Early detection of colorectal cancer based on presence of methylated syndecan-2 (SDC2) in stool DNA

Yoon Dae Han, Tae Jeong Oh, Tae-Ha Chung, Hui Won Jang, Youn Nam Kim, Sungwhan An, Nam Kyu Kim, Yoon Dae Han, Tae Jeong Oh, Tae-Ha Chung, Hui Won Jang, Youn Nam Kim, Sungwhan An, Nam Kyu Kim

Abstract

Background: Colorectal cancer (CRC) screening can effectively reduce disease-related mortality by detecting CRC at earlier stages. We have previously demonstrated that the presence of SDC2 methylation in stool DNA is significantly associated with the occurrence of CRC regardless of clinical stage. The aim of this study was to evaluate the clinical performance of stool DNA-based SDC2 methylation test for CRC.

Methods: Aberrant SDC2 methylation in stool-derived DNA was measured by linear target enrichment (LTE)-quantitative methylation-specific real-time PCR (qMSP). Duplicate reactions of meSDC2 LTE-qMSP test were performed for stool samples obtained from CRC patients representing all stages (0-IV) and asymptomatic individuals who were subsequently underwent colonoscopy examination. To determine the diagnostic value of test in CRC and control groups, sensitivity and specificity were evaluated by receiver operating characteristic curve analysis.

Results: Of 585 subjects who could be evaluated, 245 had CRC, 44 had various sizes of adenomatous polyps, and 245 had negative colonoscopy results. Stool DNA-based meSDC2 LTE-qMSP showed an overall sensitivity of 90.2% with AUC of 0.902 in detecting CRC (0-IV) not associated with tumor stage, location, sex, or age (P > 0.05), with a specificity of 90.2%. Sensitivity for detecting early stages (0-II) was 89.1% (114/128). This test also detected 66.7% (2/3) and 24.4% (10/41) of advanced and non-advanced adenomas, respectively.

Conclusions: Results of this study validated the capability of stool DNA based-SDC2 methylation test by LTE-qMSP for early detection of CRC patient with high specificity.

Trial registration: ClinicalTrials.gov, NCT03146520 , Registered 10 May 2017, Retrospectively registered; however, control arm was prospectively registered.

Keywords: Biomarker; Colorectal cancer; DNA methylation; Early detection; Syndecan-2.

Conflict of interest statement

Ethics approval and consent to participate

This study was approved by the Institutional Review Board of College of Medicine of Yonsei University College of Medicine (IRB No. 1–2016-0068).

Consent for publication

Not applicable.

Competing interests

TaeJeong Oh and Sungwhan An are employees of Genomictree, Inc. TaeJeong Oh and Sungwhan An are shareholders of Genomictree, Inc. The other authors state no other conflict of interest.

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Figures

Fig. 1
Fig. 1
Study enrollment and flow diagram. The number of enrolled subjects according to diagnosis and the number of stool samples analyzed in this study. a Samples had insufficient DNA, b Samples either had insufficient human genomic DNA (low COL2A1) or unknown clinical stage
Fig. 2
Fig. 2
LoD of SDC2 methylation test using data derived from Table 2. The LoD of the test using simulated stool DNA samples was determined by Probit analysis. The y-axis plots the probability of methylated SDC2 detection while the x-axis plots spiked HCT116 genomic DNA in a total of 2.0 μg of SDC2 methylation-negative stool genomic DNA. The estimated LoD (34.5 pg) is indicated by an arrow
Fig. 3
Fig. 3
Results of SDC2 methylation analysis from two reactions in stool DNA. Distribution of SDC2 methylation was expressed in CT values as 40-CT for each sample. A higher 40-CT represents a higher methylation level of SDC2. It is represented as 0 if the SDC2 methylation was not detectable. Methylation status of SDC2 gene is plotted as box and whisker plots. CRC colorectal cancer (stage 0–IV), HOP hyperplastic or other polyps, NA non-advanced adenomas (< 1.0 cm), AA advanced adenomas (≥ 1.0 cm), GC gastric cancer, LC liver cancer, NED no evidence of disease
Fig. 4
Fig. 4
Results of stool DNA-based SDC2 methylation test. a ROC curve was plotted for CRC patients vs. subjects with NED. AUC is indicated. b Sensitivity of SDC2 methylation test for detecting CRC according to clinical stages. Percent of samples with detectable methylated SDC2 DNA using 1/2 algorithm is depicted by solid bars

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