A serum “sweet-doughnut” protein facilitates fibrosis evaluation and therapy assessment in patients with viral hepatitis

Abstract

Although liver fibrosis reflects disease severity in chronic hepatitis patients, there has been no simple and accurate system to evaluate the therapeutic effect based on fibrosis. We developed a glycan-based immunoassay, FastLec-Hepa, to fill this unmet need. FastLec-Hepa automatically detects unique fibrosis-related glyco-alteration in serum hyperglycosylated Mac-2 binding protein within 20 min. The serum FastLec-Hepa counts increased with advancing fibrosis and illustrated significant differences in medians between all fibrosis stages. FastLec-Hepa is sufficiently sensitive and quantitative to evaluate the effects of PEG-interferon-α/ribavirin therapy in a short post-therapeutic interval. The obtained fibrosis progression is equivalent to -0.30 stages/year in patients with sustained virological response, and 0.01 stages/year in relapse/nonresponders. Furthermore, long-term follow-up of the severely affected patients found hepatocellular carcinoma developed in patients after therapy whose FastLec-Hepa counts remained above a designated cutoff value. FastLec-Hepa is the only assay currently available for clinically beneficial therapy evaluation through quantitation of disease severity.

Molecular Basis for Recognition of the Cancer Glycobiomarker, LacdiNAc (GalNAc[β1→4]GlcNAc), by Wisteria floribunda Agglutinin

“…Thus, its cross-reactive potential severely limits its use for cancer biomarker detection. WFA is currently considered the most prominent diagnostic lectin against cholangiocarcinoma when compared with other homologous lectins (26,35) and the most discriminatory against altered N-glycans on Mac2-binding protein, a secretory N-glycoprotein with elevated expression levels during viral hepatitis induced liver cirrhosis (36,37).…”

Clinical Evaluation of Hepatocarcinogenesis and Outcome Using a Novel Glycobiomarker <i>Wisteria floribunda</i> Agglutinin-Positive Mac-2 Binding Protein (WFA⁺-M2BP) in Chronic Hepatitis C with Advanced Fibrosis

“…The glycan-based immunoassay LecT-Hepa is a previously developed simple assay system that automatically detects unique fibrosis-related glyco-alterations (24). WFA + -M2BP was recently confirmed as a liver fibrosis glycobiomarker that can be assessed using a fully automated immunoassay (8). The different fibrosis stages can be readily verified by checking serum WFA + -M2BP levels (25).…”

“…Routine measurements of serum HCV RNA were performed using the Cobas AmpliPrep/Cobas TaqMan HCV test version 2.0 (Roche Diagnostics K.K., Tokyo, Japan). Serum WFA + -M2BP was quantified using a lectin-antibody sandwich immunoassay on a fully automatic HISCL2000i immunoanalyzer (Sysmex Co., Kobe, Japan) (8). In this study, according to a previous report (13), serum WFA + -M2BP levels were classified into 3 groups: high (≥ 4 cutoff index [COI]), intermediate (1-4 COI), or low (< 1 COI) levels.…”

“…WFA is thought to recognize the GalNAc residues of N-glycans and O-glycans and the clustered LacNAc structure (9). An automatic and high-throughput assay can measure WFA + -M2BP levels in 10-μL serum samples (8). The WFA + -M2BP level in CHC patients was found to increase with the progression of liver fibrosis (10) and was found to be a predictor of HCC (11).…”

“…Glycoprotein-based biomarkers (glycobiomarkers) are novel recently developed biomarkers of disease. In 2013, the Mac-2 binding protein glycosylation isomer (also called Wisteria floribunda agglutinin-positive Mac-2 binding protein [WFA + -M2BP]) was introduced as a novel, noninvasive, rapidly assayed serological glycobiomarker for evaluating liver fibrosis (8). M2BP has been found to have multibranching and sialylated N-glycans.…”