AUTHOR=Zhang Hao , Zuo Lugen , Li Jing , Geng Zhijun , Ge Sitang , Song Xue , Wang Yueyue , Zhang Xiaofeng , Wang Lian , Zhao Tianhao , Deng Min , Chai Damin , Wang Qiusheng , Yang Zi , Liu Quanli , Qiu Quanwei , He Xuxu , Yang Yiqun , Ge Yuanyuan , Wu Rong , Zheng Lin , Li Jianjun , Chen Runkai , Sun Jialiang , Hu Jianguo 

TITLE=Construction of a fecal immune-related protein-based biomarker panel for colorectal cancer diagnosis: a multicenter study

JOURNAL=Frontiers in Immunology

VOLUME=Volume 14 - 2023

YEAR=2023

URL=https://www.frontiersin.org/journals/immunology/articles/10.3389/fimmu.2023.1126217

DOI=10.3389/fimmu.2023.1126217

ISSN=1664-3224

ABSTRACT=Purpose: To explore fecal immune-related proteins that can be used for colorectal cancer (CRC) diagnosis.
Patients and Methods:
Three independent cohorts were used in present study. In the discovery cohort, which included 14 CRC patients and 6 healthy controls (HCs), label-free proteomics was applied to identify immune-related proteins in stool that could be used for CRC diagnosis. Exploring potential links between gut microbes and immune-related proteins by 16S rRNA sequencing. The abundance of fecal immune-associated proteins was verified by ELISA in two independent validation cohorts and a biomarker panel was constructed that could be used for CRC diagnosis. The validation cohort I included 192 CRC patients and 151 HCs from 6 different hospitals. The validation cohort II included 141 CRC patients, 82 colorectal adenoma (CRA) patients, and 87 HCs from another hospital. Finally, the expression of biomarkers in cancer tissues was verified by immunohistochemistry (IHC).
Results:
In the discovery study, 436 plausible fecal proteins were identified. And among 67 differential fecal proteins (|log2 fold change| > 1, P < 0.01) that could be used for CRC diagnosis, 16 immune-related proteins with diagnostic value were identified. The 16S rRNA sequencing results showed a positive correlation between immune-related proteins and the abundance of oncogenic bacteria. In the first validation cohort, a biomarker panel consisting of five fecal immune-related proteins (CAT, LTF, MMP9, RBP4, and SERPINA3) was constructed based on the least absolute shrinkage and selection operator (LASSO) and multivariate logistic regression. In two independent validation cohorts, this biomarker panel was superior to hemoglobin in the diagnosis of CRC. The IHC result showed that protein expression levels of these five immune-related proteins were significantly higher in CRC tissue than in normal colorectal tissue.
Conclusion: A novel biomarker panel consisting of fecal immune-related proteins can be used for the diagnosis of CRC.