Search

HBV nucleos(t)ide reverse transcriptase inhibitors (NrtIs) do not completely suppress HBV replication. Previous reports indicate persistent viremia during NrtI treatment despite HBV DNA being undetectable. HBV core inhibitors may enhance viral suppression when combined with NrtIs. This phase II trial (NCT03576066) evaluated the efficacy and safety of the investigational core inhibitor, vebicorvir (VBR), in virologically- suppressed patients on NrtIs.Non-cirrhotic, NrtI-suppressed patients with chronic HBV were randomised to VBR 300 mg once daily or matching placebo (PBO) for 24 weeks. Treatment was stratified by hepatitis B e antigen (HBeAg) status. The primary endpoint was change from Baseline in serum HBeAg or hepatitis B surface antigen (HBsAg) after 24 weeks.Of 73 patients enrolled, 47 were HBeAg positive and 26 were HBeAg negative. In HBeAg-positive and -negative patients, there were no differences in the change from Baseline at Week 24 for HBsAg or HBeAg. Using a novel, high-sensitivity assay to detect HBV DNA, a greater proportion of patients with detectable HBV DNA at Baseline achieved undetectable HBV DNA at Week 24 in the VBR+NrtI vs. PBO+NrtI group. In HBeAg-positive patients, a greater change from Baseline in HBV pregenomic (pg)RNA was observed at Week 24 with VBR+NrtI vs. PBO+NrtI. Treatment-emergent adverse events (TEAEs) in VBR+NrtI patients included upper respiratory tract infection, nausea, and pruritus. No serious adverse events, Grade 4 TEAEs, or deaths were reported.In this 24-week study, VBR+NrtI demonstrated a favourable safety and tolerability profile. While there were no significant changes in viral antigen levels, enhanced viral suppression was demonstrated by greater changes in DNA and pgRNA with the addition of VBR compared to NrtI alone.NCT03576066.Core inhibitors represent a novel approach for the treatment of chronic hepatitis B virus (HBV) infection, with mechanisms of action distinct from existing treatments. In this study, vebicorvir added to existing therapy reduced HBV replication to a greater extent than existing treatment and was generally safe and well tolerated.

Nucleos(t)ide reverse transcriptase inhibitors do not completely suppress HBV DNA in chronic HBV infection (cHBV). Vebicorvir (VBR) is an investigational core inhibitor that interferes with multiple aspects of HBV replication. This phase II trial evaluated the safety and efficacy of VBR in combination with entecavir (ETV) in treatment-naïve patients with cHBV.HBeAg-positive, treatment-naïve patients without cirrhosis were randomised 1:1 in a double-blind manner to once-daily VBR 300 mg+ETV 0.5 mg or placebo (PBO)+ETV 0.5 mg for 24 weeks. The primary endpoint was change in mean logAll patients in both treatment groups (PBO+ETV: 12/12; VBR+ETV: 13/13) completed the study. At Week 12, VBR+ETV led to a greater mean (SD) reduction from Baseline in logIn this 24-week study, VBR+ETV provided additive antiviral activity over PBO+ETV in treatment-naïve patients with cHBV, with a favourable safety and tolerability profile.NCT03577171 LAY SUMMARY: Hepatitis B is a long-lasting viral infection of the liver. Current treatments can suppress hepatitis B virus but do not offer the opportunity of cure, hence, new treatment approaches are required. Herein, we show that the combination of the novel core inhibitor vebicorvir with an existing antiviral (entecavir) in treatment-naïve patients chronically infected with hepatitis B virus demonstrated greater antiviral activity than entecavir alone. Additionally, vebicorvir was safe and well tolerated. Thus, further studies evaluating its potential role in the treatment of chronic hepatitis B are warranted.

Chronic hepatitis B virus (HBV) infection is a major public health challenge on the global scale. Affecting hundreds of millions worldwide, HBV is a leading risk factor for hepatocellular carcinoma (HCC). Clinical outcomes from chronic HBV infection are varied and appear to be influenced by a complex and dysregulated host immune response. In turn, much attention has been given to the immunologic response to HBV in an effort to identify host factors that lead to the development of HCC. However, the role of nonimmunologic host factors, such as chronic stress, in HBV-related HCC is poorly defined. Indeed, a growing appreciation for the effects of stress on chronic liver diseases raises the question of its role in chronic HBV infection. In this light, the present review will untangle the roles of key host factors in HBV-related HCC with an emphasis on chronic stress as a viable contributor. First discussed is the interplay of stress, inflammation and chronic liver disease. The host immune response’s role as a driver of HBV-related HCC is then reviewed, allowing for a close exploration of the effects of stress on immune function in chronic hepatitis B and as a potential risk factor for HBV-related HCC.

BACKGROUND: Cell-free DNA (cfDNA) from blood has become a promising analyte for cancer diagnostics. However, because blood contains high concentrations of proteins and enzyme inhibitors, plasma cfDNA quality can be variable. Urinary cfDNA has been shown to contain mutations associated with non-genitourologic cancers including hepatocellular carcinoma (HCC). In this study, we evaluate urine as a noninvasive alternative to blood-based liquid biopsy for both germline and ctDNA genotyping in HCC. METHOD: Using qPCR, whole-genome sequencing (WGS), and targeted NGS, DNA isolated from blood or urine of patients with HCC was evaluated and analyzed for overall genome coverage, HCC hotspot coverage, and germline or somatic mutation concordance. Targeted NGS of plasma and urine cfDNA was also performed for detection of somatic variants. RESULTS: Similar to plasma cfDNA, urine cfDNA showed a major mononucleosomal species of 165-170 bp, with dinucleosomal species also discernable in most healthy individuals and patients with HCC. Overall genome coverage breadth was similar between urine and plasma cfDNA, with higher fraction of covered cancer-associated mutation hotspots in urine cfDNA. Quantitative PCR analyses of HCC-associated mutations (TP53, CTNNB1, and hTERT) in 102 patients with HCC, revealed 78% overall concordance between plasma and urine. Targeted NGS of HCC-associated gene regions in additional 15 patients showed an 97% overall position-level concordance between plasma and urine cfDNA. Furthermore, urine outperformed plasma in both the sensitivity and specificity of detection of CTNNB1 and hTERT mutations. CONCLUSION urine DNA can potentially be used as a completely noninvasive diagnostic biomarker for HCC. Citation Format: Amy Kim, Selena Lin, Yixiao Cui, Terence Gade, Fwu-Shan Shieh, Max Chao, John Shieh, Surbhi Jain, Jonathan Cheng, James Hamilton, Hie-Won L. Hann, Dmitry Goryunov, Ying-Hsiu Su. Urine as a non-invasive alternative to blood for germline and somatic mutation detection in hepatocellular carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 544.

