Identification and Validation of Stage‐Associated Serum Biomarkers in Colorectal Cancer Using MS‐Based Procedures

Abstract

Purpose Successful prevention of colorectal cancer (CRC) would benefit from a rapid serum screening for early detection. Here, a novel strategy for CRC biomarker discovery and validation exclusively based on MS procedures is reported. Experimental design Identification of CRC serum biomarkers is initially made using label‐free quantification on pooled serum samples from different CRC stages followed by two consecutive steps of targeted parallel reaction monitoring assays in different serum cohorts. Relevance of different protein depletion and peptide fractionation extent is investigated. Absolute quantification of a selected peptide is performed as a proof‐of‐concept. Results A total of 945 proteins showed differential abundance in the discovery phase. Based on their statistical significance and relative expression in disease stages, 123 potential biomarkers are selected for a training step. In the final validation step, five peptides belonging to four proteins are consistently quantified in individual CRC serum samples and controls. Different statistical analyses indicate that peptides GWVTDGFSSLK (APOC3) and LCNNPTPQFGGK (THBS1) are candidate biomarkers. Absolute quantification of LCNNPTPQFGGK shows statistical significance for the diagnosis of early respect to late CRC stages. Conclusions and clinical relevance Two peptides from APOC3 and THBS1 are validated by PRM as potential biomarkers for non‐invasive diagnosis of colorectal cancer.

Clinical Assay for the Early Detection of Colorectal Cancer Using Mass Spectrometric Wheat Germ Agglutinin Multiple Reaction Monitoring

“…Further, many studies have applied MS with machine learning algorithms to develop diagnostic tools in CRC diagnosis. Marin-Vincente et al utilized THBS1 and APOC3 with a decision tree classifier and achieved an AUC of 0.83 (with a sensitivity of 90% and a specificity of 65%) in CRC diagnoses [ 51 ]. Another study carried out by Xie et al used CELA1, CEL2A, CTRL, and TRY2 with a logistic regression and achieved an AUC of 0.90 (with a sensitivity of 86.7% and a specificity of 83.3%) in CRC diagnoses [ 52 ].…”

An Aging-Related Gene Signature-Based Model for Risk Stratification and Prognosis Prediction in Lung Adenocarcinoma

“…The majority of ARGs included in our signature are closely related to tumor initiation, proliferation, and metastasis. Apolipoprotein C3 (APOC3) is primarily present on the surface of triglyceride-rich lipoproteins and has been reported as an independent contributor to triglyceride levels (Marín-Vicente et al, 2020). APOC3 is a potential prognostic biomarker for several types of malignant tumors Marín-Vicente et al, 2020).…”

“…Apolipoprotein C3 (APOC3) is primarily present on the surface of triglyceride-rich lipoproteins and has been reported as an independent contributor to triglyceride levels (Marín-Vicente et al, 2020). APOC3 is a potential prognostic biomarker for several types of malignant tumors Marín-Vicente et al, 2020). Erythropoietin receptor (EPOR), a glycoprotein produced in the fetal liver and adult kidney, is the chief regulator of erythropoiesis (Fisher, 2003;Oshima et al, 2021).…”

Proteomic Analyses of Fibroblast- and Serum-Derived Exosomes Identify QSOX1 as a Marker for Non-invasive Detection of Colorectal Cancer

“…Regulatory effects on angiogenesis have been shown for FBLN1 [ 63 ], QSOX1 [ 78 ], THBS1 [ 79 , 80 ] and EDIL3 [ 69 ]. In addition, an association with fibroblasts/CAFs [ 80 , 81 , 82 , 83 , 84 , 85 , 86 , 87 ] and EVs [ 64 , 66 , 72 , 76 , 88 , 89 , 90 , 91 , 92 ] was described for each protein. In tumour tissue, the expression of all six markers was shown to be of prognostic value in several tumour types [ 71 , 72 , 77 , 93 , 94 , 95 ].…”