Proteome Multimarker Panel With Multiple Reaction Monitoring–Mass Spectrometry for Early Detection of Hepatocellular Carcinoma

Abstract

There is an urgent need for new biomarkers that address the shortcomings of current screening methods which fail to detect a large proportion of cases with hepatocellular carcinoma (HCC) at early stage. To develop a robust, multiple‐biomarker panel based on multiple reaction monitoring–mass spectrometry with high performance in detecting early‐stage HCC within at‐risk populations. In the discovery set, 150 samples were analyzed to identify candidate biomarkers. The resulting list of candidates was tested in the training set (713 samples) to establish a multimarker panel, which was evaluated in the validation set (305 samples). We identified 385 serum HCC biomarker candidates in the discovery set and developed a multimarker panel consisting of 28 peptides that best differentiated HCC from controls. The area under the receiver operating characteristic curve of multimarker panel was significantly higher than alpha‐fetoprotein (AFP) in the training (0.976 vs. 0.804; P < 0.001) and validation (0.898 vs. 0.778; P < 0.001) sets. In the validation set, this multimarker panel, compared with AFP, showed significantly greater sensitivity (81.1% vs. 26.8%; P < 0.001) and lower specificity (84.8% vs. 98.8%; P < 0.001) in detecting HCC cases. Combining AFP with the multimarker panel did not significantly improve the area under the receiver operating characteristic curve compared with the panel alone in the training (0.981 vs. 0.976; P = 0.37) and validation set (0.906 vs. 0.898; P = 0.75). Conclusion: The multiple reaction monitoring–mass spectrometry multimarker panel consisting of 28 peptides discriminates HCC cases from at‐risk controls with high performance and may have potential for clinical application in HCC surveillance.

Proteome Multimarker Panel for the Early Detection of Hepatocellular Carcinoma: Multicenter Derivation, Validation, and Comparison

“…A subsequent quantitative MRM-MS analysis targeted 385 peptides that had undergone an interference screen using a stable isotope-labeled standard peptide mixture. 17 …”

“… 11 Similarly, multiple reaction monitoring–mass spectrometry (MRM-MS), a targeted proteomic approach, quickly and accurately analyzes hundreds of proteins as potential biomarkers in various types of cancer. 12 − 16 Our group has proposed a multimarker panel for the detection of early-stage HCC, consisting of 28 proteins that have been identified by MRM-MS. 17 …”

“…11 Similarly, multiple reaction monitoring−mass spectrometry (MRM-MS), a targeted proteomic approach, quickly and accurately analyzes hundreds of proteins as potential biomarkers in various types of cancer. 12−16 Our group has proposed a multimarker panel for the detection of early-stage HCC, consisting of 28 proteins that have been identified by MRM-MS. 17 This multicenter study aimed to develop and evaluate a new multimarker panel for the detection of HCC with target proteins using a previous proteomic dataset, examine whether the incorporation of AFP and PIVKA-II into the panel improves its accuracy, validate the panel in an independent cohort, and compare its performance with that of the GALAD score. ■ METHODS Study Design and Participants.…”

“…A subsequent quantitative MRM-MS analysis targeted 385 peptides that had undergone an interference screen using a stable isotope-labeled standard peptide mixture. 17 In this study, we performed MRM-MS quantification for 383 target peptides, correlating to 176 proteins, except for two peptides for which compatible stable isotope-labeled standard peptides were not obtained due to unavailability. Their exclusion did not affect the performance of the developed panel because the two peptides were part of proteins that were represented by other peptides in the MRM-MS quantification.…”

Proteomic Profiling and Artificial Intelligence for Hepatocellular Carcinoma Translational Medicine

“…For example, in a recent study, high performance multiple reaction monitoring mass spectrometry (MRM-MS) was utilized in detecting early-stage HCC within at-risk populations to allow identification of 385 serum HCC biomarker candidates. A multimarker panel consisting of 28 peptides has been created that best differentiated HCC from controls [ 181 ]. This multimarker panel showed significantly greater sensitivity (81.1% vs. 26.8%) and lower specificity (84.8% vs. 98.8%) in detecting HCC cases as compared to AFP.…”