Evaluation and Validation of Plasma Proteins Using Two Different Protein Detection Methods for Early Detection of Colorectal Cancer.

Christoph H. Borchers,Axel Benner,Anton Gies,Petra Schrotz-King,Hermann Brenner,Korbinian Weigl,Kaja Tikk,Megha Bhardwaj

doi:10.3390/cancers11101426

Abstract

Objective: Plasma protein biomarkers could be an efficient alternative for population-based screening for early detection of colorectal cancer (CRC). The objective of this study was to evaluate and validate plasma proteins individually and as a signature for early detection of CRC. Methods: In a three-stage design, proteins were measured firstly by liquid chromatography/multiple reaction monitoring-mass spectrometry (LC/MRM-MS) and later by proximity extension assay (PEA) in a discovery set consisting of 96 newly diagnosed CRC cases and 94 controls free of neoplasms at screening colonoscopy. Two algorithms (one for each measurement method) were derived by Lasso regression and .632+ bootstrap based on 11 proteins that were included in both the LC/MRM-MS and PEA measurements. Additionally, another algorithm was constructed from the same eleven biomarkers plus amphireglin, the most promising protein marker in the PEA measurements that had not been available from the LC/MRM-MS measurements. Lastly the three prediction signatures were validated with PEA in independent samples of participants of screening colonoscopy (CRC (n = 56), advanced adenoma (n = 101), and participants free of neoplasm (n = 102)). Results: The same four proteins were included in all three prediction signatures; mannan binding lectin serine protease 1, osteopontin, serum paraoxonase lactonase 3 and transferrin receptor protein 1, and the third prediction signature additionally included amphiregulin. In the independent validation set from a true screening setting, the five-marker blood-based signature including AREG presented areas under the curves of 0.82 (95% CI, 0.74–0.89), 0.86 (95% CI, 0.77–0.92) and 0.76 (95% CI, 0.64–0.86) for all, early and late stages CRC, respectively. Conclusion: Two different measurement methods consistently identified four protein markers and an algorithm additionally including amphiregulin, a marker measured by PEA only, showed promising performance for detecting early stage CRC in an independent validation in a true screening setting. These proteins may be potential candidates for blood-based tests for early detection of CRC.

Highlights

Colorectal cancer (CRC) contributes immensely to the global burden of cancers with 1.85 million incident cases and approximately 880,000 deaths per year
When the estimates were validated independently on participants of screening colonoscopy for all stage CRC detection AUCs of 0.76 and 0.75 were observed for the LC/MRM-MS and proximity extension assay (PEA) algorithms from the discovery set, respectively
The four-marker signature yielded AUCs of 0.78 and 0.75 for early stage CRC detection and AUCs of 0.71 and 0.72 for late stage CRC detection, for LC/MRM-MS and PEA algorithms, respectively

Summary

Introduction

Colorectal cancer (CRC) contributes immensely to the global burden of cancers with 1.85 million incident cases and approximately 880,000 deaths per year. CRC is the third most common cancer and second leading cause of cancer mortality globally [1]. There is increasing evidence that CRC incidence and mortality could be effectively reduced by screening [2,3,4,5]. The participation rates in screening programs are often low due to inconvenience and invasiveness in endoscopy-based programs [6,7] and by reservation against collection, handling and storage of stool in stool test based screening programs [8]. The participation in population-based screening programs could potentially improve with minimally invasive blood-based tests [9]. Multiplex platforms like liquid chromatography/multiple reaction monitoring-mass spectrometry (LC/MRM-MS) [11,12] and proximity extension assay (PEA) [13,14]

Methods

Results

Discussion

Conclusion