Detection of Colorectal Cancer in Circulating Cell-Free DNA by Methylated CpG Tandem Amplification and Sequencing

Clin Chem. 2019 Jul;65(7):916-926. doi: 10.1373/clinchem.2019.301804. Epub 2019 Apr 22.

Abstract

Background: Aberrant DNA hypermethylation of CpG islands occurs frequently throughout the genome in human colorectal cancer (CRC). A genome-wide DNA hypermethylation analysis technique using circulating cell-free DNA (cfDNA) is attractive for the noninvasive early detection of CRC and discrimination between CRC and other cancer types.

Methods: We applied the methylated CpG tandem amplification and sequencing (MCTA-Seq) method, with a fully methylated molecules algorithm, to plasma samples from patients with CRC (n = 147) and controls (n = 136), as well as cancer and adjacent noncancerous tissue samples (n = 66). We also comparatively analyzed plasma samples from patients with hepatocellular carcinoma (HCC; n = 36).

Results: Dozens of DNA hypermethylation markers including known (e.g., SEPT9 and IKZF1) and novel (e.g., EMBP1, KCNQ5, CHST11, APBB1IP, and TJP2) genes were identified for effectively detecting CRC in cfDNA. A panel of 80 markers discriminated early-stage CRC patients and controls with a clinical sensitivity of 74% and clinical specificity of 90%. Patients with early-stage CRC and HCC could be discriminated at clinical sensitivities of approximately 70% by another panel of 128 markers.

Conclusions: MCTA-Seq is a promising method for the noninvasive detection of CRC.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aged
  • Algorithms
  • Base Sequence
  • Biomarkers, Tumor / blood*
  • Biomarkers, Tumor / genetics
  • Carcinoma, Hepatocellular / diagnosis
  • Cell-Free Nucleic Acids / blood*
  • Cell-Free Nucleic Acids / genetics
  • Colorectal Neoplasms / diagnosis*
  • CpG Islands*
  • DNA Methylation
  • Diagnosis, Differential
  • Female
  • Humans
  • Liver Neoplasms / diagnosis
  • Male
  • Middle Aged
  • Nucleic Acid Amplification Techniques / methods
  • Principal Component Analysis

Substances

  • Biomarkers, Tumor
  • Cell-Free Nucleic Acids