California NanoSystems Institute, Crump Institute for Molecular Imaging, Department of Molecular and Medical Pharmacology , University of California, Los Angeles , Los Angeles , California , USA.
Key Laboratory for Nano-Bio Interface, Suzhou Institute of Nano-Tech and Nano-Bionics , University of Chinese Academy of Sciences, Chinese Academy of Sciences , Suzhou , People's Republic of China.
Hepatology. 2023 Mar 1;77(3):774-788. doi: 10.1002/hep.32692. Epub 2023 Feb 17.
The sensitivity of current surveillance methods for detecting early-stage hepatocellular carcinoma (HCC) is suboptimal. Extracellular vesicles (EVs) are promising circulating biomarkers for early cancer detection. In this study, we aim to develop an HCC EV-based surface protein assay for early detection of HCC.
Tissue microarray was used to evaluate four potential HCC-associated protein markers. An HCC EV surface protein assay, composed of covalent chemistry-mediated HCC EV purification and real-time immuno-polymerase chain reaction readouts, was developed and optimized for quantifying subpopulations of EVs. An HCC EV ECG score, calculated from the readouts of three HCC EV subpopulations ( E pCAM + CD63 + , C D147 + CD63 + , and G PC3 + CD63 + HCC EVs), was established for detecting early-stage HCC. A phase 2 biomarker study was conducted to evaluate the performance of ECG score in a training cohort ( n = 106) and an independent validation cohort ( n = 72).Overall, 99.7% of tissue microarray stained positive for at least one of the four HCC-associated protein markers (EpCAM, CD147, GPC3, and ASGPR1) that were subsequently validated in HCC EVs. In the training cohort, HCC EV ECG score demonstrated an area under the receiver operating curve (AUROC) of 0.95 (95% confidence interval [CI], 0.90-0.99) for distinguishing early-stage HCC from cirrhosis with a sensitivity of 91% and a specificity of 90%. The AUROCs of the HCC EV ECG score remained excellent in the validation cohort (0.93; 95% CI, 0.87-0.99) and in the subgroups by etiology (viral: 0.95; 95% CI, 0.90-1.00; nonviral: 0.94; 95% CI, 0.88-0.99).
HCC EV ECG score demonstrated great potential for detecting early-stage HCC. It could augment current surveillance methods and improve patients' outcomes.
目前用于检测早期肝细胞癌（HCC）的监测方法的灵敏度并不理想。细胞外囊泡（EVs）是癌症早期检测有前途的循环生物标志物。在这项研究中，我们旨在开发一种基于 HCC EV 的表面蛋白检测方法，用于早期检测 HCC。
使用组织微阵列评估了四个潜在的 HCC 相关蛋白标志物。开发了一种 HCC EV 表面蛋白检测方法，由共价化学介导的 HCC EV 纯化和实时免疫聚合酶链反应读数组成，用于定量 EV 亚群。根据三个 HCC EV 亚群（ E pCAM + CD63 + ， C D147 + CD63 + ，和 G PC3 + CD63 + HCC EVs）的读数计算 HCC EV ECG 评分，用于检测早期 HCC。进行了一项 2 期生物标志物研究，以评估 ECG 评分在训练队列（ n = 106）和独立验证队列（ n = 72）中的性能。总体而言，组织微阵列上至少有四种 HCC 相关蛋白标志物（EpCAM、CD147、GPC3 和 ASGPR1）中的一种染色阳性，这些标志物随后在 HCC EVs 中得到验证。在训练队列中，HCC EV ECG 评分在区分早期 HCC 与肝硬化方面的曲线下面积（AUROC）为 0.95（95%置信区间[CI]，0.90-0.99），灵敏度为 91%，特异性为 90%。在验证队列中，HCC EV ECG 评分的 AUROC 仍然很高（0.93；95%CI，0.87-0.99），在病因亚组中也是如此（病毒：0.95；95%CI，0.90-1.00；非病毒：0.94；95%CI，0.88-0.99）。
HCC EV ECG 评分在检测早期 HCC 方面具有很大的潜力。它可以增强当前的监测方法并改善患者的预后。
