SEN virus infection in patients with hepatocellular carcinoma

Although most cases of hepatocellular carcinoma (HCC) are associated with either the hepatitis B or C viruses (HBV, HCV), about 10-20% of HCCs occur in patients with chronic hepatitis that is aetiologically undefined. The aim of the present study was to determine the prevalence of the transfusion-transmitted SEN virus (SEN-V) in patients with HCC, including those patients who do not otherwise appear to be infected with HBV or HCV. Fragments of SEN-V subtypes D and H were amplified separately by PCR from the sera of 50 patients with HCC (31 from Canada and 19 from Japan) as well as from HCC and adjacent nontumourous liver tissues from eight of the Canadian patients. SEN-V DNA was found in the serum of 10 of 31 (32%) Canadian patients and eight of 19 (42%) Japanese patients [overall, 18 of 50 (36%) HCC patients]. SEN-V DNA was detected in the serum of 10 of 23 (43%) HCC patients with antibody to HCV (anti-HCV), six of 11 (55%) with hepatitis B surface antigen (HBsAg), and two of 16 (12%) without detectable anti-HCV or HBsAg. Twenty-three HCC patients in this study had 'silent HBV,' characterized by the detection of HBV DNA in the absence of HBsAg; eight of these (35%) also had SEN-V infections. SEN-V DNA was detected in HCC patients most typically in those with coexistent HBV or HCV infection. SEN-V was found in only one of seven HCC patients without HBV (without HBsAg or HBV DNA) or HCV and thus does not appear to be an important cause of 'cryptogenic' HCC.     

SEN virus infection in patients with chronic liver disease and hepatocellular carcinoma in Thailand

Background: SEN virus (SENV) has been recently identified as a candidate agent of non-A-E hepatitis virus. However, the exact role of this novel virus in the pathogenesis of chronic liver disease, including chronic hepatitis and cirrhosis, and the development of hepatocellular carcinoma (HCC) remains to be established. Methods: Using seminested polymerase chain reaction (PCR) amplification to detect SENV-D and SENV-H strains in serum, we investigated SENV infection in voluntary blood donors and in patients with chronic liver disease and HCC. Results: SENV was detected in 5 of 100 blood donors (5%), in 15 of 60 patients with chronic liver disease (25%), and in 25 of 60 patients with HCC (42%). The prevalence of SENV in patients with HCC was higher than that in patients with chronic liver disease (P = 0.05) and in blood donors (P < 0.001). An age-specific prevalence of SENV was found at high levels among individuals aged 21–40 years, but was not detected among individuals in the lower age group. No differences between SENV-infected and non-infected patients were demonstrated with respect to demographic data, assumed source of infection, biochemical abnormalities, and severity of chronic liver disease and HCC. Moreover, SENV infection had no apparent effect on the survival of patients with HCC. Conclusions: Our data suggest that SENV infection is frequent among patients with chronic liver disease and HCC. However, pathogenic effects associated with SENV infection in chronic liver disease and HCC need further investigation. Received: April 4, 2002 / Accepted: July 26, 2002 Acknowledgments. We would like to express our gratitude to the entire staff of the Viral Hepatitis Research Unit, Department of Pediatrics, Faculty of Medicine, Chulalongkorn University, for their tireless effort in this research project. We also would like to thank the Center of Excellence Fund, Rachadaphisek Somphot Endowment, Chulalongkorn University, and the Thailand Research Fund, Senior Research Scholar, for supporting this research. Reprint requests to: Y. Poovorawan     

TT virus infection among Egyptian patients with hepatocellular carcinoma

Hepatitis B and C viruses (HBV and HCV) have been associated with hepatocellular carcinoma (HCC). Recently, a novel DNA virus was isolated from a patient with posttransfusion hepatitis of unknown etiology and designated TT virus (TTV). To examine whether this virus is associated with HCC, we investigated sera from 82 Egyptian patients with histopathologically-diagnosed HCC. All subjects underwent serological investigations for detection of hepatitis B surface antigen (HbsAg), hepatitis B core antibody (HbcAb) and anti-HCV. Detection of TTV-DNA was performed by semi-nested polymerase chain reaction (PCR) using TTV-specific primers. TTV-DNA was detected in 28% of the patients. Age, gender, risk factors and biochemical liver functions did not significantly differ between TTV-DNA positive and negative patients. TTV was detected in 27.1% of patients with HCV-HCC, 25% of HBV-HCC, 66.7% of dual HCV and HBV infection and 40% of those with non-B, non-C-HCC (NBNC-HCC). It is concluded that, in this the cohort of Egyptian patients with HCC, TTV infection is common and is not associated with HCV, HBV, NBNC-HCC, history of schistosomiasis or blood transfusion.     

