Identification of Significant Potential Signaling Pathways and Differentially Expressed Proteins in Patients with Wheat Intolerance Based on Quantitative Proteomics

Abstract

Background: Wheat intolerance has various systemic manifestations that can affect people’s quality of life, and few studies have focused on the mechanism of wheat intolerance and the signaling pathways involved in wheat intolerance have not been fully identified. Methods: We compared the protein profiles of patients with wheat intolerance with those of healthy controls using LASSO (least absolute shrinkage and selection operator) and PLS (partial least squares regression) to obtain DEPs for Gene Ontology enrichment analysis, Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis, and protein–protein interaction network analysis. Internal validation and external validation were conducted for target proteomics testing. The correlation between differently expressed protein and the wheat-specific IgG antibody concentration was analyzed. Then receiver operating characteristic (ROC) curves was generated to validate the differentially expressed proteins. Findings: We identified 30 DEPs as significant candidate proteins of wheat intolerance. These proteins were enriched in complement and coagulation cascade pathways, regulated exocytosis, immune activation, and immune response-related pathways. After internal and external target proteomics validation, CFHR3 (complement factor H-related protein 3) was identified as a key protein that may have an important role in wheat intolerance. We found CFHR3 protein expression abundance and the wheat-specific IgG antibody concentration were significantly negatively correlated ( P = 0.035; Spearman correlation coefficient r = −0.565). The median area under the ROC curve (AUC) of CFHR3 is 0.857 in external verification data. Interpretation: This study provides insights into wheat intolerance that can be used to further explore the pathogenesis of this condition. Funding Statement: This study was funded by the National Science and Technology Major Project of the Ministry of Science and Technology of China (2016ZX08011005), the National Natural Science Foundation of China (82073666), and the Youth Science Foundation Project of National Natural Science Foundation of China (82003556). Declaration of Interests: None declared. Ethics Approval Statement: This study was approved by Harbin Medical University’s Ethical Review Committee.