Background: Psychosocial stress is a common comorbidity in chronic hepatitis B (CHB) patients. A diagnosis of CHB, the primary risk factor for liver cancer among Asian Americans, can be a significant source of psychological and emotional stress. Although the disease can be life-changing, there is a paucity of research on psychosocial stress in CHB patients. The purpose of this study is to examine the prevalence of serious psychological distress (SPD) and factors associated with SPD in Asian CHB patients. Method: It is a prospective study design with subjects identified from an existing patient cohort that were enrolled and then subsequently followed up. Information was gathered from the CHB patients using face-to-face interviews in Korean or English. Serious psychological distress was measured by the Kessler Scale (K6). Measures of stressful life events and Hepatitis B Quality of Life (HBQOL) (e.g., psychological well-being, anticipation anxiety, stigma) were used. A multivariate logistic regression model (including gender, age, level of education) was conducted for analysis. Results. Of 50 CHB patients 18 years and older, the 12-month prevalence of SPD was estimated at 20% using the optimal cut-off (>=13). In adjusted regression analysis, stressful life events and HBQOL-stigma were related to serous psychological distress: Those who had stressful life events (aOR=3.10, 95% CI 1.26-7.64) and those who had high scores of stigma (aOR=1.19, 95% CI 0.97-1.44, p=.08) had higher psychological distress. Those with more than a college education had decreased psychological distress (aOR=0.09, 95% CI 0.01-0.82). Conclusion: The finding indicates CHB patients had a higher prevalence of serious psychological distress than the general Asian population (2.6% from California Health Interview Survey). Stigma connected to hepatitis B diagnosis and stressful life events were important factors associated with mental health. This suggests that disease burden caused by hepatitis B infection may lead to poor mental health. Future studies will identify those who are at a high likelihood of developing a mental disorder and develop effective interventions for early detection of mental illness. Citation Format: Hee-Soon Juon, Grace Park, Klassen Ann, Hie-Won Hann. Factors associated with psychological distress among chronic hepatitis B patients [abstract]. In: Proceedings of the Eleventh AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2018 Nov 2-5; New Orleans, LA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2020;29(6 Suppl):Abstract nr A102.

Objectives: It is known that chronic hepatitis B (CHB) infection exerts suppressive effects on the host innate and adaptive immune responses. Although there is information about liver disease progression of CHB patients and the adverse effects of stress on the immune system, few studies focus on the effects of chronic stress in CHB patients. To our best knowledge, this is the first study to measure hair cortisol in CHB patients. To address the prolonged exposure to stress associated with CHB diagnosis, we explored the measure of the stress hormone cortisol (CORT) in the hair as a biomarker of chronic stress in CHB patients. Methods: Eligible CHB patients identified from an existing patient cohort were enrolled in this prospective study. Data collection were done using medical chart reviews, face-to-face interview in Korean or English, and hair and blood samples. Hair samples were cut from the posterior vertex of the scalp, as close to the scalp as possible. Hair CORT in the most proximal 3-cm scalp hair segment was analyzed using luminescence immunoassay. Results: We collected hair samples of 50 CHB patients (age 34-74 years) with duration of HBV diagnosis (1-51 years, 23.8±10). Hair samples weighing 19.6 - 29.9 mg were processed and analyzed for cortisol. Mean cortisol levels were 16.42 pg/mg (27.59 SD) ranging from 2.7 to 168.1 pg/mg. About 16% had below normal HCC (=4) was 48%. Distress was positively correlated with hair CORT (r=.395, p Conclusion: About one fifth of the patients had very low hair cortisol levels, suggesting long-term suppression of hair cortisol production in the course of HBV diagnosis and antiviral therapy. Low hair cortisol level might indicate a specific pathogenic mechanism with cortisol production. This study suggests further research to determine the longitudinal effects of chronic stress on hair cortisol levels and chronic liver disease progression in CHB patients. Citation Format: Hie-Won Hann, Hushan Yang, Jerrold Meyer, Hee-Soon Juon. Hair cortisol measurement in chronic hepatitis B patients [abstract]. In: Proceedings of the Eleventh AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2018 Nov 2-5; New Orleans, LA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2020;29(6 Suppl):Abstract nr A101.

Since 1998 with the highly effective nucleos(t)ide analogues for Hepatitis B virus (HBV), we have witnessed successful viral suppression that has led to the delay or prevention of the development of hepatocellular carcinoma (HCC). Reduced incidence of HCC with anti-HBV treatment has been documented with lamivudine, entecavir and tenofovir. A number of investigators have observed that after the initial tumor was ablated, anti-HBV treatment could prevent or reduce the recurrent or subsequent new HCC. Nonetheless, as reported earlier, we are observing the persistent risk for HCC in patients in spite of successful viral suppression for over a decade with anti-HBV therapy. In this report, we present three cases who underwent initial tumor ablation and maintained negative HBV DNA with anti-HBV therapy and yet developed subsequent new or recurrent HCC at the intervals of 5,10,11 and 15 years. These persistent risks for new and recurrent HCC are attributed to the incomplete control of HBV due to the presence of cccDNA in the host’s hepatocytes. Current antiviral treatment can achieve a functional cure (suppression of HBV replication) while a complete cure of eliminating the cccDNA has not been possible. Therefore, even though HBV is not actively replicating, cccDNA remains in the nucleus of the hepatocytes and continues hepatocarcinogenic processes including HBV and host DNA integration. There is a need for HBV cure drugs.

