Unique intrahepatic transcriptomics profiles discriminate the clinical phases of a chronic HBV infection

Abstract

Chronic hepatitis B is a highly heterogeneous liver disease characterized by phases with fluctuations in viral replication and progressive liver damage in some, but not all infected individuals. Despite four decades of research, insight into host determinants underlying these distinct clinical phases—immunotolerant, immune active, inactive carrier, and HBeAg-negative hepatitis–remains elusive. We performed an in-depth transcriptome analysis of archived FFPE liver biopsies of each clinical phase to address host determinants associated with the natural history. Therefore, we determined, for the first time, intrahepatic global expression profiles of well-characterized chronic HBV patients at different clinical phases. Our data, obtained by microarray, demonstrate that B cells and NK/cytotoxic-related genes in the liver, including CD19, TNFRSF13C, GZMH, and KIR2DS3, were differentially expressed across the clinical HBV phases, which was confirmed by modular analysis and also Nanostring arrays in an independent cohort. Compared to the immunotolerant phase, 92 genes were differentially expressed in the liver during the immune active phase, 46 in the inactive carrier phase, and 71 in the HBeAg-negative phase. Furthermore, our study also revealed distinctive transcription of genes associated with cell cycle activity, NF-κB signaling, cytotoxic function and mitochondrial respiration between clinical phases. Our data define for the first time using microarray unique transcriptomes in the HBV-infected liver during consecutive clinical phases. We demonstrate that fluctuations of viral loads and liver damage coincide with fluctuations in the liver transcriptome and point to functional- immune and non-immune- components contributing to the clinical phenotype in patients.

Hepatitis B core-specific memory B cell responses associate with clinical parameters in patients with chronic HBV

“…[15][16][17] We have recently shown that at the transcriptome level, B cell-related genes are expressed more in disease phases with elevated ALT, both in blood and livers of untreated patients with CHB, suggesting a direct or indirect involvement in the immunopathogenesis. 1,9 Here we applied fluorescently labeled HBcAg as baits to ex vivo track HBcAg-specific B cells in blood samples of a wellcharacterized cohort of patients with CHB. We also examined functional differences in B cell ELISPOT assays.…”

“…Serum and/or peripheral blood mononuclear cells (PBMCs) were obtained from patients with CHB (n = 152, Table 1) attending the outpatient clinic of the Erasmus Medical Center, Rotterdam, the Netherlands or the Antwerp University Hospital, Antwerp, Belgium and categorized into 4 clinical HBV phases: immune tolerant (IT) or "HBeAg-positive chronic infection" -HBeAg+, HBeAb-, repetitive normal ALT (<upper limit of normal [ULN]); immune active (IA) or "HBeAg-positive chronic hepatitis" -HBeAg+, repetitive or intermittent high ALT (>ULN); inactive carrier (IC) or "HBeAg-negative chronic infection" -HBeAg-, HBeAb+, low ALT, HBV DNA <20,000 IU/ml; HBeAg-hepatitis (ENEG) or "HBeAg-negative chronic hepatitis" -HBeAg-, HBeAb+, repetitive or intermittent high ALT (>ULN). Categorization was performed as previously described 1,9,11 with minor modifications: i) no restrictions regarding the level of liver fibrosis, although most patients had < − F2 fibrosis; ii) cohorts of HBsAg-vaccinated healthy controls (HCs, n = 23) and virally suppressed nucleos(t) ide analogue (NUC)-treated patients with CHB (n = 34) were included; iii) For some IC patients intermittent ALT < − 2 ULN was allowed if there was documented liver steatosis without histological steatohepatitis and no other signs of liver disease. 12 ALT levels of IC patients remained significantly lower compared to ENEG and IA patients (Table 1; p <0.001).…”

“…Second, we have previously identified B cell-related transcriptomic changes in blood and liver of patients with CHB paralleling clinical parameters in subsequent HBV disease phases. 1,9 HBsAg-specific B cell studies do not reflect this dichotomized disease presentation of CHB, as the numbers or function of HBsAg-specific B cells remain stable irrespective of HBV DNA or alanine aminotransferase (ALT) levels.…”

Stopping nucleos(t)ide analogue treatment in Caucasian hepatitis B patients after HBeAg seroconversion is associated with high relapse rates and fatal outcomes

“…So far published studies assessing the clinical consequences of NA treatment stop after HBeAg seroconversion mainly focus on Asian patients . However, it is well appreciated that the host's ethnicity influences HBV's natural history and immune responses . Asian patients show, for example, lower spontaneous or treatment‐induced HBsAg seroclearance rates, and have higher HBV relapse rates in HBeAg‐negative patients after treatment cessation .…”

“…12 However, it is well appreciated that the host's ethnicity influences HBV's natural history and immune responses. 4,15 Asian patients show, for example, lower spontaneous or treatmentinduced HBsAg seroclearance rates, and have higher HBV relapse rates in HBeAg-negative patients after treatment cessation. 16 Clinical hepatocellular carcinoma risk scores developed in Asian cohorts seem to be unreliable in Caucasian and especially African patients, the latter developing hepatocellular carcinoma at much younger ages.…”