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Background  Lung (LC), prostate (PCa) and colorectal (CRC) cancers are the most incident in males worldwide. Despite recent advances, optimal population-based cancer screening methods remain an unmet need. Due to its early onset, cancer specificity and accessibility in body fluids, aberrant DNA promoter methylation might be a valuable minimally invasive tool for early cancer detection. Herein, we aimed to develop a minimally invasive methylation-based test for simultaneous early detection of LC, PCa and CRC in males, using liquid biopsies.    Results  Circulating cell-free DNA was extracted from 102 LC, 121 PCa and 100 CRC patients and 136 asymptomatic donors’ plasma samples. Sodium-bisulfite modification and whole-genome amplification was performed. Promoter methylation levels of  APC   me   , FOXA1   me   , GSTP1   me   , HOXD3   me  ,  RARβ2   me   , RASSF1A   me   , SEPT9   me   and  SOX17   me   were assessed by multiplex quantitative methylation-specific PCR.   SEPT9   me   and  SOX17   me   were the only biomarkers shared by all three cancer types, although they detected CRC with limited sensitivity. A “PanCancer” panel ( FOXA1   me   , RARβ2   me   and  RASSF1A   me  ) detected LC and PCa with 64% sensitivity and 70% specificity, complemented with “CancerType” panel ( GSTP1   me   and  SOX17   me  ) which discriminated between LC and PCa with 93% specificity, but with modest sensitivity. Moreover, a  HOXD3   me   and  RASSF1A   me   panel discriminated small cell lung carcinoma from non-small cell lung carcinoma with 75% sensitivity, 88% specificity, 6.5 LR+ and 0.28 LR–. An  APC   me   and  RASSF1A   me   panel independently predicted disease-specific mortality in LC patients.    Conclusions  We concluded that a DNA methylation-based test in liquid biopsies might enable minimally invasive screening of LC and PCa, improving patient compliance and reducing healthcare costs. Moreover, it might assist in LC subtyping and prognostication.

Early detection of the major male cancer types in blood-based liquid biopsies using a DNA methylation panel