// Chun Wang 1, 2, * , Hie-Won Hann 3, * , Zhong Ye 1, * , Richard S. Hann 3 , Shaogui Wan 1, 4 , Xishan Ye 1 , Peter D. Block 1 , Bingshan Li 5 , Ronald E. Myers 1 , Xiaowei Wang 6 , Hee-Soon Juon 1 , Jesse Civan 3 , Mimi Chang 7 , Ho S. Bae 7 , Jinliang Xing 8 , Hushan Yang 1 1 Division of Population Science, Department of Medical Oncology, Sidney Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, PA 19107, USA 2 Department of Environmental Health, School of Public Health, Nantong University, Nantong, Jiangsu 226000, China 3 Liver Disease Prevention Center, Division of Gastroenterology and Hepatology, Department of Medicine, Thomas Jefferson University, Philadelphia, PA 19107, USA 4 Institute of Pharmacy, Pharmaceutical College, Henan University, Kaifeng, Henan 475004, China 5 Center for Human Genetics Research, Department of Molecular Physiology & Biophysics, Vanderbilt University, Nashville, TN 37232, USA 6 Department of Radiation Oncology, Washington University School of Medicine, Saint Louis, MO 63108, USA 7 Asian Pacific Liver Center, Saint Vincent Medical Center, Los Angeles, CA 90057, USA 8 Experimental Teaching Center, School of Basic Medicine, Fourth Military Medical University, Xi’an 710032, China * These authors have contributed equally to this work Correspondence to: Hushan Yang, e-mail: hushan.yang@jefferson.edu Keywords: hepatocellular carcinoma, risk, microRNA, serum Received: December 15, 2015 Accepted: May 4, 2016 Published: May 24, 2016 ABSTRACT The predictive value of circulating microRNAs (miRNAs) in hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC) has been demonstrated in retrospective studies, but it has rarely been tested in prospective studies. In a cohort of 373 cancer-free HBV patients with a median follow-up of 4.5 years, we measured the expression of 24 retrospectively identified HCC-related miRNAs in baseline serum samples. When we analyzed the prospective associations of miRNA expression with HCC risk using the Cox proportional hazards model, we found that 15 of the 24 miRNAs exhibited a significant association with HCC risk. In particular, 7 miRNAs (miR-122, miR-99a, miR-331, miR-125b, miR-23b, miR-92a, and miR-26a) were associated with an increased risk, and 8 miRNAs (miR-652, miR-23a, miR-27a, miR-34a, miR-145, miR-10a, miR-150, and let-7f) were associated with a decreased risk. Compared to HBV patients with a low miRNA-based risk score, those with a high miRNA-based risk score exhibited a significantly elevated HCC risk in both univariate (hazard ratio [HR] 6.56, 95% confidence interval [CI] 2.74-15.70) and multivariate (HR 3.57, 95% CI 1.34-9.48) analyses. The risk score significantly increased the HCC prediction performance of alpha-fetoprotein (concordance index increased from 0.68 to 0.82, P < 0.0001). In silico analyses indicated that the genes targeted by the 15 miRNAs are mainly enriched in the transforming growth factor-beta signaling pathway. Collectively, these results provide prospective evidence that circulating miRNAs serve as non-invasive markers for risk prediction of HCC in HBV patients.

Simple hepatic cysts occur in less than 5% of the population. As the technology we use for imaging advances, so does the frequency with which we identify them. Simple cysts, as presented here, do not generally require aspiration. The majority of these cysts do not require treatment, however, those larger than 4 cm are recommended for surveillance. We present two cases of simple cysts that were followed for 15 years and managed differently. The experience we acquired from these cases illustrates challenges in understanding and management and an implication that, perhaps, conservative surveillance might be best.

To retrospectively evaluate the effect of indeterminate or false-negative findings at magnetic resonance (MR) imaging on eligibility for curative treatment of hepatocellular carcinoma (HCC).This HIPAA-compliant retrospective study was approved by the institutional review board; the need for informed consent was waived. Of 166 patients with cirrhosis in whom HCC was detected with MR imaging, 21 (13 men, eight women; mean age, 60 years) had 33 proved HCCs that were not detected on previous MR images obtained 6-24 months earlier. MR imaging included T1-weighted, T2-weighted, and dynamic contrast material-enhanced T1-weighted imaging. Serial MR images and treatment records were reviewed to evaluate nodule growth and the effect of delayed diagnosis on treatment eligibility.Of 33 HCCs in 21 patients, 24 corresponding nodules (73%) were described on previous MR images as benign or indeterminate. Five additional nodules were visible at retrospective evaluation, but only on arterial phase images. The diameters of these 29 visible but indeterminate nodules were initially 0.6-1.9 cm (mean, 1.1 cm) and increased to 0.9-4.5 cm (mean, 1.9 cm) at HCC diagnosis (mean follow-up, 378 days). The mean doubling time was 856 days for diameter and 285 days for volume. All nine HCCs with a delayed diagnosis of less than 1 year were smaller than 3 cm at diagnosis, and the patients had undergone liver transplantation (n=3) or technically successful ablation or embolization (n=6). All 10 subcentimeter indeterminate nodules were smaller than 2 cm at HCC diagnosis, and none progressed to untreatable HCC.Indeterminate nodules smaller than 2 cm did not become untreatable HCC with delayed HCC diagnosis of 6-12 months.

SUMMARY The Ag system, inherited antigenic determinants on serum low density lipoprotein and Australia antigen (Au), an antigen associated with an infectious agent which can cause hepatitis in man were discovered by reacting in immunodiffusion tests sera from multiply transfused patients and sera from geographically human populations. The approach has been pursued in the experiments described herein and has resulted in the identification of seventeen precipitating systems. They are different from the previously described anti-Au and anti-Ag precipitins. In some cases they may be confused with Australia antigen precipitins, and care should be taken in the evaluation of Au test systems where comparisons with antigen controls are not routinely performed.

