Early Detection of Lung Cancer Using DNA Promoter Hypermethylation in Plasma and Sputum

Abstract

CT screening can reduce death from lung cancer. We sought to improve the diagnostic accuracy of lung cancer screening using ultrasensitive methods and a lung cancer-specific gene panel to detect DNA methylation in sputum and plasma. This is a case-control study of subjects with suspicious nodules on CT imaging. Plasma and sputum were obtained preoperatively. Cases ( = 150) had pathologic confirmation of node-negative (stages I and IIA) non-small cell lung cancer. Controls ( = 60) had non-cancer diagnoses. We detected promoter methylation using quantitative methylation-specific real-time PCR and methylation-on-beads for cancer-specific genes (, and ). DNA methylation was detected in plasma and sputum more frequently in people with cancer compared with controls ( < 0.001) for five of six genes. The sensitivity and specificity for lung cancer diagnosis using the best individual genes was 63% to 86% and 75% to 92% in sputum, respectively, and 65% to 76% and 74% to 84% in plasma, respectively. A three-gene combination of the best individual genes has sensitivity and specificity of 98% and 71% using sputum and 93% and 62% using plasma. Area under the receiver operating curve for this panel was 0.89 [95% confidence interval (CI), 0.80-0.98] in sputum and 0.77 (95% CI, 0.68-0.86) in plasma. Independent blinded random forest prediction models combining gene methylation with clinical information correctly predicted lung cancer in 91% of subjects using sputum detection and 85% of subjects using plasma detection. High diagnostic accuracy for early-stage lung cancer can be obtained using methylated promoter detection in sputum or plasma. .

Epigenetic biomarkers of promoter DNA methylation in the new era of cancer treatment

“…Digital PCR for CDO1 methylation (which can detect a 1/100 000 dilution of fully methylated CDO1 ) might be sufficient to detect cancer cells in plasma, as shown for VIMENTIN ( VIM ) gene hypermethylation in patients with CRC . However, CDO1 gene methylation was reported to be readily detected in the plasma of patients with lung cancer, where Q‐MSP detection of CDO1 methylation in plasma or serum showed 65% sensitivity with 74% specificity, and, when assessed in combination with the methylation of other HRMG, showed high sensitivity even in stage I disease. This extraordinarily high sensitivity of detection of cancer cells in plasma has been for the first time experienced in cancer clinics.…”

Aberrant methylation of mutL homolog 1 is associated with increased risk of non‐small cell lung cancer

“…A number of studies found a handful of genes with aberrant DNA methylation in lung cancers, implying a potential role of DNA methylation in the prediction for lung cancer. MLH1 is a tumor suppressor gene involved in DNA mismatch repair, which could correct the DNA replication .…”

Lung cancer early detection and health disparities: the intersection of epigenetics and ethnicity

“…The technique, known as methylation on beads (MOB), utilizes magnetic nanoparticles in an effort to combine DNA isolation and bisulfite treatment procedures into a single tube thereby increasing sample yield nearly 6-fold relative to the next most efficient kit-based assay (22). Since then, many markers have been identified and validated in bodily fluids with potential for use as independent or collaborative diagnostic tests in tandem with current screening methodologies (23)(24)(25)(26)(27)(28)(29).…”