Non-invasive colorectal cancer biomarkers: HAND2 and GPM6A methylation in circulating tumour DNA

Abstract Background Colorectal cancer (CRC) is a leading cause of cancer-related deaths worldwide. Early detection is critical for improving patient outcomes. This study investigated the potential of circulating tumour DNA (ctDNA) methylation markers, specifically Heart and Neural Crest Derivatives...

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Main Authors: Amir Mahdi Nili, Kamal Mohammadian, Fatemeh Namazi Nanehkaran, Faranak Jamshidian, Arash Moradi, Shahla Mohammad Ganji
Format: Article
Language:English
Published: BMC 2025-07-01
Series:Cancer Cell International
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Online Access:https://doi.org/10.1186/s12935-025-03898-5
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author Amir Mahdi Nili
Kamal Mohammadian
Fatemeh Namazi Nanehkaran
Faranak Jamshidian
Arash Moradi
Shahla Mohammad Ganji
author_facet Amir Mahdi Nili
Kamal Mohammadian
Fatemeh Namazi Nanehkaran
Faranak Jamshidian
Arash Moradi
Shahla Mohammad Ganji
author_sort Amir Mahdi Nili
collection DOAJ
description Abstract Background Colorectal cancer (CRC) is a leading cause of cancer-related deaths worldwide. Early detection is critical for improving patient outcomes. This study investigated the potential of circulating tumour DNA (ctDNA) methylation markers, specifically Heart and Neural Crest Derivatives Expressed 2 (HAND2) and Glycoprotein M6A (GPM6A), as novel non-invasive biomarkers for CRC detection and treatment monitoring. Methods Plasma samples were collected from 40 CRC patients before and after treatment, as well as 40 healthy controls. Methylation levels of HAND2 and GPM6A in ctDNA were quantified using quantitative methylation-specific PCR (qMS-PCR). Results Methylation levels of HAND2 and GPM6A were significantly elevated in pre-treatment colorectal cancer (CRC) patients compared to post-treatment patients and healthy controls. Specifically, HAND2 methylation exhibited a statistically significant reduction following therapeutic intervention (pre-treatment: 55.75% ± 26.75 vs. post-treatment: 37.98% ± 22.93, p < 0.001), with no significant correlation observed between pre- and post-treatment levels (Pearson r = 0.1030, p = 0.53). Similarly, GPM6A methylation decreased markedly post-treatment (pre-treatment: 72.93% ± 18.06 vs. post-treatment: 49.48% ± 16.83, p < 0.001) yet demonstrated a statistically significant positive correlation between pre- and post-treatment values (Pearson r = 0.3181, p = 0.05). Receiver operating characteristic (ROC) curve analysis revealed robust diagnostic performance, with the combined HAND2 and GPM6A methylation model yielding an area under the curve (AUC) of 0.9213 (95% CI: 0.8569–0.9856, p < 0.001), surpassing the individual discriminative capacities of HAND2 (AUC = 0.8906) and GPM6A (AUC = 0.8881). Survival analysis leveraging the Cancer Genome Atlas (TCGA) dataset indicated a potential association between elevated methylation of both genes. It reduced survival probability, with a pronounced effect observed in rectal adenocarcinoma. Conclusion HAND2 and GPM6A methylation in ctDNA are promising non-invasive biomarkers for CRC detection and monitoring, outperforming the diagnostic accuracy of traditional markers such as CEA or CA19-9. Further research is needed to validate these findings in larger cohorts and investigate their potential as markers for treatment response and disease recurrence.