Background and Aims: Chronic hepatitis B (CHB) is an important cause of end stage liver disease and hepatocellular carcinoma. Controlled clinical trials indicate treatment with lamivudine results in positive clinical responses. The study goal was to determine if the response to lamivudine treatment (HBeAg loss, HBV DNA loss and alanine aminotransferase [ALT] reduction) differs according to pretherapy (pre-tx) ALT levels. Methods: This was a retrospective review of medical record data. All CHB patients at all stages of disease (including cirrhotic) with more than two visits to the clinic were included in the study (n = 719). Kaplan-Meier survival and Cox proportional hazards were estimated. Results: Of the total 719 HBsAg (+) patients, 317 were treated with lamivudine 150 mg or 100 mg daily. Among HBeAg positive patients, at 3 years, Kaplan-Meier estimates of the loss of HBeAg were 40%, 57% and 61% for pre-tx ALT 2 × ULN, respectively. Similar results of HBV-DNA loss were seen in HBeAg negative patients. Conclusions: In this setting, we observed that pre-tx ALT levels were not associated with response to lamivudine, but that lower platelet count and female sex in HBeAg (+) patients were important predictive factors of a favorable response to lamivudine therapy.

Background and aims: Early detection of Hepatocellular Carcinoma (HCC) is crucial for effective management. Incidence of HCC has increased in the United States largely attributed to hepatitis B and C virus. Lens culinaris agglutinin-reactive Alpha-Fetoprotein (AFP-L3) and Des-Gamma-Carboxy Prothrombin (DCP) are being recognized specific biomarkers for HCC. Methods: We measured AFP-L3 and DCP in serial serum specimens of a cohort of chronic hepatitis patients on HCC surveillance and compared these markers to abdominal imaging. Among fifty patients who developed HCC during surveillance, 30 were included in the study with available sera 1-2 years before, at diagnosis and post ablation of HCC. For controls, three consecutive annual sera were examined from 106 chronic hepatitis patients without HCC during surveillance for 5-10 years. The μTASWako i30 auto analyzer was used for the assay that utilizes the microfluidics chip based assay platform. It can fractionate AFP-L3 glycoform and calculates AFP-L3% if AFP level is ≥ 0.6 ng/mL. Results: Combination of AFP, AFP-L3 and DCP showed high sensitivity of 83% in all patients and 75% in patients with AFP

Although the nucleoside analogue lamivudine has shown promise in patients with chronic hepatitis B, long-term data on patients from the United States are lacking. We randomly assigned previously untreated patients with chronic hepatitis B to receive either 100 mg of oral lamivudine or placebo daily for 52 weeks. We then followed them for an additional 16 weeks to evaluate post-treatment safety and the durability of responses. The primary end point with respect to efficacy was a reduction of at least 2 points in the score on the Histologic Activity Index. On this scale, scores can range from 0 (normal) to 22 (most severe abnormalities).Of the 143 randomized patients, 137 were included in the efficacy analysis: 66 in the lamivudine group and 71 in the placebo group. The other six patients were excluded at the base-line visit because of the absence of a documented history of hepatitis B surface antigen for at least six months. After 52 weeks of treatment, lamivudine recipients were more likely than placebo recipients to have a histologic response (52 percent vs. 23 percent, P0.001), loss of hepatitis B e antigen (HBeAg) in serum (32 percent vs. 11 percent, P=0.003), sustained suppression of serum hepatitis B virus (HBV) DNA to undetectable levels (44 percent vs. 16 percent, P0.001), and sustained normalization of serum alanine aminotransferase levels (41 percent vs. 7 percent, P0.001), and they were less likely to have increased hepatic fibrosis (5 percent vs. 20 percent, P=0.01). Lamivudine recipients were also more likely to undergo HBeAg seroconversion, defined as the loss of HBeAg, undetectable levels of serum HBV DNA, and the appearance of antibodies against HBeAg (17 percent vs. 6 percent, P=0.04). HBeAg responses persisted in most patients for 16 weeks after the discontinuation of treatment. Lamivudine was well tolerated. Self-limited post-treatment elevations in serum alanine aminotransferase were more common in lamivudine recipients: 25 percent had serum alanine aminotransferase levels that were at least three times base-line levels, as compared with 8 percent of placebo recipients (P=0.01). The clinical condition of all patients remained stable during the study.In U.S. patients with previously untreated chronic hepatitis B, one year of lamivudine therapy had favorable effects on histologic, virologic, and biochemical features of the disease and was well tolerated. HBeAg responses were generally sustained after treatment.

To determine whether the frequency of hepatocellular carcinoma (HCC) in patients with cirrhosis is affected by hepatic iron deposition as detected with magnetic resonance (MR) imaging.In a retrospective search of MR imaging and histopathology records, 196 patients with histopathologically proved cirrhosis and with (n = 80) or without (n = 116) HCC who underwent T2-weighted conventional or fast spin-echo and gradient-echo (GRE) (echo timeor = 6.0 msec) imaging were identified. MR images were qualitatively and quantitatively evaluated for diffuse hepatic iron deposition and siderotic regenerative nodules to assess their correlation with the presence of HCC.Hepatic parenchymal iron deposition was seen in 79 (40%) patients, and iron deposition in regenerative nodules was seen in 71 (36%) at MR imaging. The mean signal intensity ratio of GRE images in patients with hepatic iron deposition was significantly lower than that in patients without it (P.001). The frequency of HCC in patients with iron deposition in regenerative nodules (52% [37 of 71 patients]) was significantly higher (P = .015) than that in patients without iron in regenerative nodules (34% [43 of 125 patients]).The occurrence of HCC may be associated causally with iron deposition in regenerative nodules in patients with cirrhosis. MR imaging can enable detection of iron deposition in regenerative nodules as a possible risk factor for the development of HCC.

Hepataocellular carcinoma (HCC) is one of the top five common cancers among Asian Americans. The majority of the HCCs in this population are associated causally with chronic hepatitis B virus (HBV) infection. With the newly available antiviral agents, by eliminating the virus at an early stage of HBV infection, one could attempt to delay or prevent the development of HCC. The primary prevention of HCC would be a hepatitis B vaccine.

