A detection panel of novel methylated DNA markers for malignant pleural effusion

Chaonan Liang; Nan Liu; Qin Zhang; Mingming Deng; Jiangwei Ma; Jingwen Lu; Yan Yin; Jian Wang; Yuan Miao; Bin She; Qingchang Li; Gang Hou

doi:10.3389/fonc.2022.967079

A detection panel of novel methylated DNA markers for malignant pleural effusion

Front Oncol. 2022 Sep 13:12:967079. doi: 10.3389/fonc.2022.967079. eCollection 2022.

Authors

Chaonan Liang^{1

2}, Nan Liu³, Qin Zhang², Mingming Deng⁴, Jiangwei Ma², Jingwen Lu², Yan Yin², Jian Wang³, Yuan Miao³, Bin She⁵, Qingchang Li³, Gang Hou⁴

Affiliations

¹ Department of Cardio-Pulmonary Function, Henan Provincial People's Hospital, Zhengzhou University People's Hospital, Henan University People's Hospital, Zhengzhou, Henan, China.
² Department of Pulmonary and Critical Care Medicine, The First Hospital of China Medical University, Shenyang, China.
³ Department of Pathology, The First Hospital and College of Basic Medical Sciences, China Medical University, Shenyang, China.
⁴ Department of Pulmonary and Critical Care Medicine, China-Japan Friendship Hospital, National Center of Respiratory Medicine, Institute of Respiratory Medicine, Chinese Academy of Medical Sciences, National Clinical Research Center for Respiratory Diseases, Beijing, China.
⁵ Academic Development, Tellgen Corporation, Shanghai, China.

Abstract

Background: Cytology remains the gold standard for the detection of malignant cells in pleural effusion. However, its sensitivity is limited. The aim of this study was to establish a novel panel of cancer-specific methylated genes for the differential diagnosis of malignant pleural effusion (MPE).

Methods: A cohort of 100 cancer patients (68 lung cancer, 32 other malignant tumors) and 48 patients with benign disease presenting with pleural effusion was prospectively enrolled. Pleural effusion was evaluated by means of cytopathological investigation and DNA methylation of SHOX2, RASSF1A, SEPTIN9 and HOXA9 in the cellular fraction. DNA methylation in bisulfite-converted DNA was determined using quantitative methylation-specific real-time PCR (MS-PCR). Cytopathological and DNA methylation results were evaluated with regard to the final clinical diagnosis.

Results: The LungMe^® SHOX2 and RASSF1A Assay (Tellgen Corporation, China) has been reported to be highly sensitive and specific for lung cancer using bronchial aspirates. As expected, LungMe^® detected metastases of lung cancer (sensitivity: 76.5%) as well as metastases of other malignant tumors (sensitivity: 68.8%). OncoMe, a novel combination of SHOX2, RASSF1A, SEPTIN9 and HOXA9 methylation, led to an additional 11% increase in the detection rate of MPE, resulting in a sensitivity of 85% and a specificity of 96%. Overall, OncoMe showed a higher positive detection rate in SCLC (100%), LUAC (87%), OC (100%), BC (92.9%), GC (80.0%), and MESO (80%) than in LUSC (50%). Cytopathological analyses only detected 23 positive samples, which were all positively measured by both LungMe^® and OncoMe.

Conclusion: OncoMe has potential for use as a biomarker for the detection of MPE, even not limited to lung cancer.

Keywords: DNA methylation; HOXA9; RASSF1A; SEPTIN9; SHOX2; biomarkers; pleural effusion.