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

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

doi:10.3389/fimmu.2023.1126217

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

Front Immunol. 2023 May 29:14:1126217. doi: 10.3389/fimmu.2023.1126217. eCollection 2023.

Authors

Hao Zhang¹, Lugen Zuo^{1

2}, Jing Li^{2

3}, Zhijun Geng^{2

4}, Sitang Ge^{1

2}, Xue Song^{2

4}, Yueyue Wang^{2

3}, Xiaofeng Zhang^{2

4}, Lian Wang¹, Tianhao Zhao^{2

5}, Min Deng^{2

5}, Damin Chai⁶, Qiusheng Wang¹, Zi Yang¹, Quanli Liu¹, Quanwei Qiu¹, Xuxu He¹, Yiqun Yang¹, Yuanyuan Ge⁷, Rong Wu⁸, Lin Zheng⁹, Jianjun Li¹⁰, Runkai Chen¹¹, Jialiang Sun¹², Jianguo Hu^{2

3}

Affiliations

¹ Department of Gastrointestinal Surgery, First Affiliated Hospital of Bengbu Medical College, Bengbu, China.
² Department of Inflammatory Bowel Disease Research Center, First Affiliated Hospital of Bengbu Medical College, Bengbu, China.
³ Department of Clinical Laboratory, First Affiliated Hospital of Bengbu Medical College, Bengbu, China.
⁴ Department of Central Laboratory, First Affiliated Hospital of Bengbu Medical College, Bengbu, China.
⁵ Department of Gastroenterology, First Affiliated Hospital of Bengbu Medical College, Bengbu, China.
⁶ Department of Pathology, First Affiliated Hospital of Bengbu Medical College, Bengbu, China.
⁷ Department of Colorectal Surgery, Nanjing Hospital of Chinese Medicine Affiliated to Nanjing University of Chinese Medicine, Nanjing, China.
⁸ Department of General Surgery, Zhongda Hospital, Southeast University, Nanjing, China.
⁹ Department of Clinical Laboratory, The Affiliated Hospital of Medical School of Ningbo University, Ningbo, China.
¹⁰ Department of General Surgery, The First Medical Center, Chinese PLA General Hospital, Beijing, China.
¹¹ Department of General Surgery, Chinese PLA General Hospital, Beijing, China.
¹² Department of General Surgery, Shanghai Fengxian District Central Hospital, Shanghai, China.

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 validation cohort I, 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. The biomarker panel was found to be superior to hemoglobin in the diagnosis of CRC in both validation cohort I and validation cohort II. 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.

Keywords: colorectal cancer; fecal protein; immune-related protein; multicenter study; proteomics.

Publication types

Multicenter Study
Research Support, Non-U.S. Gov't

MeSH terms

Biomarkers
Colorectal Neoplasms* / diagnosis
Enzyme-Linked Immunosorbent Assay
Feces
Humans
RNA, Ribosomal, 16S / genetics
Retinol-Binding Proteins, Plasma

Substances

RNA, Ribosomal, 16S
Biomarkers
RBP4 protein, human
Retinol-Binding Proteins, Plasma

Grants and funding

This work was supported partly by funding from the University Collaborative Innovation Project of Anhui Province (GXXT-2020-020), the National Natural Science Foundation of China (82070561), the Provincial Health Research Project of Anhui (AHWJ2022a019, AHWJ2022b088).