预后不良的肾细胞癌伴淋巴结转移的分子特征

Molecular characteristics for poor prognosis related renal cell carcinoma with lymph metastases

摘要

目的：通过下一代测序（next-generation sequencing，NGS）技术分析肾细胞癌（简称肾癌）淋巴结转移的基因突变谱，发现与预后不良相关的分子特征，为肾癌机制的研究以及治疗提供新的思路。方法：对 31 例淋巴结转移的肾癌和 21 例非转移性肾癌患者的临床资料进行回顾性分析，同时从病理科调取肿瘤原发灶、淋巴结转移灶、远处转移灶的石蜡包埋组织共81例，使用 NGS 技术检测患者的基因突变谱，术后随访分析淋巴结转移与患者预后的关系。结果：与非转移性肾癌相比，淋巴结转移肾癌在肿瘤大小（P=0.006）、世界卫生组织（World Health Orga-nization，WHO）/国际泌尿病理协会（International Society of Urological Pathology，ISUP）分级（P=0.002）、T 分期（P= 0.003）、合并癌栓（P=0.025）上差异有统计学意义。最常见的突变基因是肿瘤抑制基因 VHL（38%）、 PBRM1 （22%）和 IGF2R（20%），且拷贝数变异与肿瘤转移相关，部分突变特征与已知的突变模式高度相似。淋巴结转移组和非转移组患者的突变频率存在差异，转移组大多数基因的突变频率较高，然而 Reactome 通路富集分析未在两组共有的富集途径上显示出差异有统计学意义。同一患者的肿瘤原发灶和转移灶之间有很强的一致性，基因组学指标[如纯度、倍性、加权基因组完整性指数（weighted-genomic integrity index，WGII）和肿瘤内异质性（intra-tumor heterogeneity，ITH）]及克隆、亚克隆组成分析进一步支持了这种一致性。无转移组的总生存率较高（P=0.041），且特定基因突变（如 IGF2R、JUN、EPHA5 和 FH）与较差的预后相关。淋巴结可能起到＂转移池＂的作用，促进肾癌的远处转移。结论：多基因 NGS 同时评估了多个相关标记物，揭示了淋巴结转移肾癌患者的基因改变谱，基于 NGS的分子分析可以帮助临床医生评估患者的预后，并发现新的机制及治疗靶点。

Abstract

Objective：Next-generation sequencing（NGS）technology was used to analyze the gene mutation profile of lymph node metastases in renal cell carcinoma，and the molecular characteristics asso-ciated with poor prognosis were found，providing new ideas for mechanism research and treatment. Methods：Retrospective clinical data collection was conducted on 31 patients with lymphoid metastatic renal cell carcinoma and 21 patients with non-metastatic renal cell carcinoma.A total of 81 formalin-fixed paraffin-embedded tissue samples were retrieved from the Department of Pathology，including primary tumor，lymph node metastasis，and distant metastasis samples.The gene mutation profiles of the patients were examined using next-generation sequencing technology.The patients were followed up to analyze the correlation between lymph node metastasis and patient prognosis. Results：The lymph node metastasis group showed differences in tumor size（P=0.006），World Health Organization（WHO）/International Society of Urological Pathology（ISUP）grade (P=0.002) ，T stage (P=0.003) and tumor thrombus (P=0.025) compared with non-metastatic renal cell carcinoma.The most commonly mutated genes in our cohort were the tumor suppressor genes VHL（38％），PBRM1（22％），and SETD2（20％）.More- over，copy number variations were associated with tumor metastasis，and some mutation features were highly similar to known mutation patterns.There was a difference in mutation frequency between the pa- tients in the metastasis group and samples in the non-metastasis group.The mutation frequency of most genes in the metastasis group was higher，however，Reactome pathway enrichment analysis did not show statistically significant differences in the shared enriched pathways between the two groups.There was a strong degree of concordance between the tumor＇s primary and metastatic foci in the same patient，and genomic indicators[such as purity，ploidy，weighted-genomic integrity index（WGII），and intra-tumor heterogeneity（ITH）]as well as clonal and subclonal composition analysis further supported this consis- tency.The overall survival（OS）was higher in the patients without metastases (P=0.041) ，and speci- fic gene mutations（such as IGF2R，JUN，EPHA5，and FH）were associated with poorer prognosis.To facilitate distant metastasis，lymph nodes might function as a＂metastatic pool＂. Conclusion：The multi- gene NGS evaluates multiple relevant markers simultaneously，revealing several genetic alterations in the patients with lymphatic metastatic renal cell carcinorma.NGS-based molecular analysis can assist clini- cians in assessing a patient＇s prognosis and identifying novel，potentially therapeutic mechanisms.

关键词

肾细胞癌 / 淋巴转移 / 高通量核苷酸序列分析 / DNA 突变分析 / 预后

Key words

Renal cell carcinoma / Lymphatic metastasis / High-throughput nucleotide Sequencing / DNA mutational analysis / Prognosis

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基金资助

国家自然科学基金(82273389)

北京市自然科学基金(7232212)