BACKGROUND & AIMS Cirrhosis and chronic inflammation precede development of hepatocellular carcinoma (HCC) in approximately 80% of cases. We investigated immune-related gene expression patterns in liver tissues surrounding early-stage HCCs and chemopreventive agents that might alter these patterns to prevent liver tumorigenesis. METHODS We analyzed gene expression profiles of nontumor liver tissues from 392 patients with early-stage HCC (training set, N = 167 and validation set, N = 225) and liver tissue from patients with cirrhosis without HCC (N = 216, controls) to identify changes in expression of genes that regulate the immune response that could contribute to hepatocarcinogenesis. We defined 172 genes as markers for this deregulated immune response, which we called the immune-mediated cancer field (ICF). We analyzed the expression data of liver tissues from 216 patients with cirrhosis without HCC and investigated the association between this gene expression signature and development of HCC and outcomes of patients (median follow-up, 10 years). Human liver tissues were also analyzed by histology. C57BL/6J mice were given a single injection of diethylnitrosamine (DEN) followed by weekly doses of carbon tetrachloride to induce liver fibrosis and tumorigenesis. Mice were then orally given the multiple tyrosine inhibitor nintedanib or vehicle (controls); liver tissues were collected and histology, transcriptome, and protein analyses were performed. We also analyzed transcriptomes of liver tissues collected from mice on a choline-deficient high-fat diet, which developed chronic liver inflammation and tumors, orally given aspirin and clopidogrel or the anti-inflammatory agent sulindac vs mice on a chow (control) diet. RESULTS We found the ICF gene expression pattern in 50% of liver tissues from patients with cirrhosis without HCC and in 60% of nontumor liver tissues from patients with early-stage HCC. The liver tissues with the ICF gene expression pattern had 3 different features: increased numbers of effector T cells; increased expression of genes that suppress the immune response and activation of transforming growth factor β signaling; or expression of genes that promote inflammation and activation of interferon gamma signaling. Patients with cirrhosis and liver tissues with the immunosuppressive profile (10% of cases) had a higher risk of HCC (hazard ratio, 2.41; 95% confidence interval, 1.21-4.80). Mice with chemically induced fibrosis or diet-induced steatohepatitis given nintedanib or aspirin and clopidogrel down-regulated the ICF gene expression pattern in liver and developed fewer and smaller tumors than mice given vehicle. CONCLUSIONS We identified an immune-related gene expression pattern in liver tissues of patients with early-stage HCC, called the ICF, that is associated with risk of HCC development in patients with cirrhosis. Administration of nintedanib or aspirin and clopidogrel to mice with chronic liver inflammation caused loss of this gene expression pattern and development of fewer and smaller liver tumors. Agents that alter immune regulatory gene expression patterns associated with carcinogenesis might be tested as chemopreventive agents in patients with cirrhosis.

中文翻译：

---

与人类肝细胞癌发展相关的免疫基因表达特征识别出对化学预防剂有反应的小鼠。

背景和目的 大约 80% 的病例中，肝硬化和慢性炎症先于肝细胞癌 (HCC) 的发展。我们研究了早期肝癌周围肝组织中免疫相关基因的表达模式，以及可能改变这些模式以预防肝脏肿瘤发生的化学预防剂。方法 我们分析了 392 名早期 HCC 患者的非肿瘤肝组织（训练集，N = 167 和验证集，N = 225）和肝硬化非 HCC 患者的肝组织（N = 216，对照）的基因表达谱，以确定调节可能导致肝癌发生的免疫反应的基因表达的变化。我们定义了 172 个基因作为这种失调的免疫反应的标记，我们将其称为免疫介导的癌症领域 (ICF)。我们分析了 216 名无 HCC 的肝硬化患者的肝组织表达数据，并研究了该基因表达特征与 HCC 发展和患者结局之间的关联（中位随访时间，10 年）。还通过组织学分析了人类肝脏组织。C57BL/6J 小鼠单次注射二乙基亚硝胺 (DEN)，然后每周注射一次四氯化碳，以诱导肝纤维化和肿瘤发生。然后给小鼠口服多重酪氨酸抑制剂尼达尼布或载体（对照组）；收集肝组织并进行组织学、转录组和蛋白质分析。我们还分析了从缺乏胆碱的高脂肪饮食的小鼠中收集的肝组织的转录组，这些小鼠出现了慢性肝脏炎症和肿瘤，口服阿司匹林和氯吡格雷或抗炎药舒林酸与饮食（对照）饮食的小鼠相比。结果我们发现 ICF 基因在 50% 的无 HCC 肝硬化患者的肝组织和 60% 的早期 HCC 患者的非肿瘤肝组织中存在表达模式。具有ICF基因表达模式的肝组织具有3个不同的特征：效应T细胞数量增加；抑制免疫反应和激活转化生长因子 β 信号传导的基因表达增加；或促进炎症和干扰素γ信号传导激活的基因的表达。肝硬化和肝组织具有免疫抑制特征的患者（10% 的病例）患 HCC 的风险较高（风险比，2.41；95% 置信区间，1.21-4.80）。患有化学诱导纤维化或饮食诱导脂肪性肝炎的小鼠，给予尼达尼布或阿司匹林和氯吡格雷后，下调了肝脏中 ICF 基因的表达模式，并且比给予媒介物的小鼠产生了更少和更小的肿瘤。结论 我们在早期 HCC 患者的肝组织中发现了一种免疫相关基因表达模式，称为 ICF，它与肝硬化患者发生 HCC 的风险相关。对患有慢性肝脏炎症的小鼠施用尼达尼布或阿司匹林和氯吡格雷会导致该基因表达模式丧失，并产生更少和更小的肝脏肿瘤。改变与癌发生相关的免疫调节基因表达模式的药物可以作为肝硬化患者的化学预防药物进行测试。