Hiroiku Kawakami

Circulating microRNA‐22 correlates with microRNA‐122 and represents viral replication and liver injury in patients with chronic hepatitis B

Hepatitis B virus (HBV) infection is associated with increased expression of microRNA‐122. Serum microRNA‐122 and microRNA‐22 levels were analyzed in 198 patients with chronic HBV who underwent liver biopsy and were compared with quantitative measurements of HBsAg, HBeAg, HBV DNA, and other clinical and histological findings. Levels of serum microRNA‐122 and microRNA‐22 were determined by reverse transcription‐TaqMan PCR. Serum levels of microRNA‐122 and microRNA‐22 were correlated (R2 = 0.576; P < 0.001), and both were elevated in chronic HBV patients. Significant linear correlations were found between microRNA‐122 or microRNA‐22 and HBsAg levels (R2 = 0.824, P < 0.001 and R2 = 0.394, P < 0.001, respectively) and ALT levels (R2 = 0.498, P < 0.001 and R2 = 0.528, P < 0.001, respectively). MicroRNA‐122 levels were also correlated with HBV DNA titers (R2 = 0.694, P < 0.001 and R2 = 0.421, P < 0.001). Levels of these microRNAs were significantly higher in HBeAg‐positive patients compared to HBeAg‐negative patients (P < 0.001 and P < 0.001). MicroRNA‐122 levels were also lower in patients with advanced liver fibrosis (P < 0.001) and lower inflammatory activity (P < 0.025). These results suggest that serum micro‐RNA levels are significantly associated with multiple aspects of HBV infection. The biological meaning of the correlation between microRNA‐122 and HBsAg and should be investigated further. J. Med. Virol. 85:789–798, 2013.

Serum HBV RNA is a predictor of early emergence of the YMDD mutant in patients treated with lamivudine

Lamivudine (LAM) is a nucleoside analogue widely used for the treatment of chronic hepatitis B virus (HBV) infection. Emergence of resistant strains with amino acid substitutions in the tyrosine‐methionine‐aspartate‐aspartate (YMDD) motif of reverse transcriptase is a serious problem in patients on LAM therapy. The amount of covalently closed circular DNA in the serum is reported to be higher in patients who develop YMDD mutants than in those without mutants. However, there is no useful serum marker that can predict early emergence of mutants during LAM therapy. Analysis of patients who were treated with entecavir (n = 7) and LAM (n = 36) showed some patients had high serum levels of HBV RNA. Median serum levels of HBV RNA were significantly higher in patients in whom the YMDD mutant had emerged within 1 year (n = 6, 1.688 log copies/ml) than in those in whom the YMDD mutant emerged more than 1 year after treatment (n = 12, 0.456 log copies/ml, P = 0.0125) or in whom the YMDD mutant never emerged (n = 18, 0.688 log copies/ml, P = 0.039). Our results suggest that HBV RNA is a valuable predictor of early occurrence of viral mutation during LAM therapy. (HEPATOLOGY 2007;45:1179–1186.)

Effect of vitamin D supplementation on pegylated interferon/ribavirin therapy for chronic hepatitis C genotype 1b : A randomized controlled trial

Chronic HCV–infected patients tend to have vitamin D deficiency, suggesting that vitamin D supplementation may enhance the efficacy of treatment with pegylated interferon (PEG‐IFN) and ribavirin (RBV). We therefore assessed the effects of vitamin D supplementation on viral response to PEG‐IFN/RBV. Eighty‐four patients with HCV genotype 1b were randomized, 42 to oral vitamin D supplementation (1000 IU/day) and 42 to nonsupplementation (control), from week 8 to the end of PEG‐IFN/RBV therapy. The primary end point was undetectable HCV RNA at week 24 (viral response [VR]). VR rate at week 24 was significantly higher in the vitamin D than in the control group (78.6% vs 54.8% P = 0.037). Adverse events were similar in both groups. When patients were subdivided by IL28B SNP rs8099917 genotype, those with the TT genotype group showed a significantly higher VR rate at week 24 with than without vitamin D supplementation (86.2% vs 63.3% vs P = 0.044). Although patients with the TG/GG genotype, who were relatively resistant to PEG‐IFN treatment, had similar VR rates at week 24 with and without vitamin D supplementation, the decline in viral load from week 8 to week 24 was significantly greater with than without vitamin D supplementation. Multivariate analysis showed that rs8099917 genotype and vitamin D supplementation contributed significantly to VR at week 24. SVR rates were similar in the vitamin D and control groups [64.3% (27/42) vs 50% (21/42), P = 0.19]. Vitamin D supplementation may enhance the effects of PEG‐IFN/RBV in HCV genotype 1b–infected patients.

Eicosapentaenoic Acid Could Permit Maintenance of the Original Ribavirin Dose in Chronic Hepatitis C Virus Patients during the First 12 Weeks of Combination Therapy with Pegylated Interferon-α and Ribavirin

Objective: To evaluate the efficacy of eicosapentaenoic acid (EPA) against ribavirin (RBV)-associated hemolytic anemia during the first 12 weeks in chronic hepatitis C virus (HCV) combination therapy. Methods: This study was a prospective open-label, randomized controlled trial. 100 HCV patients were randomized to either the EPA group (n = 49) or non-EPA group (n = 51) who received combination therapy with or without EPA. We compared the changes in hemoglobin level and RBV plasma concentrations at week 12 in each group with RBV dose reduction rate and performed multivariate analysis to identify independent variables associated with RBV dose reduction. Results: 8 patients (17%) in the EPA group and 20 patients (29%) in the non-EPA group required RBV dose reduction, respectively. The cumulative RBV reduction rate was significantly lower in the EPA group than in the non-EPA group (p = 0.017), while the decrease of hemoglobin and RBV plasma concentrations from baseline was not significantly different. However, in the multivariate analysis, treatment with EPA showed significant variables for the reduction of RBV dose (odds ratio 3.235, p = 0.023). Conclusion: EPA could prevent the RBV dose reduction during the first 12 weeks in combination therapy, although further large-scale double-blind randomized controlled trials are required.

