Discovery of a Potential Plasma Protein Biomarker Panel for Acute-on-Chronic Liver Failure Induced by Hepatitis B Virus.

Ni Zhou,Yongzhi Tang,Tingting Huang,Fei Yan,Huazhong Chen,Kuifeng Wang,Hu Zhou,Shanhua Fang,Xiaoyu Zhao,Jiansheng Zhu

doi:10.3389/fphys.2017.01009

Abstract

Hepatitis B virus (HBV)-associated acute-on-chronic liver failure (HBV-ACLF), characterized by an acute deterioration of liver function in the patients with chronic hepatitis B (CHB), is lack of predicting biomarkers for prognosis. Plasma is an ideal sample for biomarker discovery due to inexpensive and minimally invasive sampling and good reproducibility. In this study, immuno-depletion of high-abundance plasma proteins followed by iTRAQ-based quantitative proteomic approach was employed to analyze plasma samples from 20 healthy control people, 20 CHB patients and 20 HBV-ACLF patients, respectively. As a result, a total of 427 proteins were identified from these samples, and 42 proteins were differentially expressed in HBV-ACLF patients as compared to both CHB patients and healthy controls. According to bioinformatics analysis results, 6 proteins related to immune response (MMR), inflammatory response (OPN, HPX), blood coagulation (ATIII) and lipid metabolism (APO-CII, GP73) were selected as biomarker candidates. Further ELISA analysis confirmed the significant up-regulation of GP73, MMR, OPN and down-regulation of ATIII, HPX, APO-CII in HBV-ACLF plasma samples (p < 0.01). Moreover, receiver operating characteristic (ROC) curve analysis revealed high diagnostic value of these candidates in assessing HBV-ACLF. In conclusion, present quantitative proteomic study identified 6 novel HBV-ACLF biomarker candidates and might provide fundamental information for development of HBV-ACLF biomarker.

Highlights

Acute-on-chronic liver failure (ACLF) is increasingly recognized as an acute deterioration of liver function combining with liver and multi-organ failures in patients with pre-existing chronic liver disease, there is no consensus about its definition (Bernal et al, 2015; Anand and Dhiman, 2016; Arroyo and Jalan, 2016)
Plasma samples from healthy control, chronic hepatitis B (CHB) and Hepatitis B virus (HBV)-ACLF patients were subjected to LC-mass spectrometry (MS)/MS analysis following removal of high abundance protein, Filter Assisted Sample Preparation (FASP) preparation, tryptic digestion and isobaric tagging for relative and absolute quantitation (iTRAQ) labeling
Most researchers agree that the concept of ACLF should include: acute deterioration of preexisting chronic liver disease, multi-system organ failure and with a mortality ≥15% at day 28 (Kim and Kim, 2013; BlascoAlgora et al, 2015)

Summary

Introduction

Acute-on-chronic liver failure (ACLF) is increasingly recognized as an acute deterioration of liver function combining with liver and multi-organ failures in patients with pre-existing chronic liver disease, there is no consensus about its definition (Bernal et al, 2015; Anand and Dhiman, 2016; Arroyo and Jalan, 2016). It is believed that predicting and stopping the progression of CHB to ACLF at an early stage may be the most effective strategy of reducing the mortality of patients with HBV-ACLF. Wan et al (2015) performed a particleenhanced immunonephelometry assay on serum samples from ACLF patients, and they found that the level of cystain C (CysC) was significantly higher in the ACLF with kidney failure group than those in the healthy controls and CHB patients. Their results suggested that CysC could be considered as a biomarker for renal dysfunction in ACLF patients. Considering the complexity and heterogeneity of HBV-ACLF pathology, it has been suggested that integrated panel of biomarkers with specific and complementary functions rather than a single biomarker be useful in diagnosis of patients with HBV-ACLF

Methods

Results

Conclusion