Cell-free DNA methylation profile potential in the diagnosis of lung squamous cell carcinoma

Q: What was the main goal of the study?

To evaluate whether methylation patterns in lung squamous cell carcinoma (LUSC) and matched non-tumor samples can support LUSC diagnosis, including the potential of cell-free DNA (cfDNA) methylation analysis.

Q: How were methylation profiles generated and compared?

Tumor tissue, peritumoral tissue, distant normal tissue, and corresponding plasma samples were collected from LUSC patients and profiled using a bespoke lung cancer bisulfite sequencing panel; control plasma came from healthy volunteers.

Q: What diagnostic value did the researchers observe?

A cfDNA-based diagnostic panel using a combined diagnostic score (cd-score) showed high sensitivity and specificity, and LUSC patients had significantly higher cd-scores than healthy controls.

Sep 21, 2026

Uploaded by Anda

Aberrant DNA methylation plays a critical role in early cancer development, motivating an investigation of methylation patterns in lung squamous cell carcinoma (LUSC) across tumor, matched non-tumor tissues, and plasma. The study analyzed 49 stage I–III LUSC patients using a lung cancer bisulfite sequencing panel and 48 healthy volunteers for control plasma. Tumor and surrounding tissues showed distinct methylation signatures, enabling identification of tumor-specific differentially methylated blocks and classification into two methylation subgroups with differing diagnostic scores. Cell-free DNA methylation analysis demonstrated potential clinical utility for LUSC diagnosis.

LanguageEnglish

CategoryResearch & Report

FormatPDF

Pages11

Reading Time28 min

AccessFree to download

Background

Methods

Results

Conclusions

Introduction

Cell-free DNA methylation profile potential in the diagnosis of lungsquamous cell carcinoma Chengli Du1, Jin Cai2, Jie Tang1, Yunhao Chen1, Roberto Díaz-Peña3,4, Yusuke Tomita5, Jacek Jassem6, Jiangang Zhao1, Difang Zheng1, Zhengliang Tu1 1Department of Thoracic Surgery, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China; 2Special Clinical Lab, The Third Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou, China; 3Fundación Pública Galega de Medicina Xenómica, SERGAS, Grupo de Medicina Xenómica-USC, Health Research Institute of Santiago de Compostela (IDIS), Santiago de Compostela, Spain; 4Faculty of Health Sciences, Universidad Autónoma de Chile, Talca, Chile; 5Department of Respiratory Medicine, Kumamoto University Hospital, Kumamoto, Japan; 6Department of Oncology and Radiotherapy, Faculty of Medicine, Medical University of Gdańsk, Gdańsk, Poland Contributions: (I) Conception and design: Z Tu; (II) Administrative support: Z Tu; (III) Provision of study materials or patients: C Du, J Tang, Y Chen, J Zhao, D Zheng; (IV) Collection and assembly of data: C Du; (V) Data analysis and interpretation: C Du, J Cai; (VI) Manuscript writing: All authors; (VII) Final approval of manuscript: All authors. Correspondence to: Zhengliang Tu, PhD, MD. Department of Thoracic Surgery, The First Affiliated Hospital, Zhejiang University School of Medicine, 1367 West Wenyi Rd., Hangzhou 311121, China. Email: [tuzhl@sina.com](tuzhl@sina.com). Background: Aberrant methylation plays an essential role in early cancer development. In this study, we investigated methylation patterns in lung squamous cell carcinoma (LUSC) and matched non-tumor tissue and plasma samples to evaluate the potential of these patterns in the diagnosis ofLUSC. Methods: The study group included 49 patients with stage I–III LUSC. We collected resected tumor tissue, paired peritumoral tissue, distant normal tissue, and corresponding plasma samples. A bespoke lung cancer bisulfite sequencing panel was used to profile the methylation level. Another 48 healthy volunteers provided control plasma samples. Results: Peritumoral and distant normal tissues presented similar methylation signatures, distinct from those in tumor tissue samples. A comparison of methylation profiles led to the identification of 871 tumor-specific differentially methylated blocks, including 847 hypermethylated and 24 hypomethylated blocks (adjusted P value <0.05). All top-ranked blocks were tumor-related. Tissue samples were analyzed for field cancerizationto identify progressively aggravating aberrant methylations during tumor initiation and development. The analysis revealed that 221 blocks presented a stepwise increase in methylation levels, while seven blocks presented a stepwise decrease in methylation pattern as the sampling drew nearer to the tumor. The malignant contaminated ratio (MCR) confirmed the presence of distinct methylation patterns between tumor and peritumoral tissue samples. We then constructed a diagnostic panel using a combined diagnostic score of cellfree DNA (cfDNA) that showed high sensitivity and specificity. The healthy controls had a significantly lower combined diagnostic score (cd-score) than LUSC patients. Additionally, based on the methylation profiles, LUSC could be classified into two subgroups, C1 and C2 . The methylation profile of the C2 group was not distinct from the healthy controls, which had a significantly lower cd-score than did the C1 group. Conclusions: LUSC-specific methylation patterns could potentially discriminate between peritumoral tissue, distant normal tumor tissue, and tumor tissues. This preliminary study also supported the potential utility of cfDNA methylation analysis in diagnosing LUSC. Keywords: Methylation patterns; lung squamous cell carcinoma (LUSC); combined diagnostic score (cd-score); cell-free DNA (cfDNA) Submitted Dec 01, 2023. Accepted for publication Jan 17, 2

FAQs

What was the main goal of the study?

To evaluate whether methylation patterns in lung squamous cell carcinoma (LUSC) and matched non-tumor samples can support LUSC diagnosis, including the potential of cell-free DNA (cfDNA) methylation analysis.

How were methylation profiles generated and compared?

Tumor tissue, peritumoral tissue, distant normal tissue, and corresponding plasma samples were collected from LUSC patients and profiled using a bespoke lung cancer bisulfite sequencing panel; control plasma came from healthy volunteers.

What diagnostic value did the researchers observe?

A cfDNA-based diagnostic panel using a combined diagnostic score (cd-score) showed high sensitivity and specificity, and LUSC patients had significantly higher cd-scores than healthy controls.

Sep 21, 2026

Uploaded by Anda