SEN virus infection and the risk of hepatocellular carcinoma: a case-control study

OBJECTIVE: Hepatitis B virus (HBV) and hepatitis C virus (HCV) are major risk factors for hepatocellular carcinoma (HCC). The role of a novel DNA virus, designated SEN virus (SENV), in the etiology of liver cancer remains to be established. The aim of this study was to evaluate the association between SENV infection and the risk of HCC by conducting a hospital-based, case-control study among Thai patients. METHODS: Eighty-six patients with HCC were enrolled and matched individually to a control according to sex, age (+/- 5 yr), and geographic background. The presences of HBV DNA, HCV RNA, and SENV DNA in stored serum samples were detected with the use of semi-nested polymerase chain reaction amplification. RESULTS: Individuals who were infected with SENV did not have increased risk of developing HCC (OR=1.49, 95% CI=0.50-4.42). In contrast, those who were positive for HBV markers (hepatitis B surface antigen and/or HBV DNA) or HCV markers (anti-HCV and/or HCV RNA) had significant risk for HCC (OR=19.91, 95% CI=8.26-47.98 and OR=7.97, 95% CI=2.15-29.54, respectively). Moreover, coinfection with SENV did not further increase the risk of HCC among patients infected with HBV and/or HCV. CONCLUSION: Our data suggest that, unlike chronic HBV or HCV infection, SENV infection is not a risk factor for developing HCC in Thai populations.     

Determinants of survival following hepatocellular carcinoma in Egyptian patients with untreated chronic HCV infection in the pre-DAA era

Background and Study Aims

In this study we assessed rates and determinants of survival in people with untreated chronic HCV infection and hepatocellular carcinoma (HCC) in an Egyptian liver clinic setting.

Caspase recruitment domains. New potential markers for diagnosis of hepatocellular carcinoma associated with HCV in Egyptian patients

Background and rational for the study. Chronic HCV is a major cause of HCC development. Caspase Recruitment Domains (CARD) is protein modules that regulate apoptosis and play an important role in various carcinogenesis processes, our aim is to assess the possible role of CARD9, CARD10 and Caspase only protein (COP) in progression of liver fibrosis and pathogenesis of HCC in Egyptian chronic HCV patients.Material and methods. 130 patients were recruited and classified into 4 groups; I: chronic HCV, II: chronic active hepatitis, III: liver cirrhosis, IV: HCV related HCC. Biochemical, virological studies, abdominal ultrasonography and liver biopsy were performed. Quantitative estimation of mRNA of CARD9, CARD10 and COP gene expression was performed by RT- PCR in liver biopsy from all patients.Results. In HCC patients; age, AFP and liver profile were significantly higher, HB and platelets were significantly lower (p value <0.01). The expression levels of mRNA of CARD9, CARD10 and COP in liver biopsies of HCC were significantly higher than other groups with direct correlation with age and no correlation with AFP, viral load, liver fibrosis or necroinflammatory activity. On differentiation between HCC and non HCC patients each CARD was assessed separately and combined, on combing the 3 CARDs, the sensitivity was 100%, specificity was 48%, positive predictive value 47% and negative predictive value 100%.Conclusions. CARD9, CARD10 and COP had no role in liver fibrosis but may be involved in hepatic carcinogenesis and they could be used as markers for HCC diagnosis and candid genes for molecular target therapy.     

