Background: Colorectal cancer (CRC) can be cured when diagnosed in its early or precancerous
(adenoma) stages. Mostly due to poor compliance towards invasive screening procedures,
detection rates for adenoma and early CRCs are still low. Available non-invasive screening
tests have unfortunately low sensitivity and specificity performances. Therefore, there is a
large unmet need calling for a cost-effective, reliable and non-invasive test to screen for
early neoplastic and pre-neoplastic lesions. Objective: To develop a routine screening test
based on a nucleic acids multi-gene assay performed on peripheral blood mononuclear cells
(PBMCs) that can detect early CRCs and adenomas. Methods: 116 patients (mean age: 55
years; range: 18 to 74 years; female/male ration 0.98) were included in this pilot, nonblinded,
colonoscopy-controlled study. Colonoscopy revealed 21 patients with CRC, 30
patients with adenoma bigger than 1 cm, 24 patients with inflammatory bowel disease (IBD)
and 41 patients had no neoplastic or inflammatory lesions. Blood samples were taken from
each patient the day of the colonoscopy and PBMCs were purified. Total RNA was extracted
following standard procedures. Multiplex RT-qPCR was applied on 92 different candidate
biomarkers. Different univariate and multivariate statistical methods were applied on these
candidates, and among them, 57 biomarkers with significant p values (<0.01, Wilcoxon
test) were selected, including ADAMTS1, MMP9, CXCL10, CXCR4, VEGFA and CDH1.
Two distinct biomarker signatures are used to separate patients without neoplastic lesion
from those with cancer (named COLOX 1 test), respectively from those with adenoma
(named COLOX 2 test). Result: COLOX 1 and 2 tests have successfully separated patients
without neoplastic lesion from those with CRC (sensitivity 70%, specificity 90%, AUC 0.88),
respectively from those with adenoma bigger than 1cm (sensitivity 61%, specificity 80%,
AUC 0.80). 6/24 patients in the IBD group have a positive COLOX 1 test. Conclusion:
These two COLOX tests demonstrated an acceptable sensitivity and a high specificity to
detect the presence of CRCs and adenomas bigger than 1 cm. The false positives COLOX
1 test in IBD patients could possibly be due to the chronic inflammatory state. A prospective,
multicenter, pivotal study is underway in order to confirm these promising results in a
larger cohort.