A blood-based prognostic liver secretome signature and long-term hepatocellular carcinoma risk in advanced liver fibrosis

Background Accurate non-invasive prediction of long-term hepatocellular carcinoma (HCC) risk in advanced liver fibrosis is urgently needed for cost-effective HCC screening; however, this currently remains an unmet need. Methods A serum-protein-based prognostic liver secretome signature (PLSec) was bioinformatically derived from previously validated hepatic transcriptome signatures and optimized in 79 patients with advanced liver fibrosis. We independently validated PLSec for HCC risk in 331 cirrhosis patients with mixed etiologies (validation set 1 [V1], median follow-up 4.5 years) and thereafter developed a composite risk score including clinical prognostic variables. The score was then validated in two independent cohorts: validation set 2 (V2): 164 patients with advanced liver fibrosis due to hepatitis C virus (HCV) infection cured after direct-acting antiviral therapy (nested case-control series, median follow-up 4.5 years); validation set 3 (V3): 146 patients with advanced liver fibrosis with successfully-treated HCC and cured HCV infection (median follow-up 2.9 years). Findings An 8-protein blood-based PLSec recapitulated transcriptome-based hepatic HCC risk status. In V1, PLSec was significantly associated with incident HCC risk (adjusted hazard ratio [aHR], 2.35; 95% confidence interval [CI], 1.30-4.23). A composite score with serum alpha-fetoprotein (PLSec-AFP) was defined in V1, and validated in V2 (adjusted odds ratio, 3.80 [95%CI, 1.66-8.66]) and V3 (aHR, 3.08 [95%CI, 1.78-5.31]; c-index, 0.74). PLSec-AFP outperformed AFP alone (Brier score, 0.165 vs. 0.186 in V2; 0.196 vs. 0.206 in V3, respectively). Conclusions The blood-based PLSec-AFP can accurately stratify patients with advanced liver fibrosis for long-term HCC risk and thereby guide risk-based tailored HCC screening.背景 实现晚期肝纤维化患者长期肝细胞癌（hepatocellular carcinoma, HCC）风险的精准无创预测，是开展成本效益比最优的HCC筛查的迫切需求，但目前这一需求仍未得到满足。 方法 本研究通过生物信息学分析，从已验证的肝脏转录组特征中衍生出基于血清蛋白的预后肝脏分泌组特征（prognostic liver secretome signature, PLSec），并在79例晚期肝纤维化患者中完成模型优化。我们在331例混合病因肝硬化患者中独立验证了PLSec的HCC风险预测效能（验证集1[V1]，中位随访时长4.5年）；随后构建了整合临床预后变量的综合风险评分。该综合评分随后在两个独立队列中完成验证：验证集2（V2）：164例因丙型肝炎病毒（hepatitis C virus, HCV）感染接受直接抗病毒治疗后获得治愈的晚期肝纤维化患者（嵌套病例对照系列，中位随访时长4.5年）；验证集3（V3）：146例合并经成功治疗的肝细胞癌且丙型肝炎病毒感染已治愈的晚期肝纤维化患者（中位随访时长2.9年）。 结果 本研究获得的8蛋白血液PLSec可复现基于转录组的肝脏HCC风险状态。在V1队列中，PLSec与新发HCC风险显著相关（校正风险比（adjusted hazard ratio, aHR）：2.35；95%置信区间（95% confidence interval, 95%CI）：1.30~4.23）。我们在V1队列中定义了结合血清甲胎蛋白（alpha-fetoprotein, AFP）的综合评分（PLSec-AFP），并在V2队列（校正比值比：3.80 [95%CI：1.66~8.66]）与V3队列（校正风险比（adjusted hazard ratio, aHR）：3.08 [95%CI：1.78~5.31]；一致性指数（concordance index, c-index）：0.74）中完成验证。PLSec-AFP的预测性能优于单纯AFP检测（V2队列中布里尔评分（Brier score）分别为0.165 vs 0.186；V3队列中分别为0.196 vs 0.206）。 结论 基于血液的PLSec-AFP可精准分层晚期肝纤维化患者的长期HCC风险，从而指导基于风险分层的个体化HCC筛查。