通过结合血浆游离 DNA 中 5-羟甲基胞嘧啶特征和蛋白质生物标志物进行液体活检，用于诊断和预测肝细胞癌。

Liquid biopsy by combining 5-hydroxymethylcytosine signatures of plasma cell-free DNA and protein biomarkers for diagnosis and prognosis of hepatocellular carcinoma.

机构信息

The United Innovation of Mengchao Hepatobiliary Technology Key Laboratory of Fujian Province, Mengchao Hepatobiliary Hospital of Fujian Medical University, Fuzhou, P. R. China; Mengchao Med-X Center, Fuzhou University, Fuzhou, P. R. China.

Frontier Science Center for Disease Molecular Network, State Key Laboratory of Biotherapy, West China Hospital of Sichuan University, Chengdu, P. R. China.

出版信息

ESMO Open. 2021 Feb;6(1):100021. doi: 10.1016/j.esmoop.2020.100021. Epub 2021 Jan 25.

DOI:10.1016/j.esmoop.2020.100021

PMID:33508734

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7841321/

Abstract

BACKGROUND

Liquid biopsy based on 5-hydroxymethylcytosine (5hmC) signatures of plasma cell-free DNA (cfDNA) originating from tumor cells provides a novel approach for early diagnosis in hepatocellular carcinoma (HCC). Here, we sought to develop a reliable model using cfDNA 5hmC signatures and protein biomarkers for diagnosis and prognosis of HCC.

PATIENTS AND METHODS

We carried out genome-wide 5hmC sequencing of cfDNA samples collected from 165 healthy volunteers, 62 liver cirrhosis (LC) patients and 135 HCC patients. A sensitive 5hmC diagnostic model was developed based on 5hmC signatures selected by sparse Partial Least Squares Discriminant Analysis and cross-validation to define the weighted diagnostic score (wd-score). Then, we combined protein biomarkers with the wd-score to build a more robust score (HCC score) by logistic regression.

RESULTS

The distribution pattern of differential 5hmC regions could clearly distinguish HCC patients, LC patients and healthy volunteers. The wd-score based on 64 5hmC signatures in cfDNA achieves 93.24% of area under the curve (AUC) to distinguish HCC patients from non-HCC patients, and the HCC score by combing protein biomarkers achieves 92.75% of AUC to distinguish HCC patients from LC patients. Meanwhile, the HCC score showed high capacity for screening high recurrence risk patients after receiving surgical resection, and appeared to be an independent indicator for both relapse-free survival (P = 0.00865) and overall survival (P = 0.000739). Furthermore, the values of the HCC score in patients' longitudinal plasma samples were positively associated with tumor burden dynamics during follow-up.

CONCLUSION

We have developed and validated a novel non-invasive liquid biopsy strategy for HCC diagnosis, prognosis and surveillance during HCC progression.

摘要

背景

基于源自肿瘤细胞的血浆无细胞 DNA（cfDNA）中 5-羟甲基胞嘧啶（5hmC）特征的液体活检为肝细胞癌（HCC）的早期诊断提供了一种新方法。在这里，我们试图使用 cfDNA 5hmC 特征和蛋白质生物标志物为 HCC 的诊断和预后开发一种可靠的模型。

患者和方法

我们对 165 名健康志愿者、62 名肝硬化（LC）患者和 135 名 HCC 患者的 cfDNA 样本进行了全基因组 5hmC 测序。通过稀疏偏最小二乘判别分析和交叉验证选择的 5hmC 特征来开发敏感的 5hmC 诊断模型，以定义加权诊断评分（wd-score）。然后，我们通过逻辑回归将蛋白质生物标志物与 wd-score 结合起来，构建更稳健的评分（HCC 评分）。

结果

差异 5hmC 区域的分布模式可以清楚地区分 HCC 患者、LC 患者和健康志愿者。基于 cfDNA 中 64 个 5hmC 特征的 wd-score 可区分 HCC 患者和非 HCC 患者，曲线下面积（AUC）为 93.24%，结合蛋白质生物标志物的 HCC 评分可区分 HCC 患者和 LC 患者，AUC 为 92.75%。同时，该 HCC 评分在接受手术切除后对筛选高复发风险患者具有较高的能力，并且似乎是无复发生存（P=0.00865）和总生存（P=0.000739）的独立指标。此外，患者纵向血浆样本中 HCC 评分的值与随访期间肿瘤负担动态呈正相关。