Mutation profile analysis of circulating tumor DNA in advanced lung cancer using targeted sequencing_West China Medical Journal

Authors：

BAI Ling , SONG Jiajia , ZHAI Jianzhao , LI Jin , LU Xiaojun , ZHOU Yi , ZHOU Juan ,  CHEN Hao

Department of Laboratory Medicine, West China Hospital, Sichuan University; Clinical Laboratory Medicine Research Center, West China Hospital, Sichuan University; Sichuan Clinical Research Center for Laboratory Medicine; Institution of Medical and Engineering Integration for Molecular Diagnosis, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, P. R. China;

Corresponding?author：

CHEN Hao, Email: haochen@wchscu.edu.cn

Keywords：

Lung cancer; circulating tumor DNA; metastasis; targeted therapy

DOI：

10.7507/1002-0179.202508040

Video：

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Objective To analyze the genetic variation characteristics of circulating tumor DNA (ctDNA) in patients with advanced lung cancer, aiming to provide valuable information for clinical decision-making. Methods In this retrospective study, a total of 468 patients with advanced-stage lung cancer were enrolled at West China Hospital, Sichuan University, from February 2017 to November 2019. Peripheral blood from each patient was collected for next generation sequencing (NGS) based on Illumina NextSeq550 platform, using a 23-gene targeted panel. Results Totally, 74.57% of patients were found to have at least one genetic alteration in plasma ctDNA, and drug-sensitive or drug-resistance information was obtained in 68.19% of patients. Frequently mutated genes included EGFR (47.01%), ROS1 (7.91%), ALK (7.69%), and HER2 (6.41%). EGFR mutation was more common in non-smokers (P<0.05), while KRAS mutation tended to occur more frequently in males (P<0.01) and smokers (P<0.01). EGFR 19DEL (28.88%), T790M (25.07%), and L858R (23.43%) were the most common types of EGFR mutations, and some rare potential drug-resistant mutations like K806R, S768I and L861Q, were also detected. Patients with brain metastasis had a relatively lower positive rate (69.23%). EGFR mutations were associated with bone metastasis (P<0.001), PIK3CA and KRAS mutations were enriched in liver metastasis (P<0.01 and P<0.001, respectively), while FGFR1 and HER2 mutations were more common in adrenal metastasis (P<0.01 and P<0.05, respectively). Third-generation EGFR tyrosine kinase inhibitor significantly increased the detection rate of FGFR1 genetic alterations in patient ctDNA when administered as second-line therapy (P<0.01). Conclusions The mutation profile and positive rate of plasma ctDNA are closely associated with smoking history, sex and metastatic sites. Comprehensive plasma ctDNA profiling using NGS technology demonstrates clinical utility by revealing therapeutically sensitive or resistant alterations in over half of the advanced-stage lung cancer patients, thereby informing personalized treatment strategies.

Citation： BAI Ling, SONG Jiajia, ZHAI Jianzhao, LI Jin, LU Xiaojun, ZHOU Yi, ZHOU Juan, CHEN Hao. Mutation profile analysis of circulating tumor DNA in advanced lung cancer using targeted sequencing. West China Medical Journal, 2026, 41(4): 562-568. doi: 10.7507/1002-0179.202508040 Copy