Chronic hepatitis B virus infection is endemicin Asian communities in the United States. The purposeof the current study was to compare the antiviralefficacy of interferon-α2b in a groupof adult Asian patients chronically infected withhepatitis B with active replication compared to acontrol group of Caucasian patients treated with thesame regimen. Patients with entry aminotransferase (ALT)levels greater than three times the upper limit ofnormal received interferon-α2b, 5million units, subcutaneously daily for 16 weeks.Patients with pretreatment ALT levels 1.5-3 times theupper limit of normal received prednisone for a total of sixweeks prior to interferon starting at 60 mg daily withreduction in dosage by 20 mg every two weeks with atwo-week period between finishing prednisone and starting interferon-α2b. Eight(62%) of the 13 Asians and six (60%) of the 10Caucasians cleared HBeAg and HBV DNA from serum (NS). Bythe end of one year of follow-up after therapy, four(67%) of six Caucasian responders but none of the Asianresponders had cleared hepatitis B surface antigen fromserum (P < 0.05). Loss of serum markers of activereplication appeared less durable in the Asian responders compared to the Caucasians withreappearance of serum HBeAg in two (25%) of eight of theformer but only one (17%) of the latter group. Threeother Asian patients subsequently redeveloped HBeAg in serum. It is concluded that adultAsian-Americans have an identical initial response rateto antiviral therapy withinterferon-α2b; however, the responsemay be less durable and does not usually lead to loss of HBsAg.

Earlier clinical studies have reported an ALT flare greater than 10 times the upper limit of normal in some patients with chronic hepatitis B when their lamivudine (LAM) treatment was switched to adefovir (ADV) therapy. The current study compared the safety of switching directly to ADV versus overlapping LAM and ADV for 3 months followed by ADV monotherapy. Patients with chronic hepatitis B receiving LAM therapy for ≥6 months were eligible for the study regardless of the presence of LAM resistance, HBeAg status or serum ALT levels. Eighteen patients (13 males) were randomized to direct switch to ADV and 17 patients (10 males) to overlap. HBV-DNA, ALT, albumin, and total bilirubin were assayed at baseline, 3, 6, 9, and 12 months. Study drugs were discontinued at the end of 12 months with the follow up at 3 and 6 months. The decision to continue antiviral therapy was made at the discretion of the investigator. Baseline ALT levels were similar between the direct switch and overlap group: median ALT (U/L) was 44.0 (16–266) and 33.0 (19–367) for direct switch for overlap group, respectively (P = 0.42). No ALT flare was noted at 3 months in either group: median ALT decreased from 44.0 to 34.5 U/L in the direct switch group, and from 33.0 to 23.0 in the overlap group. Furthermore, no patient in either group exhibited ALT flare throughout the 12 months. This study did not show an ALT flare during switch to ADV at 3 months or at any time later. J. Med. Virol. 82:1489–1493, 2010. © 2010 Wiley-Liss, Inc.

BACKGROUND Iron is essential for the growth of all living cells. One of the important intracellular roles for iron is in the activation of ribonucleotide reductase, the enzyme that catalyzes the first step in DNA synthesis. Thus, the intracellular iron level may serve as a regulator of cell growth. The authors tested the hypothesis that lowering body iron concentration inhibits the growth of human-derived hepatocellular carcinoma (HCC) cells by depleting these cells of iron. Deferoxamine (DFO), an iron-chelating agent, was used to lower intracellular iron level. METHODS HCC cells, PLC/PRF/5 (7 x 10(6) cells/mouse), were transplanted subcutaneously into athymic nude mice. When tumors reached 200-300 microliters in size, mice with comparable tumor sizes were paired; one was treated with DFO (300 mg/kg body weight/day, 5 days/week) intraperitoneally while the other received no treatment. RESULTS Eight pairs of mice with HCC were observed for 5-18 weeks. Mean tumor growth rates (TGR) (mean +/- standard error) for the untreated and treated mice were 30.5 +/- 3.7 microliters/week and 11.9 +/- 1.5 microliters/week. The difference was significant (P < 0.02). In the second set of studies, DFO treatment was begun when the tumor size was smaller (100-200 microliters). Four pairs of mice were observed for 4-15 weeks; mean TGR for the four untreated mice was 18.1 +/- 5.1 microliters/week. In two mice treated with DFO, tumors regressed completely by the seventh week after initiation of treatment. The two remaining mice on DFO therapy had much slower growing tumors, with a mean TGR of 1.8 +/- 0.5 microliters/week. CONCLUSIONS Thus, our results suggest that (1) reduction of intracellular iron concentration by DFO may be useful as antitumor therapy in HCC and (2) the favorable effects of DFO treatment are best seen when treatment is begun when the tumor is small.

Iron is essential for the growth of all cells, including tumor cells. The authors previously reported that a variety of transplantable tumors grew faster and larger in mice that were on an iron-rich diet compared with those on an iron-deficient diet. In this study the authors examined the relationship between iron in the diet and development of tumors in mice that are known to develop spontaneous tumors--C3H/HeN-MTV+(C3H-MTV+) mice that were congenitally infected with mammary tumor virus. These mice have a greater than 96% chance of developing mammary tumors between the ages of 7.2 and 9.2 months. Fifteen C3H-MTV+ weanlings were given a low-iron diet (5 mg iron/kg diet), and 15 were given diets with normal amounts of iron (180 mg Fe/kg diet). Thirteen of the 15 mice from the low-iron group and all 15 mice from the normal-iron group developed tumors. The average tumor growth rate in the normal-iron group was 112%/wk, compared with 62%/wk for the low-iron group. The difference in tumor growth rate between the two groups was significant (P = 0.02 by Student's t test). In this study, low iron intake did not prevent tumor development, but the results confirm the authors' previous report that iron nutrition of the host affects tumor growth; tumors grow better in an iron-rich environment. High levels of iron in the diet may enhance tumor growth, and this should be considered when treating patients with cancer.

