56, P<0.001) and >55 years old (HR/CI: 1.96/1.06-3.63, P=0.03) but not in patients <40 years old (HR/CI: 2.76/0.41-18.84, P=0.3). The 5-year cumulative incidence of HCC did not differ between non-SVR and SVR patients whose fibrosis stage was F0-1 (4.6% vs. 1.9%, P=0.25) but was higher in non-SVR patients with F2-3 (21.4% vs. 4.3%, P<0.001) or F4 (33.5% vs. 8.4%, P=0.002). Compared with SVR, non-SVR was independently predictive of HCC in patients with F2-3 (HR/CI: 4.36 /2.10-9.03, P<0.001) and F4 (HR/CI: 3.84/1.59-9.30, P=0.03) but not in those with F0-1 (HR/CI: 1.53/ 0.49-4.74, P=0.47). Conclusions: Delayed HCV clearance for patients with CHC > 40 years old or with a fibrosis stage > 2 increases the risk of HCC over time.

Significant down-regulation of growth hormone receptor expression revealed as a new unfavorable prognostic factor in hepatitis C virus-related hepatocellular carcinoma

( Ching-chih Lin ),( Ta-wei Liu ),( Ming-lun Yeh ),( Yi-shan Tsai ),( Pei-chien Tsai ),( Chung-feng Huang ),( Jee-fu Huang ),( Wan-long Chuang ),( Chia-yen Dai ),( Ming-lung Yu ) 대한간학회 2021 Clinical and Molecular Hepatology(대한간학회지) Vol.27 No.2

Background/Aims: Growth hormone (GH) is the main regulator of somatic growth, metabolism, and gender dimorphism in the liver. GH receptor (GHR) signaling in cancer is derived from a large body of evidence, although the GHR signaling pathway involved in the prognosis of hepatocellular carcinoma (HCC) in patients with hepatitis C virus (HCV)-related HCC, remains unclear. We aimed to explore the expression of GHR and analyze its association with clinicopathologic features and prognosis of patients with chronic hepatitis C and HCC. Methods: The expression of GHR mRNA was investigated by quantitative real-time polymerase chain reaction in paired tumors and adjacent non-tumorous (ANT) liver tissues of 200 patients with chronic hepatitis C and HCC. Western blotting and immunofluorescence assays using the HCV-infected Huh7.5.1 cell model was performed. Results: GHR mRNA was significantly lower in HCV-HCC tissues than in corresponding ANT liver tissues. GHR mRNA and protein levels also decreased in the HCV-infected Huh7.5.1 cell model. Notably, lower GHR expression was associated with age of >60 years (P=0.0111) and worse clinicopathologic characteristics, including alpha-fetoprotein >100 ng/mL (P=0.0403), cirrhosis (P=0.0075), vascular invasion (P=0.0052), pathological stage II-IV (P=0.0002), and albumin ≤4.0 g/dL (P=0.0055), which were linked with poor prognosis of HCC. Most importantly, the high incidence of recurrence and poor survival rates in patients with a low ratio of tumor/ANT GHR (≤0.1) were observed, indicating that low expression levels of GHR had great risk for development of HCC in patients with chronic hepatitis C. Conclusions: Our study demonstrates a significant down-regulation of GHR expression as a new unfavorable independent prognostic factor in patients with chronic hepatitis C and HCC. (Clin Mol Hepatol 2021;27:313-328)

Response to DAA in HCV Patients with HCC from East Asia: A REAL-C Study with Propensity Score Matching (PSM)

( Mindie H. Nguyen ),( Norihiro Furusyo ),( Dae Won Jun ),( Ming-Lung Yu ),( Jia-Horng Kao ),( Masaru Enomoto ),( Eiichi Ogawa ),( Etsuko Ilio ),( Chen-Hua Liu ),( Akihiro Tamori ),( Chia-Yen Dai ),( 대한간학회 2018 춘·추계 학술대회 (KASL) Vol.2018 No.1

