Janssen J, Andrade Barbosa B, Machado J C, Hofman P, Kim Y, Ylstra B, Radonic T
Department of Pathology, Cancer Center Amsterdam, Amsterdam UMC, Vrije Universiteit Amsterdam, Amsterdam, The Netherlands.
Institute of Molecular Pathology and Immunology of the University of Porto (IPATIMUP), Porto, France; Institute for Research and Innovation in Health (i3S), Porto, Portugal; Faculty of Medicine of the University of Porto, Porto, Portugal.
ESMO Open. 2025 May;10(5):105072. doi: 10.1016/j.esmoop.2025.105072. Epub 2025 May 14.
Next-generation sequencing (NGS) mutation panels are widely implemented and commonly applied to aid clonality classification for non-small-cell lung carcinoma (NSCLC) patients with multiple tumors. Performance of different NGS panels for clonality classification, however, remains unresolved.
We assembled 210 primary and metastatic pairs from lung adenocarcinoma (LUAD) and lung squamous-cell carcinomas (LUSC) of the TRACERx421 cohort for which gold standard clonality was confirmed by whole exome sequencing. We used four NGS panels ranging from 12 to 523 genes to determine clonality using the 2024 International Association for the Study of Lung Cancer (IASLC) recommendations.
With an oncogene panel, 30% LUAD and 74% LUSC pairs remained inconclusive with, respectively, 2% and 0% misclassified. Addition of tumor suppressor genes results in 5% LUAD and 5% LUSC inconclusive and, respectively, 2% and 1% misclassified. For large panels, 0%-1% was inconclusive and 1% misclassified for both LUAD and LUSC. Misclassifications occurred due to discordant KRAS mutations in clonal pairs or coincidentally shared PIK3CA mutations in non-clonal pairs.
Oncogene panels result in many inconclusive results, most of which can be resolved by adding tumor suppressor genes. Notwithstanding, 1%-2% of patients remain challenging. Large NGS panels detect mutations in more genes than the IASLC recommendations, allowing definitive clonality classification.
下一代测序（NGS）突变检测板已被广泛应用，常用于辅助对患有多个肿瘤的非小细胞肺癌（NSCLC）患者进行克隆性分类。然而，不同NGS检测板在克隆性分类方面的性能仍未明确。
我们从TRACERx421队列的肺腺癌（LUAD）和肺鳞状细胞癌（LUSC）中收集了210对原发性和转移性肿瘤样本，通过全外显子测序确定了其克隆性的金标准。我们使用了包含12至523个基因的四个NGS检测板，根据2024年国际肺癌研究协会（IASLC）的建议来确定克隆性。
使用癌基因检测板时，30%的LUAD和74%的LUSC样本对结果仍不明确，错误分类率分别为2%和0%。添加肿瘤抑制基因后，LUAD和LUSC的不确定率分别为5%和5%，错误分类率分别为2%和1%。对于大型检测板，LUAD和LUSC的不确定率均为0%-1%，错误分类率为1%。错误分类是由于克隆样本对中KRAS突变不一致或非克隆样本对中偶然共享PIK3CA突变所致。
癌基因检测板会产生许多不确定结果，其中大部分可通过添加肿瘤抑制基因来解决。尽管如此，仍有1%-2%的患者情况具有挑战性。大型NGS检测板检测到的基因突变比IASLC建议的更多，能够进行明确的克隆性分类。
