Bayer HealthCare Pharmaceuticals, Whippany, New Jersey.
Bayer AG, Berlin, Germany.
Gastroenterology. 2019 May;156(6):1731-1741. doi: 10.1053/j.gastro.2019.01.261. Epub 2019 Feb 6.
BACKGROUND & AIMS: In a phase 3 trial (RESORCE), regorafenib increased overall survival compared with placebo in patients with hepatocellular carcinoma (HCC) previously treated with sorafenib. In an exploratory study, we analyzed plasma and tumor samples from study participants to identify genetic, microRNA (miRNA), and protein biomarkers associated with response to regorafenib.
We obtained archived tumor tissues and baseline plasma samples from patients with HCC given regorafenib in the RESORCE trial. Baseline plasma samples from 499 patients were analyzed for expression of 294 proteins (DiscoveryMAP) and plasma samples from 349 patients were analyzed for levels of 750 miRNAs (miRCURY miRNA PCR). Tumor tissues from 7 responders and 10 patients who did not respond (progressors) were analyzed by next-generation sequencing (FoundationOne). Forty-six tumor tissues were analyzed for expression patterns of 770 genes involved in oncogenic and inflammatory pathways (PanCancer Immune Profiling). Associations between plasma levels of proteins and miRNAs and response to treatment (overall survival and time to progression) were evaluated using a Cox proportional hazards model.
Decreased baseline plasma concentrations of 5 of 266 evaluable proteins (angiopoietin 1, cystatin B, the latency-associated peptide of transforming growth factor beta 1, oxidized low-density lipoprotein receptor 1, and C-C motif chemokine ligand 3; adjusted P ≤ .05) were significantly associated with increased overall survival time after regorafenib treatment. Levels of these 5 proteins, which have roles in inflammation and/or HCC pathogenesis, were not associated with survival independently of treatment. Only 20 of 499 patients had high levels and a reduced survival time. Plasma levels of α-fetoprotein and c-MET were associated with poor outcome (overall survival) independently of regorafenib treatment only. We identified 9 plasma miRNAs (MIR30A, MIR122, MIR125B, MIR200A, MIR374B, MIR15B, MIR107, MIR320, and MIR645) whose levels significantly associated with overall survival time with regorafenib (adjusted P ≤ .05). Functional analyses of these miRNAs indicated that their expression level associated with increased overall survival of patients with tumors of the Hoshida S3 subtype. Next-generation sequencing analyses of tumor tissues revealed 49 variants in 27 oncogenes or tumor suppressor genes. Mutations in CTNNB1 were detected in 3 of 10 progressors and VEGFA amplification in 1 of 7 responders.
We identified expression patterns of plasma proteins and miRNAs that associated with increased overall survival times of patients with HCC following treatment with regorafenib in the RESORCE trial. Levels of these circulating biomarkers and genetic features of tumors might be used to identify patients with HCC most likely to respond to regorafenib. ClinicalTrials.gov number NCT01774344. NCBI GEO accession numbers: mRNA data (NanoString): GSE119220; miRNA data (Exiqon): GSE119221.
在一项 3 期试验（RESORCE）中，regorafenib 相较于安慰剂增加了索拉非尼治疗后的肝癌（HCC）患者的总生存期。在一项探索性研究中，我们分析了研究参与者的血浆和肿瘤样本，以鉴定与regorafenib 反应相关的遗传、微小 RNA（miRNA）和蛋白质生物标志物。
我们从 RESORCE 试验中接受regorafenib 的 HCC 患者获得了存档的肿瘤组织和基线血浆样本。对 499 例患者的基线血浆样本进行了 294 种蛋白质的表达分析（DiscoveryMAP），对 349 例患者的血浆样本进行了 750 种 miRNA（miRCURY miRNA PCR）水平分析。对 7 名应答者和 10 名未应答（进展者）的患者的肿瘤组织进行了下一代测序（FoundationOne）分析。对 46 例肿瘤组织进行了涉及致癌和炎症途径的 770 种基因表达模式分析（PanCancer Immune Profiling）。使用 Cox 比例风险模型评估血浆蛋白和 miRNA 水平与治疗反应（总生存期和无进展生存期）之间的关联。
266 种可评估蛋白中有 5 种（血管生成素 1、半胱氨酸蛋白酶抑制剂 B、转化生长因子β 1 的潜伏期相关肽、氧化型低密度脂蛋白受体 1 和 C-C 基序趋化因子配体 3；调整后的 P ≤.05）的基线血浆浓度降低与 regorafenib 治疗后总生存期的延长显著相关。这些在炎症和/或 HCC 发病机制中起作用的 5 种蛋白质的水平与治疗无关的生存无关。只有 20 例患者（499 例患者中有 20 例）存在高水平和较短的生存时间。α-胎蛋白和 c-MET 的血浆水平与不良预后（总生存期）相关，仅与 regorafenib 治疗无关。我们鉴定了 9 种与总生存期相关的血浆 miRNA（MIR30A、MIR122、MIR125B、MIR200A、MIR374B、MIR15B、MIR107、MIR320 和 MIR645）（调整后的 P ≤.05）。这些 miRNA 的功能分析表明，其表达水平与 Hoshida S3 亚型肿瘤患者的总生存期延长相关。肿瘤组织的下一代测序分析显示，在 27 个癌基因或肿瘤抑制基因中发现了 49 个变异。在 10 名进展者中检测到 CTNNB1 突变，在 7 名应答者中检测到 VEGFA 扩增。
我们鉴定了 HCC 患者接受 RESORCE 试验中 regorafenib 治疗后与总生存期延长相关的血浆蛋白和 miRNA 表达模式。这些循环生物标志物的水平和肿瘤的遗传特征可能用于识别最有可能对 regorafenib 有反应的 HCC 患者。临床试验注册号：NCT01774344。NCBI GEO 注册号：mRNA 数据（NanoString）：GSE119220；miRNA 数据（Exiqon）：GSE119221。
