The ability to detect several types of cancer using a non-invasive, blood-based test holds the potential to revolutionize oncology screening. We mined tumor methylation array data from the Cancer Genome Atlas (TCGA) covering 14 cancer types and identified two novel, broadly-occurring methylation markers atTLX1andGALR1. To evaluate their performance as a generalized blood-based screening approach, along with our previously reported methylation biomarker,ZNF154, we rigorously assessed each marker individually or combined. Utilizing TCGA methylation data and applying logistic regression models within each individual cancer type, we found that the three-marker combination significantly increased the average area under the ROC curve (AUC) across the 14 tumor types compared to single markers (p= 1.158 × 10−10; Friedman test). Furthermore, we simulated dilutions of tumor DNA into healthy blood cell DNA and demonstrated increased AUC of combined markers across all dilution levels. Finally, we evaluated assay performance in bisulfite sequenced DNA from patient tumors and plasma, including early-stage samples. When combining all three markers, the assay correctly identified nine out of nine lung cancer plasma samples. In patient plasma from hepatocellular carcinoma,ZNF154alone yielded the highest combined sensitivity and specificity values averaging 68% and 72%, whereas multiple markers could achieve higher sensitivity or specificity, but not both. Altogether, this study presents a comprehensive pipeline for the identification, testing, and validation of multi-cancer methylation biomarkers with a considerable potential for detecting a broad range of cancer types in patient blood samples.
利用非侵入性血液检测技术检测多种癌症的能力，有望彻底改变肿瘤筛查模式。本研究通过挖掘癌症基因组图谱（TCGA）中涵盖14种癌症类型的肿瘤甲基化阵列数据，在TLX1和GALR1基因位点鉴定出两个新型的广谱甲基化标志物。为评估其作为通用血液筛查方法的性能，我们结合先前报道的甲基化标志物ZNF154，对单个标志物及组合标志物进行了系统评估。利用TCGA甲基化数据并在各癌种内建立逻辑回归模型，我们发现三标志物组合相较于单标志物，在14种肿瘤类型中显著提高了受试者工作特征曲线下面积（AUC）的平均值（p=1.158×10⁻¹⁰；Friedman检验）。此外，通过模拟肿瘤DNA在健康血细胞DNA中的稀释梯度，我们证实组合标志物在所有稀释水平下均能提升AUC值。最后，我们在患者肿瘤组织与血浆（含早期样本）的亚硫酸氢盐测序DNA中验证了检测性能：三标志物组合成功检出全部9例肺癌血浆样本；在肝细胞癌患者血浆中，单独使用ZNF154可获得平均68%的敏感性与72%的特异性，而多标志物组合虽能提升单项指标，但未能同时优化两者。本研究构建了一套完整的多癌种甲基化标志物鉴定、测试与验证流程，在利用患者血液样本检测广谱癌症类型方面展现出显著潜力。
Evaluating Stacked Methylation Markers for Blood-Based Multicancer Detection
肿瘤(癌症)患者之家