SEN virus does not affect treatment response in hepatitis C virus coinfected patients but SEN virus response depends on SEN virus DNA concentration

AIM: To clarify the effect of SEN virus (SENV) infection on a combination therapy including interferon alfa (IFN-α) or pegylated-IFN with ribavirin in patients with chronic hepatitis and the effect of a combination therapy on SENV.METHODS: SENV DNA was determined by polymerase chain reaction in serum samples from 95 patients with chronic hepatitis C. Quantitative analysis was done for SENV H DNA.RESULTS: Twenty-one (22%) of 95 patients were positive for SENV DNA. There was no difference in clinical and biochemical parameters between patients with HCV infection alone and coinfected patients. The sustained response rate for HCV clearance after combination therapy did not differ between patients with SENV (52%) and without SENV(50%, n.s.). SENV DNA was undetectable in 76% of the initially SENV positive patients at the end of follow-up. SENV H response to combination therapy was significantly correlated with SENV DNA level (P=-0.05).CONCLUSION: SENV infection had no influence on the HCV sustained response rate to the combination therapy.Response rate of SENV to the combination therapy depends on SENV DNA level.     

Effect of SEN virus coinfection on outcome of lamivudine therapy in patients with hepatitis B

AIM: Interactions between hepatitis B virus (HBV) and other viral hepatitis infections are well known, whether the newly discovered SEN virus (SENV) has any effect on lamivudine antiHBV activity is unclear. Our aim was to clarify the effect on treatment outcome of coinfection with SEN virus in patients with hepatitis B during lamivudine therapy. METHODS: Nested polymerase chain reaction (PCR) amplification was used to detect SENV-D and SENV-H strains in serum from 45 patients with chronic hepatitis B treated with lamivudine 100 mg daily for 12 mo. HBV DNA load was detected with fluorescence quantitative PCR (FQ-PCR) and YMDD (tyrosine, methionine, aspartate, aspartate) motif mutation of HBV DNA was investigated with cDNA microarray. RESULTS: SENV DNA was detected in 5 of 45(11.1%) cases after 12 mo they received lamivudine treatment. SENV-D and SENV-H were 4.4% and 6.7% respectively. HBV DNA failed to respond to lamivudine therapy in 4 of 5 SENV coinfected patients while only 10 of 40 patients became SENV positive and the difference was statistically significant. Response of ALT and HBeAg to lamivudine had no significant difference between coinfection patients and single HBV infection ones. CONCLUSION: Coinfection with SEN virus in chronic hepatitis B patients may adversely affect the outcome of lamivudine treatment.     

Hepatitis C virus (HCV) genotypes in Spanish patients with HCV infection: relationship between HCV genotype 1b, cirrhosis and hepatocellular carcinoma

Background/Aims: The influence of the infecting virus genotype on the progression of the underlying liver disease in patients with chronic hepatitis C virus (HCV) infection remains controversial. The aim of this study was to investigate the prevalence of HCV genotypes in Spanish patients with chronic HCV infection and to elucidate the relationship between the infecting genotype and severity of the disease.Methods: A cross-sectional, retrospective analysis of frequency distribution of HCV genotypes was carried out in 414 Spanish patients with chronic HCV infection, including 243 patients with asymptomatic or minimally symptomatic chronic hepatitis, 112 patients with cirrhosis and hepatocellular carcinoma and 59 patients with decompensated cirrhosis. HCV genotype was determined by restriction fragment length polymorphisms of the 5′ non-coding region.Results: Infection with HCV genotype 1b was found in 72% of patients with chronic hepatitis and in more than 90% of patients with cirrhosis, with or without hepatocellular carcinoma. Older age, infection with genotype 1b and absence of overt parenteral exposure as a possible source of infection were associated with cirrhosis and hepatocellular carcinoma by univariate analysis and this association was confirmed by regression analysis.Conclusions: HCV genotype 1b is associated with advanced liver disease in our geographical area. However, this may be related to a cohort-effect caused by over-representation of genotype 1b in older patients with more advanced disease, because, in our country, this HCV genotype appeared earlier in time and is therefore associated with more prolonged periods of infection.     

