Department of General Surgery, Zhongshan Hospital, Fudan University, Shanghai, China.
Shanghai Engineering Research Center of Colorectal Cancer Minimally Invasive Technology, Shanghai, China.
Clin Cancer Res. 2024 Aug 1;30(15):3337-3348. doi: 10.1158/1078-0432.CCR-24-0199.
Detection of colorectal carcinomas at a time when there are more treatment options is associated with better outcomes. This prospective case-control study assessed the 5-hydroxymethylcytosine (5hmC) biomarkers in circulating cell-free DNA (cfDNA) for early detection of colorectal carcinoma and advanced adenomas (AA).
Plasma cfDNA samples from 2,576 study participants from the multicenter METHOD-2 study (NCT03676075) were collected, comprising patients with newly diagnosed colorectal carcinoma (n = 1,074), AA (n = 356), other solid tumors (n = 80), and non-colorectal carcinoma/AA controls (n = 1,066), followed by genome-wide 5hmC profiling using the 5hmC-Seal technique and the next-generation sequencing. A weighted diagnostic model for colorectal carcinoma (stage I-III) and AA was developed using the elastic net regularization in a discovery set and validated in independent samples.
Distribution of 5hmC in cfDNA reflected gene regulatory relevance and tissue of origin. Besides being confirmed in internal validation, a 96-gene model achieved an area under the curve (AUC) of 90.7% for distinguishing stage I-III colorectal carcinoma from controls in 321 samples from multiple centers for external validation, regardless of primary location or mutation status. This model also showed cancer-type specificity as well as high capacity for distinguishing AA from controls with an AUC of 78.6%. Functionally, differential 5hmC features associated with colorectal carcinoma and AA demonstrated relevance to colorectal carcinoma biology, including pathways such as calcium and MAPK signaling.
Genome-wide mapping of 5hmC in cfDNA shows promise as a highly sensitive and specific noninvasive blood test to be integrated into screening programs for improving early detection of colorectal carcinoma and high-risk AA.
在有更多治疗选择的情况下检测结直肠癌与更好的结果相关。这项前瞻性病例对照研究评估了循环无细胞游离 DNA (cfDNA) 中的 5-羟甲基胞嘧啶 (5hmC) 生物标志物，以早期检测结直肠癌和高级腺瘤 (AA)。
从多中心 METHOD-2 研究（NCT03676075）的 2576 名研究参与者中收集了血浆 cfDNA 样本，包括新诊断的结直肠癌患者（n=1074）、AA 患者（n=356）、其他实体瘤患者（n=80）和非结直肠癌/AA 对照组（n=1066），随后使用 5hmC-Seal 技术和下一代测序进行全基因组 5hmC 分析。使用弹性网络正则化在发现集中开发用于结直肠癌（I-III 期）和 AA 的加权诊断模型，并在独立样本中进行验证。
cfDNA 中 5hmC 的分布反映了基因调控的相关性和组织起源。除了在内部验证中得到证实外，一个包含 96 个基因的模型在来自多个中心的 321 个样本中用于外部验证时，对于区分 I-III 期结直肠癌与对照的曲线下面积（AUC）达到 90.7%，无论原发部位或突变状态如何。该模型还表现出癌症类型特异性，以及区分 AA 与对照的高能力，AUC 为 78.6%。功能上，与结直肠癌和 AA 相关的差异 5hmC 特征与结直肠癌生物学相关，包括钙和 MAPK 信号等途径。
cfDNA 中 5hmC 的全基因组图谱显示出作为一种高度敏感和特异的非侵入性血液检测方法的潜力，可整合到筛查计划中，以提高结直肠癌和高危 AA 的早期检测。