1.	Bray F, Laversanne M, Sung H, et al. Global cancer statistics 2022: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin, 2024, 74(3): 229-263.
2.	Cappuzzo F, Ciuleanu T, Stelmakh L, et al. Erlotinib as maintenance treatment in advanced non-small-cell lung cancer: a multicentre, randomised, placebo-controlled phase 3 study. Lancet Oncol, 2010, 11(6): 521-529.
3.	J?nne PA, Yang JC, Kim DW, et al. AZD9291 in EGFR inhibitor-resistant non-small-cell lung cancer. N Engl J Med, 2015, 372(18): 1689-1699.
4.	Katayama R, Lovly CM, Shaw AT. Therapeutic targeting of anaplastic lymphoma kinase in lung cancer: a paradigm for precision cancer medicine. Clin Cancer Res, 2015, 21(10): 2227-2235.
5.	Lindeman NI, Cagle PT, Aisner DL, et al. Updated molecular testing guideline for the selection of lung cancer patients for treatment with targeted tyrosine kinase inhibitors: guideline from the College of American Pathologists, the International Association for the Study of Lung Cancer, and the Association for Molecular Pathology. Arch Pathol Lab Med, 2018, 142(3): 321-346.
6.	Esposito Abate R, Frezzetti D, Maiello MR, et al. Next generation sequencing-based profiling of cell free DNA in patients with advanced non-small cell lung cancer: advantages and pitfalls. Cancers (Basel), 2020, 12(12): 3804.
7.	Duffy MJ. Circulating tumor DNA (ctDNA) as a biomarker for lung cancer: early detection, monitoring and therapy prediction. Tumour Biol, 2024, 46(s1): S283-S295.
8.	Leighl NB, Page RD, Raymond VM, et al. Clinical utility of comprehensive cell-free DNA analysis to identify genomic biomarkers in patients with newly diagnosed metastatic non-small cell lung cancer. Clin Cancer Res, 2019, 25(15): 4691-4700.
9.	Odegaard JI, Vincent JJ, Mortimer S, et al. Validation of a plasma-based comprehensive cancer genotyping assay utilizing orthogonal tissue- and plasma-based methodologies. Clin Cancer Res, 2018, 24(15): 3539-3549.
10.	王磊, 劉先, 吳清輝, 等. 循環腫瘤 DNA 與 EGFR、KRAS、NRAS 突變型可切除非小細胞肺癌的病理關系. 局解手術學雜志, 2025, 34(2): 154-158.
11.	Aggarwal C, Thompson JC, Black TA, et al. Clinical implications of plasma-based genotyping with the delivery of personalized therapy in metastatic non-small cell lung cancer. JAMA Oncol, 2019, 5(2): 173-180.
12.	Mack PC, Banks KC, Espenschied CR, et al. Spectrum of driver mutations and clinical impact of circulating tumor DNA analysis in non-small cell lung cancer: analysis of over 8000 cases. Cancer, 2020, 126(14): 3219-3228.
13.	Chen H, Wang A, Wang J, et al. Target-based genomic profiling of ctDNA from Chinese non-small cell lung cancer patients: a result of real-world data. J Cancer Res Clin Oncol, 2020, 146(7): 1867-1876.
14.	Wen S, Dai L, Wang L, et al. Genomic signature of driver genes identified by target next-generation sequencing in Chinese non-small cell lung cancer. Oncologist, 2019, 24(11): e1070-e1081.
15.	Thompson JC, Yee SS, Troxel AB, et al. Detection of therapeutically targetable driver and resistance mutations in lung cancer patients by next-generation sequencing of cell-free circulating tumor DNA. Clin Cancer Res, 2016, 22(23): 5772-5782.
16.	Wood K, Hensing T, Malik R, et al. Prognostic and predictive value in KRAS in non-small-cell lung cancer: a review. JAMA Oncol, 2016, 2(6): 805-812.
17.	Ricciuti B, Son J, Okoro JJ, et al. Comparative analysis and isoform-specific therapeutic vulnerabilities of KRAS mutations in non-small cell lung cancer. Clin Cancer Res, 2022, 28(8): 1640-1650.
18.	Shi Y, Au JS, Thongprasert S, et al. A prospective, molecular epidemiology study of EGFR mutations in Asian patients with advanced non-small-cell lung cancer of adenocarcinoma histology (PIONEER). J Thorac Oncol, 2014, 9(2): 154-162.
19.	Chiu CH, Yang CT, Shih JY, et al. Epidermal growth factor receptor tyrosine kinase inhibitor treatment response in advanced lung adenocarcinomas with G719X/L861Q/S768I mutations. J Thorac Oncol, 2015, 10(5): 793-799.
20.	Leventakos K, Kipp BR, Rumilla KM, et al. S768I mutation in EGFR in patients with lung cancer. J Thorac Oncol, 2016, 11(10): 1798-1801.
21.	Piotrowska Z, Nagy R, Fairclough S, et al. OA 09.01 characterizing the genomic landscape of EGFR C797S in lung cancer using ctDNA next-generation sequencing. J Thorac Oncol, 2017, 12(11): S1767.
22.	Thress KS, Paweletz CP, Felip E, et al. Acquired EGFR C797S mutation mediates resistance to AZD9291 in non-small cell lung cancer harboring EGFR T790M. Nat Med, 2015, 21(6): 560-562.
23.	Yu HA, Tian SK, Drilon AE, et al. Acquired resistance of EGFR-mutant lung cancer to a T790M-specific EGFR inhibitor: emergence of a third mutation (C797S) in the EGFR tyrosine kinase domain. JAMA Oncol, 2015, 1(7): 982-984.
24.	Leonetti A, Sharma S, Minari R, et al. Resistance mechanisms to osimertinib in EGFR-mutated non-small cell lung cancer. Br J Cancer, 2019, 121(9): 725-737.
25.	Nguyen B, Fong C, Luthra A, et al. Genomic characterization of metastatic patterns from prospective clinical sequencing of 25, 000 patients. Cell, 2022, 185(3): 563-575. e11.
26.	Testa U, Castelli G, Pelosi E. Lung cancers: molecular characterization, clonal heterogeneity and evolution, and cancer stem cells. Cancers (Basel), 2018, 10(8): 248.
27.	Cescon DW, Bratman SV, Chan SM, et al. Circulating tumor DNA and liquid biopsy in oncology. Nat Cancer, 2020, 1(3): 276-290.
28.	Supplee JG, Milan MSD, Lim LP, et al. Sensitivity of next-generation sequencing assays detecting oncogenic fusions in plasma cell-free DNA. Lung Cancer, 2019, 134: 96-99.
29.	Aggarwal C, Rolfo CD, Oxnard GR, et al. Strategies for the successful implementation of plasma-based NSCLC genotyping in clinical practice. Nat Rev Clin Oncol, 2021, 18(1): 56-62.
30.	雷思雨, 王燕. 循環腫瘤 DNA 在非小細胞肺癌診療中的研究進展. 中國肺癌雜志, 2022, 25(9): 665-670.

