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Global Hypomethylation as Minimal Residual Disease (MRD) Biomarker in Esophageal and Esophagogastric Junction Adenocarcinoma
by
, , , , , , , , , and
Elisa Boldrin
1,*,†,
Maria Assunta Piano
1,†,
Alice Volpato
2,
Rita Alfieri
3,
Monica Franco
4,
Tiziana Morbin
4,
Annalisa Masier
4,
Stefano Realdon
5,
Genny Mattara
6,
Giovanna Magni
7,
Antonio Rosato
1,8,
Pierluigi Pilati
6,
Alberto Fantin
4 and
Matteo Curtarello
1 1
Immunology and Molecular Oncology Diagnostics Unit, Veneto Institute of Oncology IOV-IRCCS, 35128 Padua, Italy
2
Anatomy and Pathological Histology Unit, Veneto Institute of Oncology IOV-IRCCS, 35128 Padua, Italy
3
General Gastric and Esophagus Surgery Unit, IRCCS Humanitas Research Hospital, 20089 Milan, Italy
4
Gastroenterology Unit, Veneto Institute of Oncology IOV-IRCCS, 35128 Padua, Italy
5
Oncological Gastroenterology, Centro di Riferimento Oncologico di Aviano (CRO) IRCCS, 33081 Aviano, Italy
6
Surgical Oncology of Digestive Tract Unit, Veneto Institute of Oncology IOV-IRCCS, 35128 Padua, Italy
7
Clinical Research Unit, Veneto Institute of Oncology IOV-IRCCS, 35128 Padua, Italy
8
Department of Surgery Oncology and Gastroenterology, University of Padova, 35122 Padua, Italy
*
Author to whom correspondence should be addressed.
†
These authors are equal first authors in this work.
Cancers 2025, 17(16), 2668; https://doi.org/10.3390/cancers17162668
Submission received: 28 April 2025
/
Revised: 23 July 2025
/
Accepted: 28 July 2025
/
Published: 15 August 2025
(This article belongs to the Special Issue Circulating Tumour DNA and Liquid Biopsy in Oncology)
Simple Summary
Growing evidence supports the hypothesis that, besides genetic alterations, epigenetic events, in particular aberrant methylation, can also contribute to tumor development and malignant progression that may lead Barrett’s esophagus to evolve into dysplasia and finally into esophageal and esophagogastric junction adenocarcinoma. Moreover, a high relapse frequency after surgery among patients with locally advanced esophageal and esophagogastric junction adenocarcinoma has been observed, suggesting a need to find new biomarkers able to predict the behavior of the disease. Due to these reasons, this research aims to clarify how and when global methylation level is affected during the carcinogenesis process from Barrett’s to adenocarcinoma and if this biomarker could predict relapse. Finding such a biomarker could be of great interest; indeed, it has the potential utility to enable early intervention or adjustment to treatment strategies.
Abstract
Background/Objectives: Esophageal and esophagogastric junction adenocarcinoma (EADC-EGJA), which mainly develops from Barrett’s esophagus (BE), low-grade dysplasia (LGD), and high-grade dysplasia (HGD), has a poor prognosis and several unmet clinical needs, among which is the detection of minimal residual disease (MRD) after endoscopic/surgical resection. Long interspersed nuclear element-1 (LINE-1), a surrogate marker of global methylation, is considered an emerging biomarker for MRD monitoring. The aim of this study was to determine, by LINE-1 methylation analysis, at which carcinogenesis step global methylation is affected and whether this biomarker could be followed in longitudinal to monitor the disease behavior post-surgery. Methods: Cell-free DNA of 90 patients with non-dysplastic Barrett’s esophagus (NDBE), HGD/early EADC-EGJA, or locally advanced/advanced EADC-EGJA were analyzed for LINE-1 methylation, by Methylation-Sensitive Restriction Enzyme droplet digital PCR (MSRE-ddPCR). Twenty-six patients were longitudinally studied by repetitive blood sampling. Results: Global hypomethylation increased during carcinogenesis, with significant difference between locally advanced/advanced EADC-EGJA and NDBE patients (p = 0.028). Longitudinal cases confirmed the rareness of hypomethylation in NDBE cases. The majority of HGD/early EADC-EGJA and locally advanced/advanced EADC-EGJA patients showed methylation changes after resection according to clinical status. Conclusions: This study suggests that global hypomethylation occurs just prior to cancer invasiveness and that it is a promising biomarker to monitor MRD.
