Poel Dennis, Rustenburg François, Sie Daoud, van Essen Hendrik F, Eijk Paul P, Bloemena Elisabeth, Elhorst Benites Teresita, van den Berg Madeleine C, Vergeer Marije R, Leemans René C, Buffart Tineke E, Ylstra Bauke, Brakenhoff Ruud H, Verheul Henk M, Voortman Jens
Amsterdam UMC, Vrije Universiteit Amsterdam, Department of Medical Oncology, Cancer Center Amsterdam, the Netherlands; Department of Medical Oncology, Radboud University Medical Center, Nijmegen, the Netherlands.
Amsterdam UMC, Vrije Universiteit Amsterdam, Department of Neurosurgery, Cancer Center Amsterdam, the Netherlands.
Oral Oncol. 2020 Jun 22;109:104851. doi: 10.1016/j.oraloncology.2020.104851.
The majority of patients with locally advanced larynx or hypopharynx squamous cell carcinoma are treated with organ-preserving chemoradiotherapy (CRT). Clinical outcome following CRT varies greatly. We hypothesized that tumor microRNA (miRNA) expression is predictive for outcome following CRT.
Next-generation sequencing (NGS) miRNA profiling was performed on 37 formalin-fixed paraffin-embedded (FFPE) tumor samples. Patients with a recurrence-free survival (RFS) of less than 2 years and patients with late/no recurrence within 2 years were compared by differential expression analysis. Tumor-specific miRNAs were selected based on normal mucosa miRNA expression data from The Cancer Genome Atlas database. A model was constructed to predict outcome using group-regularized penalized logistic ridge regression. Candidate miRNAs were validated by RT-qPCR in the initial sample set as well as in 46 additional samples.
Thirteen miRNAs were differentially expressed (p < 0.05, FDR < 0.1) according to outcome group. Initial class prediction in the NGS cohort (n = 37) resulted in a model combining five miRNAs and disease stage, able to predict CRT outcome with an area under the curve (AUC) of 0.82. In the RT-qPCR cohort (n = 83), 25 patients (30%) experienced early recurrence (median RFS 8 months; median follow-up 42 months). Class prediction resulted in a model combining let-7i-5p, miR-192-5p and disease stage, able to discriminate patients with good versus poor clinical outcome (AUC:0.80).
The combined miRNA expression and disease stage prediction model for CRT outcome is superior to using either factor alone. This study indicates NGS miRNA profiling using FFPE specimens is feasible, resulting in clinically relevant biomarkers.
大多数局部晚期喉或下咽鳞状细胞癌患者接受保留器官的放化疗（CRT）。CRT后的临床结果差异很大。我们假设肿瘤微小RNA（miRNA）表达可预测CRT后的结果。
对37份福尔马林固定石蜡包埋（FFPE）肿瘤样本进行下一代测序（NGS）miRNA分析。通过差异表达分析比较无复发生存期（RFS）小于2年的患者和2年内晚期/无复发的患者。根据来自癌症基因组图谱数据库的正常黏膜miRNA表达数据选择肿瘤特异性miRNA。使用组正则化惩罚逻辑岭回归构建预测结果的模型。候选miRNA在初始样本集以及另外46个样本中通过RT-qPCR进行验证。
根据结果分组，有13种miRNA差异表达（p<0.05，FDR<0.1）。NGS队列（n = 37）中的初始分类预测产生了一个结合5种miRNA和疾病分期的模型，能够以0.82的曲线下面积（AUC）预测CRT结果。在RT-qPCR队列（n = 83）中，25名患者（30%）出现早期复发（中位RFS 8个月；中位随访42个月）。分类预测产生了一个结合let-7i-5p、miR-192-5p和疾病分期的模型，能够区分临床结果良好与不良的患者（AUC：0.80）。
用于CRT结果的miRNA表达与疾病分期联合预测模型优于单独使用任何一个因素。本研究表明使用FFPE标本进行NGS miRNA分析是可行的，可产生具有临床相关性的生物标志物。