We reviewed conventional and chemical shift MR images and histologic findings of seven proven primary hepatic masses that had higher signal than liver on T1-weighted images to determine if this necessarily indicates fat and if the presence of fat indicates malignancy. These seven masses included five hepatocellular carcinomas (HCCs), one focal nodular hyperplasia (FNH), and one fatty dysplastic nodule. An eighth solitary high signal mass without histologic proof had evidence of abundant fat on each of two chemical shift MR images 25 months apart. Only one of the five HCCs had chemical shift or histologic evidence of fat, while the FNH and dysplastic nodule each had both chemical shift and histologic confirmation of fat. The dysplastic nodule became more dysplastic and grew significantly within 14 months, but remained benign. The unproven fatty lesion decreased in size over 25 months and is therefore presumably benign. Although no statistical inferences can be drawn from this small correlative study, we have shown that HCC may have higher signal intensity than liver on T1-weighted images, whether or not it contains fat. Chemical shift techniques can confirm the presence of intratumoral fat and thus indicate a mass of hepatocellular origin, but the mass may be benign or malignant.

The relationship of iron-binding proteins to prognosis was studied in 50 children at the Children's Hospital of Philadelphia, newly diagnosed with Hodgkin's disease (HD). There were five patients with Stage I, 18 with Stage II, 14 with Stage III, and 13 with Stage IV. Initial serum ferritin, transferrin, iron, hemoglobin (Hb), erythrocyte sedimentation rate (ESR), and A or B symptoms were analyzed for their association with progression-free survival (PFS). There was a linear increase of mean and median ferritin levels and a decrease of mean and median transferrin levels with advancing stages. Also, there was a significant inverse correlation between ferritin and transferrin (P less than 0.001). In univariate analyses, high ferritin (greater than 142 ng/ml) (P = 0.02) and low transferrin (less than or equal to 250 mg/dl) (P = 0.008) were significantly associated with poor PFS. Serum iron, Hb, ESR, and A or B symptoms were not associated with PFS. Stepwise proportional hazards regression analysis of all factors showed that transferrin was the only factor significantly associated with PFS. These preliminary results suggest that serum transferrin can also be used as a prognostic factor in addition to serum ferritin and that it may be helpful to assay both serum ferritin and transferrin as prognostic factors in childhood HD. Further testing of large groups of patients is needed to determine whether they are independent of tumor bulk and other established prognostic factors.

Changes in N-linked glycosylation are known to occur during the development of cancer. For example, increased branching of oligosaccharides has been associated with metastasis and has been correlated to tumor progression in human cancers of the breast, colon and melanomas. Increases in core fucosylation have also been associated with the development of hepatocellular carcinoma (HCC). Chronic infection with the hepatitis B virus is associated with more than 55% of all cases of hepatocellular carcinoma. We show here that increased levels of core fucosylation can be observed via glycan analysis of total serum and are associated with the development of HCC. In a blinded study, the serum glycoproteins derived from people diagnosed with HBV induced liver cancer were found to possess a dramatically higher level of fucosylation. This change occurs on both immunoglobulin molecules and on other serum glycoproteins. Targeted glycoproteomic analysis was used to identify those glycoproteins that are hyperfucosylated in cancer. In total, 19 proteins were found to be hyperfucosylated in cancer. The potential of these proteins as biomarkers of cancer is discussed.

Hepatitis B virus (HBV) carriers are at high risk for the development of hepatocellular carcinoma (HCC), but there are no reliable markers that will identify such high-risk carriers. The objective of this work is to identify serologic markers that may indicate the early presence of HCC. Since HBV-encoded X antigen (HBxAg) likely contributes to HCC by up- or down-regulation of host gene expression, X positive and negative HepG2 cells were made and subjected to cDNA subtraction. When specific ELISAs were constructed measuring differentially expressed antigens and corresponding antibodies, antibodies to several differentially expressed genes were detected. In cross-sectional and longitudinal studies, antibodies were predominantly present in patients with HBV-associated cirrhosis and HCC, but not in most carriers with hepatic inflammation alone or without active liver disease. Antibodies were also present in patients with hepatitis C virus (HCV)-related HCC, but rarely detected in sera from uninfected individuals, those with tumors other than HCC, or those with drug-induced hepatitis. Statistical analysis showed that HCC patients with four or more antibodies detectable before the appearance of HCC had decreased survival, suggesting that these markers may reflect stepwise hepatocarcinogenesis. Hence, these antibodies may serve as preneoplastic markers for HCC in HBV carriers with chronic liver disease, and may be identified by a simple blood test.

Hepatocellular carcinoma (HCC) is a major cause of morbidity and mortality, accounting for approximately 600,000 deaths annually world-wide and 10,000 deaths annually in the United States. Chronic infection with the hepatitis C virus (HCV) is the leading risk factor for HCC in the United States, and the second leading risk factor for HCC world-wide. The addition to the anti-HCV treatment armamentarium of boceprevir and telaprevir in 2011, and of simeprevir and sofosbuvir in 2013, mark a transition to a new era. In this new era, treatment for HCV is becoming so highly effective and tolerable that for the first time eradication of chronic HCV on a population level is realistically foreseeable. A major reduction in the burden of chronic HCV is likely to translate into a substantial reduction in the incidence of HCC. Here, we review the epidemiology, pathogenesis, and treatment of HCC with a special focus on hepatocarcinogenesis associated with chronic HCV infection.

As a preliminary study, we measured the necrosis of advanced hepatocellular carcinoma (HCC) by volume histogram after systemic chemotherapy and correlated it with clinical data. Five patients with advanced HCC secondary to chronic hepatitis and cirrhosis underwent pretreatment and posttreatment MR examination on a 1.5 T MR scanner following systemic chemotherapy. MR sequences included dynamic enhanced fast spoiled gradient echo 3D images. Clinical response to chemotherapy, as determined by MR images, was measured as changes of both the total tumor volume and the percent of tumor necrosis by volume histogram algorithm. Four of five patients had clinical improvement. Three of these patients had no or minimal change of tumor volume; however, there was an increase in tumor necrosis in follow-up MR image. One patient of five with no change in tumor necrosis had no response and died at 3 months. Serial MR images showed increased irregular necrosis of advanced HCC after systemic chemotherapy, but stable volume, in patients who responded clinically to systemic chemotherapy.