Natural and activated cytotoxic lymphocytes reactivity to human hepatocellular carcinoma cell lines in hepatocellular carcinoma patients

The status of cellular cytotoxic activity in Hepatocellular Carcinoma (HCC) patients was compared to that in normal individuals by testing the cytotoxicity against K562 and five established HCC cell line targets. Natural killer (NK) activity of fresh peripheral blood mononuclear (PBM) cells in HCC patients to K562 cell line target was lower than that in normal donors. NK activity of unstimulated PBM cells from either source was minute against all five HCC cell line targets. Three different activation systems were employed to examine the cellular cytotoxicity of activated PBM cells: (1) conventional mixed lymphocyte culture (MLC), (2) allogeneic mixed lymphocyte tumor culture (MLTC), and (3) lymphokine-activated killer (LAK) cell culture. The cytotoxic effects of PBM cells in all three activation conditions were significantly lower in HCC patients than in normal donors (P less than 0.05 to P less than 0.01). These results suggest that, in addition to naturally present NK cells, the degree of in vitro activation of PBM cells may also have decreased in HCC patients.     

Effect of surveillance for hepatocellular carcinoma on tumor staging and treatment decisions in Egyptian patients

Purpose  

Egyptian hepatocellular carcinoma (HCC) patients present at advanced stages. We aimed to study the influence of surveillance versus non-surveillance on HCC staging and the potential therapeutic options.     

Impact of radiation and hepatitis virus infection on risk of hepatocellular carcinoma

In cohort studies of atomic bomb survivors and Mayak nuclear facility workers, radiation-associated increases in liver cancer risk were observed, but hepatitis B virus (HBV) and hepatitis C virus (HCV) infections were not taken strictly into account. We identified 359 hepatocellular carcinoma (HCC) cases between 1970 and 2002 in the cohort of atomic bomb survivors and estimated cumulative incidence of HCC by radiation dose. To investigate contributions of radiation exposure and hepatitis virus infection to HCC risk, we conducted a nested case-control study using sera stored before HCC diagnosis in the longitudinal cohort of atomic bomb survivors. The study included 224 HCC cases and 644 controls that were matched to the cases on gender, age, city, and time and method of serum storage, and countermatched on radiation dose. The cumulative incidence of HCC by follow-up time and age increased significantly with radiation dose. The relative risk (RR) of HCC for radiation at 1 Gy was 1.67 (95% confidence interval: 1.22-2.35) with adjustment for alcohol consumption, body mass index (BMI), and smoking habit, whereas the RRs for HBV or HCV infection alone were 63 (20-241) and 83 (36-231) with such adjustment, respectively. Those estimates changed little when radiation and hepatitis virus infection were fit simultaneously. The RR of non-B, non-C HCC at 1 Gy was 1.90 (1.02-3.92) without adjustment for alcohol consumption, BMI, or smoking habit and 2.74 (1.26-7.04) with such adjustment. CONCLUSION: These results indicate that radiation exposure and HBV and HCV infection are associated independently with increased HCC risk. In particular, radiation exposure was a significant risk factor for non-B, non-C HCC with no apparent confounding by alcohol consumption, BMI, or smoking habit.     

Impact of antiviral therapy on post-hepatectomy outcome for hepatitis B-related hepatocellular carcinoma

The outcome after curative resection for hepatocellular carcinoma (HCC) remains unsatisfactory due to the high recurrence rate after surgery. In patients with hepatitis B virus (HBV)-related HCC, which is the majority of patients with HCC in Asia, a high viral load is a strong risk factor for HCC recurrence. It is logical to believe that antiviral therapy may improve the post-operative outcome by promoting viral clearance and hepatocyte regeneration, as well as improving residual liver volume in HCC patients with hepatitis B. However, the effect of antiviral therapy on clinical outcomes after liver resection in patients with HBV-related HCC remains to be established. There are two main groups of antiviral treatment for HBV-oral nucleos(t)ide analogues and interferon. Interferon treatment reduces the overall incidence of HBV-related HCC in sustained responders. However, side effects may limit its long-term clinical application. Nucleos(t)ide analogues carry fewer side effects and are potent in terms of viral suppression when compared to interferon and are typically implemented for patients with more advanced liver diseases. They may also improve the outcome after curative resection for HBV-related HCC. There are increasing evidence to suggest that antiviral therapy could suppress HBV, decrease the perioperative reactivation of viral replication, reduce liver injury, preserve the liver function before and after operation, and may lower the risk of HCC recurrence. After all, antiviral therapy may improve the survival after liver resection by reducing recurrence and delaying the liver damage by the virus, resulting in a higher chance of receiving aggressive salvage therapy during HCC recurrence.     

