中国肺癌杂志2026，Vol.29Issue(7)：491-499,9.DOI:10.3779/j.issn.1009-3419.2026.106.18
基于重复NGS检测的非小细胞肺癌驱动基因状态变化与稳定性分级
State Changes and Stability Grading of Driver Genes in Non-small Cell Lung Cancer Based on Repeated NGS Testing
摘要
Abstract
Background and objective Next-generation sequencing(NGS)-based driver gene testing has become a routine component of molecular subtyping and precision therapy for non-small cell lung cancer(NSCLC).Dynamic genomic monitoring facili-tates early detection of resistance-related molecular alterations and informs timely therapeutic adjustments.However,standardized criteria for evaluating the stability of serial NGS testing are currently lacking,and the applicability of NGS using formalin-fixed paraffin-embedded(FFPE)specimens for dynamic monitoring remains poorly defined.This study aims to establish a stability grading system for driver gene status alterations based on repeated NGS testing,and to provide evidence-based support for clinical repeat biopsy strategies.Methods Data from 1232 patients with NSCLC who underwent two or more NGS tests on FFPE tissue specimens at Beijing Chest Hospital be-tween June 2019 and April 2026 were collected retrospectively.Patients with an interval of≥4 months between the initial and last tests were included to ensure the representativeness of temporal analysis,resulting in a main analysis cohort of 942 patients.The Kappa consistency test was used to evaluate the state stability of nine core driver genes[epidermal growth factor receptor(EGFR),Kirsten rat sarcoma viral oncogene homolog(KRAS),anaplastic lymphoma kinase(ALK),ROS proto-oncogene 1,receptor tyrosine kinase(ROS1),mesenchy-mal-epithelial transition factor(MET),rearranged during transfection(RET),v-raf murine sarcoma viral oncogene homolog B1(BRAF),erb-b2 receptor tyrosine kinase 2(ERBB2),and phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha(PIK3CA)]and to construct a five-level grading system.Paired variant allele frequency(VAF)differences were compared using the Wilcoxon signed-rank test.Independent influencing factors for mutation accumulation were identified by binary Logistic regression.Results The state stability of the nine genes was classified into five levels:EGFR showed high stability(Kappa=0.838),ROS1/ALK/KRAS good stability,BRAF/PIK3CA/RET moderate stability,and ERBB2 low stability,and MET showed high instability.MET exhibited the highest rate of state change(9.3％)with a raw observed agreement of 90.7％.Its Kappa value(0.172)was influenced by the low prevalence(3.7％)compression effect and should therefore be interpreted alongside the observed agreement(90.7％)and the prevalence-adjusted and bias-adjusted Kappa(PABAK).The VAF of PIK3CA increased significantly(P=0.005).T790M positivity increased from 5.8％to 10.8％,and 30 new C797S mutations were detected at the last test(13 with T790M,17 without).The overall rate of new driver gene variants in the main cohort was 18.0％.Bi-nary Logistic regression showed that a lower number of initial mutated genes was the only independent predictor of new variants[odds ratio(OR)=0.399,P<0.001],while sex and detection interval showed no independent association.Conclusion A five level stability grading system for state changes of driver genes in NSCLC based on repeated NGS testing has been established.MET showed the most frequent state changes,which should be interpreted in conjunction with the prevalence effect.The VAF increase of PIK3CA is an observational find-ing,and its clinical significance requires further prospective validation.A lower initial mutation burden may reflect tumor clonal complexity and was associated with a higher likelihood of subsequent acquisition of new variants.FFPE based NGS is applicable for repeated testing at clinical treatment decision nodes.关键词
肺肿瘤/驱动基因/下一代测序/基因状态稳定性/纵向研究Key words
Lung neoplasms/Driver genes/Next-generation sequencing/Gene status stability/Longitudinal studies引用本文复制引用
赵丹,穆晶,车南颖,徐福东,张丽丽,张娜娜,刘子臣,李琨,张晨,周立娟,董宇杰..基于重复NGS检测的非小细胞肺癌驱动基因状态变化与稳定性分级[J].中国肺癌杂志,2026,29(7):491-499,9.基金项目
本研究受国家自然科学基金项目(No.82472378)、北京市属医学科研院所公益发展改革试点项目(京医研2023-15)、通州区高层次人才发展支持计划(No.YH201901)和北京市医院管理中"心登峰"人才培养计划(No.DFL20241601)资助 This study was supported by the grants from National Natural Science Foundation of China(No.82472378,to Nanying CHE),Pilot Project for Public Welfare Development Reform of Beijing Municipal Medical Research Institutions(Jing Yi Yan 2023-15,to Nany-ing CHE),Tongzhou District High-level Talent Development Program(No.YH201901,to Nanying CHE)and Beijing Hospital Authority"Dengfeng"Talent Training Program(No.DFL20241601,to Nanying CHE). （No.82472378）
