Identification of novel biomarkers of hepatocellular carcinoma by high-definition mass spectrometry: Ultrahigh-performance liquid chromatography quadrupole time-of-flight mass spectrometry and desorption electrospray ionization mass spectrometry imaging.

Koshi Nagai,Daisuke Saigusa,Yoshihisa Tomioka,Shu‐Ping Hui,Hitoshi Ikeda,Zhen Chen,Ritsumi Saito,Amane Fujioka,Hiroki Tsukamoto,Takahiro Yamazaki,Baasanjav Uranbileg,Junken Aoki,Toru Nakazawa,Seizo Koshiba,Yutaka Yatomi,Yotaro Matsumoto,Hitoshi Chiba

doi:10.1002/rcm.8551

Abstract

RationaleHepatocellular carcinoma (HCC) is a highly malignant disease for which the development of prospective or prognostic biomarkers is urgently required. Although metabolomics is widely used for biomarker discovery, there are some bottlenecks regarding the comprehensiveness of detected features, reproducibility of methods, and identification of metabolites. In addition, information on localization of metabolites in tumor tissue is needed for functional analysis. Here, we developed a wide‐polarity global metabolomics (G‐Met) method, identified HCC biomarkers in human liver samples by high‐definition mass spectrometry (HDMS), and demonstrated localization in cryosections using desorption electrospray ionization MS imaging (DESI‐MSI) analysis.MethodsMetabolic profiling of tumor (n = 38) and nontumor (n = 72) regions in human livers of HCC was performed by an ultrahigh‐performance liquid chromatography quadrupole time‐of‐flight MS (UHPLC/QTOFMS) instrument equipped with a mixed‐mode column. The HCC biomarker candidates were extracted by multivariate analyses and identified by matching values of the collision cross section and their fragment ions on the mass spectra obtained by HDMS. Cryosections of HCC livers, which included both tumor and nontumor regions, were analyzed by DESI‐MSI.ResultsFrom the multivariate analysis, m/z 904.83 and m/z 874.79 were significantly high and low, respectively, in tumor samples and were identified as triglyceride (TG) 16:0/18:1(9Z)/20:1(11Z) and TG 16:0/18:1(9Z)/18:2(9Z,12Z) using the synthetic compounds. The TGs were clearly localized in the tumor or nontumor areas of the cryosection.ConclusionsNovel biomarkers for HCC were identified by a comprehensive and reproducible G‐Met method with HDMS using a mixed‐mode column. The combination analysis of UHPLC/QTOFMS and DESI‐MSI revealed that the different molecular species of TGs were associated with tumor distribution and were useful for characterizing the progression of tumor cells and discovering prospective biomarkers.

Highlights

Hepatocellular carcinoma (HCC), the primary type of liver cancer, is highly malignant and is continuing to increase worldwide
In the study reported here, we developed a wide‐polarity global metabolomics/metabolic profiling (G‐Met) method using a mixed‐mode column and identified biomarker candidates for HCC in human liver samples using High‐definition mass spectrometry (HDMS)
We first evaluated the utility of G‐Met by UHPLC/QTOFMS with a mixed‐mode column for the analysis of multiple biological liver samples

Summary

Introduction

Hepatocellular carcinoma (HCC), the primary type of liver cancer, is highly malignant and is continuing to increase worldwide. In the study reported here, we developed a wide‐polarity G‐Met method using a mixed‐mode column and identified biomarker candidates for HCC in human liver samples using HDMS.

Results

Conclusion