Therapeutic Effect of a Low Dosage of Human Leukocyte Interferon on Chronic Hepatitis B Virus Infection

A low dosage of human leukocyte interferon was intramuscularly given to 47 patients with hepatitis B surface antigen (HBsAg) positive chronic active hepatitis once a week for 4 consecutive weeks (10 X 10(5), 5 X 10(5), 2 X 10(5) and 1 X 10(5) U). After treatment, a reduction of serum HBsAg was observed in 26 patients; 3 of them showed no serum HBsAg and 1 of the 3 appeared to produce antibody to HBsAg. 31 of 34 patients investigated showed a significant decrease in Dane particle-associated DNA-polymerase activity (p less than 0.001). Among 17 patients positive for hepatitis B e antigen (HBeAg), 10 of them seroconverted to anti-HBe. Serum transaminase levels also significantly improved in 34 of 38 patients (p less than 0.001). Our findings indicate that a low dosage of human leukocyte interferon such as 18 X 10(5) U, administered in progressively decreasing doses, may be effective in the treatment of chronic hepatitis B virus infection.

Dose comparison study of pegylated interferon-α-2b plus ribavirin in naïve Japanese patients with hepatitis C virus genotype 2: A randomized clinical trial

Background and Aim:  To compare the efficacy and safety of pegylated interferon (PEG‐I) at 1 and 1.5 µg/kg, and in combination with ribavirin (RBV) for 24 weeks in naïve Japanese patients infected with hepatitis C virus genotype 2.

Prospective study of short‐term peginterferon‐α‐2a monotherapy in patients who had a virological response at 2 weeks after initiation of interferon therapy

Background and Aims:  Long‐term interferon (IFN) therapy is effective in eliminating hepatitis C virus (HCV). However, it carries the risk of adverse effects and reduced quality of life. To assess whether short‐term IFN therapy effectively eliminates HCV, we performed a prospective pilot study of pegylated (peg)IFN‐α‐2a therapy for 8 or 24 weeks.

Impact of ribavirin dose reduction on the efficacy of pegylated interferon plus ribavirin combination therapy for elderly patients infected with genotype 1b and high viral loads

Aim:  To examine the impact of ribavirin dose reduction on the efficacy of pegylated interferon (PEG IFN) plus ribavirin combination therapy for elderly patients infected with genotype 1b and high viral loads.

Defective suppressor function of spleen cells by neonatal thymectomy and persistence of experimental auto‐immune hepatitis in mice

The effects of neonatal thymectomy on a model of auto‐immune hepatitis prepared by immunizing A/J mice with syngeneic crude liver proteins were studied. Hepatitis was more severe in neonatally thymectomized mice (Group B) than in non‐thymectomized controls (Group A). Changes in the hepatic lobules were reduced in both groups 3 months after the final immunization, but inflammation around the portal areas tended to persist in Group B; only this group showed frequent infiltration of mononuclear cells accompanied by destruction of the limiting plate in the portal area. The serum level of auto‐antibody to liver‐specific membrane lipoprotein complex (LSP) and delayed‐type hypersensitivity (DTH) to LSP were higher in Group B than in Group A. The auto‐antibody to LSP was positive in both groups 3 months after the final immunization. Although it was significantly reduced in Group A compared with the level 3 days after the final immunization, it remained high in Group B. Liver damage, production of the auto‐antibody to LSP and DTH to LSP were reduced by adoptive transfer of normal murine spleen cells, but the spleen cells of neonatally thymectomized mice showed no such effects. These results suggest that normal spleen cells possess the capability of suppressing auto‐immune hepatitis, and the defect of this capability due to T‐cell dysfunction by neonatal thymectomy contributes to the promotion and persistence of auto‐immune hepatitis.

Prolonged Negative HCV-RNA Status Led to a Good Outcome in Chronic Hepatitis C Patients with Genotype 1b and Super-High Viral Load

Objective: We examined whether a sustained negative HCV-RNA status for 48 weeks affects the outcome in patients with genotype 1b and super-high viral load, and also investigated whether the outcome is affected by the induction therapy of twice-daily pre-administrated interferon (IFN)-β. Methods: 78 eligible patients were divided into four groups. 40 were patients assigned to the short treatment protocol. 13 patients received 3 MU IFN-β twice daily for 2 weeks followed by IFN-α2b+ribavirin for 22 weeks (β-induction group: group 1). 27 patients received IFN-α2b+ribavirin for 24 weeks (standard combination group: group 2). 38 patients were assigned to the maintenance treatment protocol. All of the 13 in the β-induction group (group 3) and 21 of 25 patients in the standard combination group (group 4) who were negative HCV-RNA PCR at week 24 had IFN monotherapy to maintain a negative HCV-RNA result for 48 weeks. Results: An HCV-RNA-negative status at week 24 was observed in 96% (25/26) of groups 1 and 3 versus in 79% (41/52) of groups 2 and 4 (p < 0.01). The sustained virological response (SVR) was 38% (5/13) in group 1 and 11% (3/27) in group 2 (p < 0.05). In the maintenance treatment, SVR was observed in 46% (6/13) of group 3 and 32% (8/25) of group 4 (NS). Conclusions: A sustained negative HCV-RNA status for 48 weeks might be associated with viral elimination in patients with genotype 1 and super-high viral load.