早期发现已被证明是降低结直肠癌（CRC）发病率和死亡率的最有效策略。然而，目前大多数筛查项目的参与率较低。血液检测可以提高筛查的依从性和结肠镜检查的选择。在这项研究中，我们在 433 个血清样本的多中心队列中进行了基于血清的 cfDNA 甲基化生物标志物的发现和验证，该生物标志物用于 CRC 筛查，其中包括健康对照、良性病变、晚期腺瘤 (AA) 和 CRC。首先，我们进行了表观基因组分析使用样本池方法使用 MmethylationEPIC 阵列进行全甲基化分析，然后对候选生物标志物进行强有力的优先排序，以检测晚期肿瘤（AN：AA 和 CRC）。然后，通过焦​​磷酸测序在独立的个体 cfDNA 样本中验证候选生物标志物。我们报告 GALNT9、UPF3A、WARS 和 LDB2 作为用于早期检测 AN 的新非侵入性生物标志物。 GALNT9/UPF3A 通过逻辑回归组合区分 AN 的敏感性为 78.8%，特异性为 100%，优于常用的粪便免疫化学测试和甲基化 SEPT9 血液测试。总体而言，本研究强调了 cfDNA 甲基化在 CRC 筛查中的实用性。我们的结果表明，甲基化 GALNT9/UPF3A 组合有潜力作为一种高度特异且灵敏的血液测试，用于筛查和早期检测 CRC。© 2023。BioMed Central Ltd.，Springer Nature 旗下公司。

Early detection has proven to be the most effective strategy to reduce the incidence and mortality of colorectal cancer (CRC). Nevertheless, most current screening programs suffer from low participation rates. A blood test may improve both the adherence to screening and the selection to colonoscopy. In this study, we conducted a serum-based discovery and validation of cfDNA methylation biomarkers for CRC screening in a multicenter cohort of 433 serum samples including healthy controls, benign pathologies, advanced adenomas (AA), and CRC.First, we performed an epigenome-wide methylation analysis with the MethylationEPIC array using a sample pooling approach, followed by a robust prioritization of candidate biomarkers for the detection of advanced neoplasia (AN: AA and CRC). Then, candidate biomarkers were validated by pyrosequencing in independent individual cfDNA samples. We report GALNT9, UPF3A, WARS, and LDB2 as new noninvasive biomarkers for the early detection of AN. The combination of GALNT9/UPF3A by logistic regression discriminated AN with 78.8% sensitivity and 100% specificity, outperforming the commonly used fecal immunochemical test and the methylated SEPT9 blood test.Overall, this study highlights the utility of cfDNA methylation for CRC screening. Our results suggest that the combination methylated GALNT9/UPF3A has the potential to serve as a highly specific and sensitive blood-based test for screening and early detection of CRC.© 2023. BioMed Central Ltd., part of Springer Nature.