Hepatitis B virus (HBV) infection after liver transplantation (LT) may lead to severe and rapidly progressive graft failure. Antiviral treatment may be of benefit in selected patients with recurrent hepatitis B post-LT. The aim of this prospective open-label study is to determine the safety and efficacy of lamivudine in 33 liver transplant recipients with active HBV infection. The median time from LT to study enrollment was 51 months, all patients were hepatitis B surface antigen positive, and 75% and 94% of subjects had detectable hepatitis B e antigen (HBeAg) and HBV DNA at entry, respectively. The median duration of lamivudine treatment on study was 85 weeks, during which time median HBV DNA levels became undetectable by 16 weeks and 9% of patients lost previously detectable HBeAg. Serum alanine aminotransferase (ALT) levels improved in most patients and normalized in 27% of patients with elevated values pretreatment. Serum bilirubin and albumin levels significantly improved in patients with abnormal values at entry (P < .05). Virological breakthrough was detected in 13 subjects after a median of 61 weeks of lamivudine treatment and was confirmed to be caused by YMDD mutants in all patients tested. None of the patients with virological breakthrough showed a complete loss of clinical response to lamivudine. Serum ALT and bilirubin levels in patients with and without virological breakthrough were not significantly different at last study follow-up. Study results show that lamivudine is safe and effective in liver transplant recipients with recurrent hepatitis B. However, the high rate of virological breakthrough with prolonged therapy indicates the need for further studies of combination antiviral therapy in this patient population. Our results and others further establish the improving long-term outcomes with LT for patients with hepatitis B through advances in prevention of reinfection, as well as the availability of safe and effective antiviral therapies to treat patients with HBV recurrence. (Liver Transpl 2001;7:504-510.)

To investigate the magnetic resonance (MR) imaging findings of hepatic cirrhosis and the potential of MR imaging in differentiating cirrhosis from other hepatic abnormalities, three observers with different levels of expertise in MR imaging (specialist, experienced radiologist, and novice with special training) reviewed hepatic MR imaging examinations of 52 patients with biopsy-proved presence (n = 29) or absence (n = 23) of cirrhosis. All examinations included motion-compensated T1-weighted, T2-weighted, and flow-sensitive gradient-echo images. For all three observers, linear signal irregularity was more accurate than other findings. For the final diagnosis of cirrhosis, the specialist was most sensitive (76% at high threshold, 97% at low threshold), followed by the novice with special training (31% and 79%, respectively). Specificity was 100% for all observers at high threshold and 78%, 96%, and 87% for expert, experienced, and trained novice observers, respectively, at low threshold. Sensitivity did not vary regardless of severity of fibrosis, as determined independently by a hepatopathologist. MR imaging has the potential of offering a comprehensive noninvasive evaluation of patients with suspected cirrhosis, but considerable expertise is required at present.

To investigate the effects of iron supplementation on hepatoma cell growth, cells from a human hepatoma cell line, PLC/PRF/5, were grown in RPMI 1640 supplemented with 0, 10 and 20 micrograms/ml of FeSO4 and harvested weekly. At the end of 6 wk culture, cell mass measured 9.6, 14.7 and 13.2 gm, respectively. Amounts of ferritin from these cell masses were 0 (undetectable), 0.89 and 2.27 micrograms/gm of cells. To study the effects of iron deprivation of hepatoma cells, three human hepatoma cell lines (PLC/PRF/5, Hep G2 and Hep 3B) were incubated in tissue culture medium mixed with graded amounts of an iron-chelating agent, desferoxamine, for 48 to 96 hr at 37 degrees C with 5% CO2. Over 50% cell death in PLC/PRF/5 cells and 30% to 50% cell death in Hep G2 and Hep 3B cells were observed 48 to 72 hr after exposure to desferoxamine. Addition of ferric citrate partially reversed the cytotoxic effect of desferoxamine. On the other hand, viability of control cells, human diploid cell line (WI 38), was not affected by desferoxamine. Even after 96 hr exposure to desferoxamine, cell death was only 2% to 4%. These results suggest that (a) iron enhances tumor cell growth, (b) iron induces increased ferritin synthesis by tumor cells in vitro and (c) iron depletion causes tumor cell death but has little effect on normal human diploid cells. These findings should be considered when designing treatment of patients with hepatoma. Iron oversupply in patients with cancer might enhance tumor growth and adversely affect cancer therapy. Iron chelation with desferoxamine might have a place in the treatment of patients with hepatoma in conjunction with other anticancer agents.

A 10-year-old boy, who had been in an uninterrupted remission of acute lymphocytic leukemia (ALL) for six years, developed polycythemia vera (PV). One and a half months after detection of PV, he was found to have active leukemia. Both the polycythemia and leukemia receded with anti-leukemia therapy. Three possible explanations for the development of PV in a child with ALL are discussed: 1) PV was a part of his original ALL and recurred when patient relapsed. The PV phase was detected only during relapse because the patient was under close observation. 2) PV was a second neoplasm independent of ALL. 3) PV was part of a second leukemia which was different from the original leukemia; this new ALL was derived from a pluripotential cell line involving both erythroid and lymphoid elements. A precedent for this explanation has been observed in chronic myelogenous leukemia.

Serum ferritin levels are often elevated in patients with certain cancers and these elevations are, in part, derived from the tumors. In such patients, the increased levels of serum ferritin are associated with a poor prognosis. This association may be explained in part by biological effects of ferritin on lymphocytes: inhibition of E-rosette formation, masking of cell surfaces and suppression of lymphocytes' response to mitogens in vitro. The authors hypothesized that ferritins from tumor tissues also exert adverse effects on human granulocytes that are involved in tumoricidal activity. Three granulocyte functions were tested: nitroblue tetrazolium test, phagocytosis, and production of hydrogen peroxide. The results supported the authors' hypothesis: NBT reduction and phagocytosis are decreased in granulocytes exposed to ferritins, more so with tumor ferritins, than normal ferritin, and H2O2 production is less in granulocytes previously exposed to ferritins from tumor and nontumor tissues than cells not exposed to ferritins. However, the inhibitory effects of ferritins on H2O2 production can be reversed if granulocytes are further stimulated by phorbol myristate acetate (a membrane stimulant). If the elevated serum ferritin in cancer patients impairs granulocyte functions, in vivo, then it may increase the risk of infection, decrease tumoricidal host responses, and, thereby, contribute to the poor prognosis of these individuals.