Aims: Reports suggest HCV-HCC patients do not respond as well to the IFN-free DAAs, but background risks and confounders for treatment failures may not have been adequately controlled. Our goal was to compare SVR12 of DAAs in East Asian patients with HCV-HCC to those without HCC using PSM to balance the HCC and non-HCC groups. Methods: Data were from 10 study centers comprising of 30 clinical sites in Hong Kong, Japan, Korea, and Taiwan representing the Real-World Evidence from the Asia Liver Consortium for Chronic Hepatitis C (REAL-C) - a registry of patients treated with IFN-free DAAs in routine practice (n=3702). 1:1 PSM matching on cirrhosis, prior treatment, baseline platelet, age, sex, baseline HCV RNA, treatment regimen, baseline ALT, HCV genotype, and BMI was used to balance the groups at baseline. Results: In our cohort, there were 195 patients with HCC at baseline or prior to DAA initiation and 3507 patients who did not have HCC at baseline. Prior to PSM, HCC patients were significantly older, more likely male, more likely to have renal insufficiency, cirrhosis, and decompensation (all P< 0.004). After PSM, there were 171 HCC and N=171 non-HCC patients for analysis. As shown in Table 1, there were no significant differences in the baseline characteristics between the matched HCC and non-HCC cohorts. The majority (51-55%) of both groups received LDV/SOF; eight (three HCC, five non-HCC) stopped treatment before completion while ~10-12% had an adverse reaction (most common: anemia [ >~5-6%] and fatigue [~3-5%]). There were seven deaths: five in the HCC group (four were liver-related) and two in the non-HCC group (both were non-liver-related). Overall, SVR12 rate was >96% for both groups with no significant differences. (Table 2) Conclusions: This PSM study compared treatment for HCV patients with/without HCC, finding no difference in treatment tolerability, completion, and cure rates.

Real-World Sustained Virologic Response Rates (SVR12) with Interferon (IFN)-Free Direct Acting Antiviral (DAA) Therapy in East Asia- Results from REAL-C (Real-World Effectiveness from the Asia Liver Consortium for Chronic Hepatitis C)

( Mindie H. Nguyen ),( Norihiro Furusyo ),( Dae Won Jun ),( Ming-Lung Yu ),( Jia-Horng Kao ),( Masaru Enomoto ),( Eiichi Ogawa ),( Etsuko Ilio ),( Chen-Hua Liu ),( Akihiro Tamori ),( Chia-Yen Dai ),( 대한간학회 2018 춘·추계 학술대회 (KASL) Vol.2018 No.1

Aims: Since their recent introduction in Asia, IFN-free DAAs have revolutionized treatment of chronic hepatitis C across all HCV genotypes. However, experience from large and diverse routine clinical practice is still limited. The aim of this study was to report real-world outcomes from a large multinational co hort of East Asian HCV patients treated with IFN-free DAAs. Methods: Data were obtained using a required case report form from the REAL-C registry of patients who were initiated on IFN-free DAA therapy in routine practice and represented 10 study centers inclusive of 30 clinical sites in Hong Kong, Japan, Korea, and Taiwan. Cirrhosis was determined by liver biopsy, noninvasive tests (elastography/fibroscan, fibrotest), or the presence of clinical, radiologic, endoscopic, laboratory evidence of cirrhosis and/or portal hypertension. Results: A total of 3702 patients have been registered. Table 1 displays the patient characteristics. The average age was 63.6±12.8; 17.7% had diabetes, 8.7% had chronic renal impairment, 26% had cirrhosis (5.1% decompensated cirrhosis), and 5.4% had HCC at baseline or prior to DAA treatment initiation. The majority of patients were HCV GT1 (68.7%), followed by HCV GT2 (30.4%). Ten different DAA regimens were used, with the majority receiving LDV/SOF (43.7%), followed by SOF+RBV (27.8%). One-third were treatment experienced (24.8% with prior PEG-IFN+RBV, 4.5% with prior DAA). SVR12 overall rate was 96.6%. Significant decreases noted in all major liver enzymes at week 12 and 24 post treatment. No increase in creatinine noted across treatments; 3.2% stopped treatment and 13.4% had an adverse event with fatigue (5.6% in patients treated with RBV vs. 6.4% in those treated without RBV, P=0.61) and anemia (5.6%) the most reported. Table 2 displays SVR12 rates by cirrhosis and prior treatment status for the most commonly used DAA treatments for GT1 and GT2 patients. SVR12 rates were excellent ranging from 97.1% (95%CI: 94.1-98.8%) to 99.7% (95%CI: 99.0-99.9%) for GT1 patients treated with LDV/SOF who did not have cirrhosis regardless of prior treatment history and who were treatment-naive with cirrhosis but lower in the cirrhotic treatment-experienced group (92.2%; 95%CI: 86.7-95.9%) (P<0.0001). Sub-analysis results for GT1b were similar, with SVR12 99.7% for non-cirrhotic treatment-naive, 99.5% for non-cirrhotic treatment-experienced, 97.4% for cirrhotic treatment-naive, and 93.0% for cirrhotic treatment-experienced, (P<0.0001). For GT2 patients, SVR12 was excellent for all groups (96.8-98.0%) except for cirrhotic treatment-experienced patients (n=66) who experienced an SVR12 of 87.9% (95%CI: 77.5-94.6%) (P=0.002). Conclusions: HCV cure rates were high overall in the REALC cohort-LDV/SOF GT1 98%; SOF+RBV GT2 96% except for cirrhotic, treatment-experienced patients especially in GT2, suggesting alternative therapy is needed.