Elevated levels of circulating ITIH4 are associated with hepatocellular carcinoma with nonalcoholic fatty liver disease: from pig model to human study

Abstract

Background Noninvasive biomarkers are urgently needed for optimal management of nonalcoholic fatty liver disease (NAFLD) for the prevention of disease progression into nonalcoholic steatohepatitis (NASH) and hepatocellular carcinoma (HCC). In order to identify the biomarkers, we generated the swine hepatocellular carcinoma (HCC) model associated with NAFLD and performed serum proteomics on the model. Methods Microminipigs were fed a high-fat diet to induce NAFLD and a normal diet as the control. To induce HCC, diethylnitrosamine was intraperitoneally administered. Biopsied liver samples were histopathologically analyzed every 12 weeks. Serum proteins were separated by blue native two-dimensional gel electrophoresis and proteins of interest were subsequently identified by MALDI-TOF MS/MS. Human serum samples were analyzed to validate the candidate protein using antibody-mediated characterization. Results In the NAFLD pigs, hepatic histology of nonalcoholic steatohepatitis (NASH) was observed at 36 weeks, and HCC developed at 60 weeks. Among serum proteins identified with MALDI-TOF MS/MS, serum inter-alpha-trypsin inhibitor heavy chain 4 (ITIH4), an acute response protein which is secreted primarily by liver, was identified as the most characteristic protein corresponding with NAFLD progression and HCC development in the NAFLD pigs. With immunoassay, serum ITIH4 levels in the NAFLD pigs were chronologically increased in comparison with those in control animal. Furthermore, immunohistochemistry showed ITIH4 expression in hepatocytes also increased in both the cancer lesions and parenchyma as NAFLD progressed. Human study is also consistent with this observation because serum ITIH4 levels were significantly higher in HCC-NAFLD patients than in the simple steatosis, NASH, and virus-related HCC patients. Of note, HCC-NAFLD patients who had higher serum ITIH4 levels exhibited poorer prognosis after hepatectomy. Conclusions We established an HCC pig model associated with NAFLD. Serum proteomics on the swine HCC with NAFLD model implicated ITIH4 as a non-invasive biomarker reflecting NAFLD progression as well as subsequent HCC development. Most importantly, the results in the swine study have been validated in human cohort studies. Dissecting speciation of serum ITIH4 promises to have clinical utility in monitoring the disease. Electronic supplementary material The online version of this article (10.1186/s12885-019-5825-8) contains supplementary material, which is available to authorized users.

Alveolar Epithelial Inter-alpha-trypsin Inhibitor Heavy Chain 4 Deficiency Associated With Senescence-regulated Apoptosis by Particulate Air Pollution

“…Our previous report indicated that serum ITIH4 had a good correlation with serum CRP levels in COPD patients after PM 10 exposure. Circulating ITIH4 may be associated with liver regulation into the systemic circulation as reported previously [14,15], and it was also correlated with circulating CRP levels. If lung-speci c ITIH4 has the same function as the acute-phase response, it may be associated with the in ammatory response in BAL or lung samples.…”

Quantitative proteomic analysis of bile in extrahepatic cholangiocarcinoma patients

“…Another study of bile proteomic analysis that distinguished CCA from PCS showed that ITIH4 was elevated in CCA [9], as is similar to our study but it was not significant. ITIH4 is a serum glycoprotein secreted mainly by the liver [38][39][40], and involved in stabilization of the extracellular matrix [40,41]. A previous study showed that ITIH4 plays a significant role in the pathway involved in TGF-β and fibrosis-independent carcinogenesis [38,42].…”

“…ITIH4 is a serum glycoprotein secreted mainly by the liver [38][39][40], and involved in stabilization of the extracellular matrix [40,41]. A previous study showed that ITIH4 plays a significant role in the pathway involved in TGF-β and fibrosis-independent carcinogenesis [38,42]. In a study of pancreatic cancer biomarkers using pancreatic juice samples, potential biomarkers complement C5 and ITIH3 were elevated in cancer, but biliary obstruction had a significant effect on the performance of the markers [42].…”

Identification of Key Pathways and Genes in Polycystic Ovary Syndrome via Integrated Bioinformatics Analysis and Prediction of Small Therapeutic Molecules

“…F13A1 is an essential regulatory factor to be associated in PCOS development [34]. An investigation has reported that the ITIH4 can promote non-alcoholic fatty liver disease [35], but this gene might be important for progression of PCOS. Da et al [36] have suggested that the TET3 has an important role in controlling type 2 diabetes progression, but this gene might be key role in PCOS.…”