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Novel urine cell-free DNA methylation markers for hepatocellular carcinoma
Journal article   Open access   Peer reviewed

Novel urine cell-free DNA methylation markers for hepatocellular carcinoma

Selena Y Lin, Wei Xia, Amy K Kim, Dion Chen, Shelby Schleyer, Lin Choi, Zhili Wang, James P Hamilton, Harry Luu, Hie-Won Hann, …
Scientific reports, v 13(1), pp 21585-21585
07 Dec 2023
PMID: 38062093
url
https://doi.org/10.1038/s41598-023-48500-yView
Published, Version of Record (VoR) Open

Abstract

alpha-Fetoproteins - genetics Biomarkers, Tumor - genetics Carcinoma, Hepatocellular - diagnosis Carcinoma, Hepatocellular - genetics Carcinoma, Hepatocellular - pathology Cell-Free Nucleic Acids - genetics DNA Methylation Humans Liver Neoplasms - diagnosis Liver Neoplasms - genetics Liver Neoplasms - pathology
An optimized hepatocellular carcinoma (HCC)-targeted methylation next generation sequencing assay was developed to discover HCC-associated methylation markers directly from urine for HCC screening. Urine cell-free DNA (ucfDNA) isolated from a discovery cohort of 31 non-HCC and 30 HCC was used for biomarker discovery, identifying 29 genes with differentially methylated regions (DMRs). Methylation-specific qPCR (MSqPCR) assays were developed to verify the selected DMRs corresponding to 8 genes (GRASP, CCND2, HOXA9, BMP4, VIM, EMX1, SFRP1, and ECE). Using archived ucfDNA, methylation of GRASP, HOXA9, BMP4, and ECE1, were found to be significantly different (p < 0.05) between HCC and non-HCC patients. The four markers together with previously reported GSTP1 and RASSF1A markers were assessed as a 6-marker panel in an independent training cohort of 87 non-HCC and 78 HCC using logistic regression modeling. AUROC of 0.908 (95% CI, 0.8656-0.9252) was identified for the 6-marker panel with AFP, which was significantly higher than AFP-alone (AUROC 0.841 (95% CI, 0.778-0.904), p = 0.0026). Applying backward selection method, a 4-marker panel was found to exhibit similar performance to the 6-marker panel with AFP having 80% sensitivity compared to 29.5% by AFP-alone at a specificity of 85%. This study supports the potential use of methylated transrenal ucfDNA for HCC screening.

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Web of Science research areas
Oncology
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