Annexin A2 as a biomarker for hepatocellular carcinoma in Egyptian patients

AIM To investigate the clinical utility of serum annexin A2(ANXA2) as a diagnostic marker for early hepatocellular carcinoma(HCC).METHODS This study was performed in HCC Clinic of Ain Shams University Hospitals, Cairo, Egypt and included: Group 1: Fifty patients with early stage HCC(Barcelona Clinic Liver Cancer stage A); Group 2: Twenty five patients with chronic liver disease; and Control Group: Fifteen healthy, age-and sex-matched subjects who were seronegative for viral hepatitis markers. The followinglaboratory investigations were done: Viral hepatitis markers [hepatitis B surface antigen and hepatitis C virus(HCV) antibodies], HCV RNA in HCV antibody-positive patients, serum alpha fetoprotein(AFP), and serum ANXA2 levels.RESULTS In this study, 88% of HCC patients(n = 44) were HCVpositive, while HBV infection represented only 8% of all HCC patients(n = 4); and two patients were negative for both viral markers. A highly significant difference was found between patients with HCC and chronic liver disease as well as controls with regard to serum ANXA2 levels(130, IQR 15-240; 15, IQR 15-17; and 17, IQR 15-30 ng/m L, respectively). The area under the curve of ANXA2 was 0.865; the cut-off value was established to be 18 ng/mL with a diagnostic sensitivity of 74% and a specificity of 88%, while the sensitivity and specificity of AFP at the cut-off value of 200 ng/dL were 20% and 100%, respectively.CONCLUSION Serum ANXA2 may serve as a biomarker for the early detection of HCC.     

Impact of obesity on the surgical outcome following repeat hepatic resection in Japanese patients with recurrent hepatocellular carcinoma

AIM： To evaluate the impact of obesity on the postoperative outcome after hepatic resection in patients with hepatocellular carcinoma （HCC）. METHODS： Data from 328 consecutive patients with primary HCC and 60 patients with recurrent HCC were studied. We compared the surgical outcomes between the non-obese group （body mass index： BMI 〈 25 kg/m2） and the obese group （BMI ≥ 25 kg/m2）. RESULTS： Following curative hepatectomy in patients with primary HCC, the incidence of postoperative complications and the long-term prognosis in the nonobese group （n = 240） were comparable to those in the obese group （n = 88）. Among patients with recurrent HCC, the incidence of postoperative complications after repeat hepatectomy was not significantly different between the non-obese group （n = 44） and the obese group （n = 16）. However, patients in the obese group showed a significantly poorer long-term prognosis than those in the non-obese group （P 〈 0.05, five-yearsurvival rate; 51.9% and 92.0%, respectively）. CONCLUSION： Obesity alone may not have an adverse effect on the surgical outcomes of patients with primary HCC. However, greater caution seems to be required when planning a repeat hepatectomy for obese patients with recurrent HCC.     

Impact of biomarkers on disease survival and progression in patients treated with octreotide for advanced hepatocellular carcinoma

Background Current determination of prognosis for advanced hepatocellular carcinoma (HCC) is mainly based on clinical assessment. We aimed to determine the impact of biomarkers as predictive factors for HCC progression and survival during octreotide-based treatments.Patients and methods We included patients who had been prospectively randomised to receive either octreotide (30 mg) alone monthly (n = 39) or in combination with rofecoxib (up to 50 mg bid daily, n = 32) for a minimum of 6 months, or until death occurred.Results Overall median survival (154 days) and median time to progression (94 days) were not different for both treatments and the biomarkers investigated (VEGF-A, IGF-1, PGE-2, ET-A) were similarly distributed amongst treatment groups. Combined univariate group analysis revealed that survival was decreased for an uptake ratio of > 2 on initial octreoscan (P = 0.05); baseline serum VEGF-A and IGF-1 were further significantly associated with survival. On multivariate analysis, uncorrected serum VEGF-A appeared to be the most significant predictor for tumor progression and survival.Conclusions Biomarkers, in addition to established tumor markers, are independent predictors of tumor progression and survival in patients with advanced HCC treated with octreotide. Furthermore, the involvement of VEGF-A implies the inhibition of angiogenesis as a potential mechanism of action for this drug.     

Association between HCV and HBV infection in hepatocellular carcinoma and alcoholic liver disease.