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spelling doaj-art-7e7e37da06484aefa496410f85a027b92025-08-20T03:43:36ZengBMCCancer Cell International1475-28672025-07-0125111110.1186/s12935-025-03898-5Non-invasive colorectal cancer biomarkers: HAND2 and GPM6A methylation in circulating tumour DNAAmir Mahdi Nili0Kamal Mohammadian1Fatemeh Namazi Nanehkaran2Faranak Jamshidian3Arash Moradi4Shahla Mohammad Ganji5Department of Biology, Tehran East Branch, Basic Sciences, Islamic Azad UniversityDepartment of Radiation Oncology, Faculty of Medicine, Hamadan University of Medical SciencesDepartment of Radiation Oncology, Faculty of Medicine, Hamadan University of Medical SciencesDepartment of Biology, Tehran East Branch, Basic Sciences, Islamic Azad UniversityDepartment of Molecular Medicine, Medical Biotechnology Institute, National Institute of Genetic Engineering and Biotechnology (NIGEB)Department of Molecular Medicine, Medical Biotechnology Institute, National Institute of Genetic Engineering and Biotechnology (NIGEB)Abstract Background Colorectal cancer (CRC) is a leading cause of cancer-related deaths worldwide. Early detection is critical for improving patient outcomes. This study investigated the potential of circulating tumour DNA (ctDNA) methylation markers, specifically Heart and Neural Crest Derivatives Expressed 2 (HAND2) and Glycoprotein M6A (GPM6A), as novel non-invasive biomarkers for CRC detection and treatment monitoring. Methods Plasma samples were collected from 40 CRC patients before and after treatment, as well as 40 healthy controls. Methylation levels of HAND2 and GPM6A in ctDNA were quantified using quantitative methylation-specific PCR (qMS-PCR). Results Methylation levels of HAND2 and GPM6A were significantly elevated in pre-treatment colorectal cancer (CRC) patients compared to post-treatment patients and healthy controls. Specifically, HAND2 methylation exhibited a statistically significant reduction following therapeutic intervention (pre-treatment: 55.75% ± 26.75 vs. post-treatment: 37.98% ± 22.93, p < 0.001), with no significant correlation observed between pre- and post-treatment levels (Pearson r = 0.1030, p = 0.53). Similarly, GPM6A methylation decreased markedly post-treatment (pre-treatment: 72.93% ± 18.06 vs. post-treatment: 49.48% ± 16.83, p < 0.001) yet demonstrated a statistically significant positive correlation between pre- and post-treatment values (Pearson r = 0.3181, p = 0.05). Receiver operating characteristic (ROC) curve analysis revealed robust diagnostic performance, with the combined HAND2 and GPM6A methylation model yielding an area under the curve (AUC) of 0.9213 (95% CI: 0.8569–0.9856, p < 0.001), surpassing the individual discriminative capacities of HAND2 (AUC = 0.8906) and GPM6A (AUC = 0.8881). Survival analysis leveraging the Cancer Genome Atlas (TCGA) dataset indicated a potential association between elevated methylation of both genes. It reduced survival probability, with a pronounced effect observed in rectal adenocarcinoma. Conclusion HAND2 and GPM6A methylation in ctDNA are promising non-invasive biomarkers for CRC detection and monitoring, outperforming the diagnostic accuracy of traditional markers such as CEA or CA19-9. Further research is needed to validate these findings in larger cohorts and investigate their potential as markers for treatment response and disease recurrence.https://doi.org/10.1186/s12935-025-03898-5Colorectal cancerNon-invasive biomarkerCtDNAGPM6AHAND2
spellingShingle Amir Mahdi Nili
Kamal Mohammadian
Fatemeh Namazi Nanehkaran
Faranak Jamshidian
Arash Moradi
Shahla Mohammad Ganji
Non-invasive colorectal cancer biomarkers: HAND2 and GPM6A methylation in circulating tumour DNA
Cancer Cell International
Colorectal cancer
Non-invasive biomarker
CtDNA
GPM6A
HAND2
title Non-invasive colorectal cancer biomarkers: HAND2 and GPM6A methylation in circulating tumour DNA
title_full Non-invasive colorectal cancer biomarkers: HAND2 and GPM6A methylation in circulating tumour DNA
title_fullStr Non-invasive colorectal cancer biomarkers: HAND2 and GPM6A methylation in circulating tumour DNA
title_full_unstemmed Non-invasive colorectal cancer biomarkers: HAND2 and GPM6A methylation in circulating tumour DNA
title_short Non-invasive colorectal cancer biomarkers: HAND2 and GPM6A methylation in circulating tumour DNA
title_sort non invasive colorectal cancer biomarkers hand2 and gpm6a methylation in circulating tumour dna
topic Colorectal cancer
Non-invasive biomarker
CtDNA
GPM6A
HAND2
url https://doi.org/10.1186/s12935-025-03898-5
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