淋巴结（LN）转移是确定口咽鳞状细胞癌（OPSCC）治疗和预后的重要因素。在这里，我们使用多区域测序比较了人类乳头瘤病毒（HPV）阳性OPSCC和HPV阴性OPSCC的原发灶和转移性LN的体细胞突变谱和克隆演化。我们对18名OPSCC患者（10名HPV阳性和8名HPV阴性），包括18个原发肿瘤样本、40个转移LN样本和18个正常组织样本进行了高深度全外显子测序（200x，76个样本）。在40个转移LN中，22个显示出淋巴结外侵（ENE）。HPV阳性OPSCC和HPV阴性OPSCC的突变谱与以前报道的相似。在HPV阴性OPSCC中，CDKN2A和TP53的体细胞突变经常被检测到。在HPV阳性OPSCC样本中，体细胞突变显示出APOBEC相关标志。转移LN的体细胞突变显示出与原发肿瘤不同的模式。转移过程中在WNT途径中获得的体细胞突变与ENE呈显著关系。原发灶和转移性LN的克隆演化分析表明，在某些情况下，每个转移LN来源于不同的原发肿瘤亚克隆。本文受版权保护。版权所有。

Lymph node (LN) metastasis is an important factor in determining the treatment and prognosis of oropharyngeal squamous cell carcinoma (OPSCC). Here, we compared the somatic mutational profiles and clonal evolution of primary and metastatic LNs using multiregion sequencing of human papilloma virus (HPV)-positive OPSCC and HPV-negative OPSCC. We performed high-depth whole-exome sequencing (200x) of 76 samples from 18 patients with OPSCC (10 HPV-positive and 8 HPV-negative), including 18 primary tumor samples, 40 metastatic LN samples and 18 normal tissue samples. Among 40 metastatic LNs, 22 showed extranodal extension (ENE). Mutation profiles of HPV-positive OPSCC and HPV-negative OPSCC were similar to those reported previously. Somatic mutations in CDKN2A and TP53 were frequently detected in HPV-negative OPSCC. Somatic mutations in HPV-positive OPSCC samples showed APOBEC-related signatures. Somatic mutations from metastatic LNs showed a different pattern than the primary tumor. Somatic mutations acquired in the WNT pathway during metastasis showed a significant relationship with ENE. Clonal evolution analysis of primary and metastatic LNs showed that, in some cases, each metastatic LN originated from a different primary tumor subclone.This article is protected by copyright. All rights reserved.