Differential Plasma MicroRNA Profiles in HBeAg Positive and HBeAg Negative Children with Chronic Hepatitis B

Abstract

Background and AimChildren chronically infected with hepatitis B virus (HBV) are at high risk of progressive liver disease. However, no treatment is available that is consistently effective in curing chronic hepatitis B (CHB) in children. Improved understanding of the natural course of disease is warranted. Identification of specific microRNA (miRNA) profiles in children chronically infected with HBV may provide insight into the pathogenesis of CHB and lead to advances in the management of children with CHB.Patients and MethodsMiRNA PCR panels were employed to screen plasma levels of 739 miRNAs in pooled samples from HBeAg positive, HBeAg negative, and healthy children. The three groups’ plasma miRNA profiles were compared, and aberrantly expressed miRNAs were identified. The identified miRNAs were then validated. Individual RT-qPCRs were performed on plasma from 34 HBeAg positive, 26 HBeAg negative, and 60 healthy children.ResultsA panel of 16 plasma miRNAs were identified as aberrantly expressed in HBeAg positive and HBeAg negative children (p<0.001). Levels of all of the miRNAs were upregulated in HBeAg positive children compared with in HBeAg negative children. A positive correlation was furthermore found between plasma levels of the identified miRNAs and HBV DNA (p<0.001).ConclusionWe are the first to investigate the plasma miRNA profile of children chronically infected with HBV. Our data indicates the existence of a relationship between abundance of circulating miRNAs and immunological stages in the natural course of disease. Certain miRNAs may contribute to the establishment and maintenance of CHB in children. Further studies are warranted to advance understanding of miRNAs in the pathogenesis of CHB, hopefully leading to the identification of future therapeutic targets.

Differences in serum microRNA profiles in hepatitis B and C virus infection

“…However, Winther et al reported similar results in children with chronic hepatitis B and found that plasma levels of a subset of microRNAs decreased significantly in one child before and after HBe seroconversion. 50 We have previously shown that both HBc and HBs proteins colocalize and physically interact with AGO2 in hepatocytes and that siRNA ablation of AGO2 suppressed HBV DNA and HBsAg production, 10 suggesting that components of the RNA silencing machinery are recruited during HBV replication. HSP90 has been reported to act as a chaperone during RNA loading of Argonaute proteins 51 and is also essential in catalyzing HBV reverse transcription and capsid formation by interacting with the pregenomic RNA encapsidation signal, reverse transcriptase, and the core protein.…”

Survey of 800+ data sets from human tissue and body fluid reveals xenomiRs are likely artifacts

“…Over the past decade, RNAs extracted from cell-free body fluids have received increasing attention as novel diagnostic markers for disorders as diverse as narcolepsy, cancers, and viral infections (Mitchell et al 2008;Winther et al 2013;Holm et al 2014a). Many distinct body fluids have been profiled for biomarkers (Weber et al 2010;Holm et al 2014b).…”

Function and Regulation of MicroRNAs and Their Potential as Biomarkers in Paediatric Liver Disease

“…Thirty-two miRNAs were shown to be significantly differentially expressed between HBeAg+ and HBeAg− children. In addition to the 14 miRNAs described in the previous studies [147,148], nine upregulated and nine downregulated miRNAs were identified.…”

“…Children in the immune inactive stage have a lower risk of liver disease progression, however, hepatitis B virus reactivation may occur [173]. Winther et al compared plasma miRNA levels in children with HBeAg+ and HBeAg− versus healthy controls (Table 3) [147]. Differentially expressed miRNAs were assessed by PCR arrays, identifying the upregulation of 16 miRNAs (miRNA-99a, miRNA-100, miRNA-122-5p, miRNA-122-3p, miRNA-125b, miRNA-192-5p, miRNA-192-3p, miRNA-193b, miRNA-194, miRNA-215, miRNA-356, miRNA-455-5p, miRNA-455-3p, miRNA-483-3p, miRNA-885-5p and miRNA-1247) for which expression was highest in the HBeAg+ group, lower in HBeAg− group and lowest in the healthy control group.…”

“…Using the raw data from the original study [147], plasma miRNA expression was re-analyzed from HBeAg+, HBeAg− and the healthy control groups. Thirty-two miRNAs were shown to be significantly differentially expressed between HBeAg+ and HBeAg− children.…”