We conducted a retrospective study in 229 patients to determine the prevalence of hepatitis C virus (HCV) according to the type of liver disease and also the serum hepatitis B virus (HBV) status. There were 55 cases of hepatocellular carcinoma (HCC), 78 cases of alcoholic cirrhosis (AC) and 96 cases of non-cirrhotic alcohol liver disease (NCA). Half the AC and NCA groups were constituted of patients with or without serum HBV markers. The prevalence of anti-HCV antibodies in HCC (58.2%) was significantly higher than in the other groups (AC, 35.9%; NCA, 17.7%). Combining the two AC and NCA groups gave a 25.8% anti-HCV prevalence in the alcoholics. Altogether anti-HCV antibodies were more frequently present in HBV-positive than in HBV-negative patients (42.2 vs. 26.1%, p less than 0.01), although the difference was not significant when the HCC group was separately analysed. The S/N ratio of anti-HCV-positive samples varied according to the type of the disease (tumorous vs. non-tumorous) and to the serum HBV status. Indeed, 65.6% of HCC had a S/N greater than 4 compared to only 32.2 and 23.5% in AC and NCA, respectively (p less than 0.01). On the other hand more than half of the AC and NCA had a S/N less than 2, indicating possible artefacts in certain cases. The 15 HBsAg and anti-HCV-positive patients had a significantly lower mean S/N than patients displaying only antibodies against HBV, even in the HCC group.(ABSTRACT TRUNCATED AT 250 WORDS)     

Chronic liver disease and hepatitis C virus in Egyptian patients.

BACKGROUND/AIMS: This study was designed to evaluate anti-HCV and anti-GOR in chronic liver disease (CLD) caused by HCV alone or with bilharzia. METHODOLOGY: The parameters of hepatitis C virus (HCV) infection among 45 chronic liver disease (CLD) patients are the subject of this study. The samples that were collected included serum, saliva and liver biopsy. For comparison, 44 serum, saliva and liver biopsies were also collected from non liver disease (NLD) patients undergoing surgery at the Gastroenterology Surgical Center, Mansoura University. RESULTS: Screening of antihepatitis C (anti-HCV) with a second generation ELISA test showed that 37/45 (82.2%) sera and 17/45 (37.7%) saliva samples from CLD patients were positive for the presence of anti-HCV (IgG), while, anti-HCV (IgG) was detected among 32/44 (72.7%) sera and 6/44 (13.6%) saliva samples from NLD patients. HCV antigen was detected by immunostaining in the liver biopsy sections of 11/45 (24.4%) CLD and in 6/44 (13.6%) NLD patients. HCV antigen was detected in hepatocyte cytoplasm and nuclei, in some endothelial cells lining the hepatic cell cords, and in some bile duct cells. The serum and saliva samples from both CLD and NLD patients were also tested by ELISA for the presence of anti-GOR to determine the prevalence of autoantibody in HCV infected and non-infected patients. Anti-GOR was detected in 19/45 (42.2%) sera and in 1/45 (2.21%) saliva samples from CLD patients, while in the case of NLD patients, anti-GOR antibodies were found in 7/44 (15.9%) sera and in 4/44 (9%) saliva samples. GOR antigen was detected by an indirect immunoperoxidase stain of liver biopsies. Positive GOR antigen signals were found in hepatocytes but granular cytoplasmic, and extrahepatic localization was also noticed. A correlation between the detection of anti-GOR and anti-HCV revealed that, out of 37 anti-HCV positive CLD patients, there were 19 (51.3%) positive for anti-GOR, while 7/32 (21.8%) NLD patients were positive for anti-HCV and for anti-GOR. CONCLUSIONS: The results of the present study confirm the published anti-HCV high seropositivity among Egyptian CLD patients and point to an autoimmune processes in CLD. The liver biopsy findings stress the presence of HCV antigen in extra hepatic cells as well as in hepatocytes in CLD. Our data confirm that anti-GOR is commonly present in sera from CLD patients and show that anti-GOR are secreted in saliva. Our results showed that saliva can not be used reliably, instead of serum, for the diagnosis of HCV infection or auto-antibodies related to HCV infection, but can be used as a parameter for the evaluation of CLD activity, when repeated sampling is necessary.