Previous studies from this laboratory support the view that increased serum ferritin levels are associated with an increased risk of primary hepatocellular carcinoma (PHC). We have tested this hypothesis in a population of Korean patients with chronic liver disease followed for development of PHC. Serum ferritin levels were measured over time in 249 patients with liver diseases (mostly chronic) followed for 2 to 17 years in Seoul, Korea. Most of the patients were chronically infected with hepatitis B virus. During the first 8 months of follow-up, there were no cases of PHC and no deaths. During this same period, no patient had a serum ferritin level initially below 300 ng/ml and rising above 300 ng/ml, but some patients with ferritin levels above 300 ng/ml experienced decreases to below 300 ng/ml. Therefore, patients were grouped by ferritin level during the first 8 months of follow-up into 3 categories according to the above criteria. Multivariate analysis showed that consistently elevated ferritin levels (category 3) were significantly associated with the development of PHC. Men were more likely to have elevated ferritin levels than women and were at higher risk of developing PHC. Men who were chronically infected with HBV and had ferritin levels above 300 ng/ml had a 50% chance of developing PHC during the follow-up period, compared with a 20% risk of PHC for men with lower ferritin levels (categories 1 and 2). This elevated risk of PHC in men with elevated ferritin levels was confined to the first 3 years of follow-up.

Serum ferritin frequently is elevated in patients with neuroblastoma. Isoferritins extracted from neuroblastoma tumors and cells in culture show a wide range from basic (rich in L subunit) to acidic (rich in H subunit) isoferritins. Total ferritin is a combination of basic and acidic isoferritins. Forty-four serum samples from 25 patients with neuroblastoma were measured for basic and acidic isoferritin levels by radioimmunoassay using antibodies to liver (basic) ferritin and HeLa (acidic) ferritin. Normal ranges for basic and acidic serum ferritins were 7 to 142 ng/ml (median, 30 ng/ml) and 0 to 12 ng/ml (median, 3.4), respectively. Basic ferritins in the 44 neuroblastoma sera ranged from 0 to 1460 ng/ml, and acidic ferritins, 0 to 40 ng/ml. Sera with high levels of acidic ferritins always had increased basic ferritins. Thus, acidic/basic ferritin ratios were nearly constant, less than 0.3 in all sera. There was a significant linear correlation between basic and acidic isoferritins (r = 0.833). These results suggest that neuroblastoma tumors produce both basic and acidic isoferritins and release them into circulation. However, there is no acidic ferritinemia not accompanied by basic ferritinemia. Therefore, the commercial assay for basic isoferritin currently seems sufficient for clinical prognostic purposes.

Known prognostic factors in neuroblastoma were analyzed in 124 children to determine which were independent and which were most useful in predicting outcome. The following factors were analyzed: age, sex, stage of disease, serum neuron-specific enolase (NSE), serum ferritin, E-rosette inhibition, urinary catecholamines, and histologic type according to the criteria of Shimada. Estimates of survival were calculated using the method of Kaplan and Meier. The overall survival for 124 patients was 60% at 2 years. There were significant differences in survival by pathology, age, NSE, ferritin, vanilmandelic acid (VMA): homovanillic acid (HVA) ratio, and stage. There was a strong association among NSE, age, stage, and ferritin. Using the recursive partitioning approach, it was possible to subdivide patients into three groups (based on diagnostic values of ferritin, age, and stage) with a good, intermediate, and poor prognosis and estimated 2-year survival of 100%, 62%, and 19%, respectively. Further analysis could not be done because of small numbers in the subgroups, but the results suggest that combinations of age, stage, serum ferritin, and histologic type may be able to define two populations: favorable with 80% + 2-year survival and unfavorable with less than 20%.

Lymphoblasts from 23 children with acute lymphocytic leukemia (ALL) and 10 with lymphoblastic lymphoma (LBL) were studied by complement-dependent microcytoxicity tests with two nonhuman primate antisera defining leukemia-associated and lymphoma-associated antigens. Cells form 15 patients with ALL and 1 with LBL reacted only with antiserum to chronic lymphatic leukemia (CLL). These group-I patients were predominantly female. Most were pancytopenic and lacked mediastinal widening and T-cell markers; lymphoblasts from 15 were periodic acid-Schiff-positive. Cells from 8 male patients reacted only with antiserum to converted lymphosarcoma (LS). All these group-II patients expressed T-cell markers; 5 had mediastinal enlargement and 2, an abdominal mass. Six of the 8 were PAS-negative. Cells from 9 patients reacted with both antisera. The group-III patients demonstrated some characteristics of each of the above groups. Patients whose lymphoblasts reacted with CLL antiserum presented with clinical and laboratory features indicative of a good prognosis, i.e., ALL with PAS positivity and no T-cell markers or localized mass. Patients whose cells reacted with LS antiserum often had bad prognostic features: mediastinal or abdominal mass, expression of T-cell markers, and PAS negativity. These antisera appear able to differentiate childhood ALL from LBL. The distinction is important prognostically and perhaps therapeutically.

The distribution of T, B and null lymphocytes was studied in the peripheral blood (PB) of 34 children with acute lymphocytic leukemia (ALL), at diagnosis and during the course of their disease. All patients received the same chemotherapy. At diagnosis, the proportion and absolute numbers of T lymphocytes fell into 2 groups, a "low" group (medians 8%, 318/mm3) and a "normal" group (medians 60%, 1405/mm3). Patients with a low proportion of T cells tended to have a high proportion and absolute number of null cells (correl. coeff. r = 0.94). This group has had high white blood cell counts (median = 29,600/mm3) and a high proportion of blasts (84%). During remission, the proportion of T cells returned toward normal (40-75%) in both groups. Four patients in the low T group relapsed; T cells again fell to low levels and returned to normal in response to new therapy. Three interpretations of these observations are suggested: 1) Active leukemia may cause normal lymphocytes (T cells and to a lesser degree B cells) to lose their surface markers; 2) Acute leukemia may inhibit maturation of normal lymphoid cells, and thus T or B markers are not sufficiently developed to be detectable; and 3) Cells without surface markers (null cells) may be a part of the malignant population, even though they are not morphologic blasts. Further studies are necessary to test these hypotheses.