1. Bray F, Laversanne M, Sung H, et al. Global cancer statistics 2022: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin, 2024, 74(3): 229-263.
2. Cappuzzo F, Ciuleanu T, Stelmakh L, et al. Erlotinib as maintenance treatment in advanced non-small-cell lung cancer: a multicentre, randomised, placebo-controlled phase 3 study. Lancet Oncol, 2010, 11(6): 521-529.
3. J?nne PA, Yang JC, Kim DW, et al. AZD9291 in EGFR inhibitor-resistant non-small-cell lung cancer. N Engl J Med, 2015, 372(18): 1689-1699.
4. Katayama R, Lovly CM, Shaw AT. Therapeutic targeting of anaplastic lymphoma kinase in lung cancer: a paradigm for precision cancer medicine. Clin Cancer Res, 2015, 21(10): 2227-2235.
5. Lindeman NI, Cagle PT, Aisner DL, et al. Updated molecular testing guideline for the selection of lung cancer patients for treatment with targeted tyrosine kinase inhibitors: guideline from the College of American Pathologists, the International Association for the Study of Lung Cancer, and the Association for Molecular Pathology. Arch Pathol Lab Med, 2018, 142(3): 321-346.
6. Esposito Abate R, Frezzetti D, Maiello MR, et al. Next generation sequencing-based profiling of cell free DNA in patients with advanced non-small cell lung cancer: advantages and pitfalls. Cancers (Basel), 2020, 12(12): 3804.
7. Duffy MJ. Circulating tumor DNA (ctDNA) as a biomarker for lung cancer: early detection, monitoring and therapy prediction. Tumour Biol, 2024, 46(s1): S283-S295.
8. Leighl NB, Page RD, Raymond VM, et al. Clinical utility of comprehensive cell-free DNA analysis to identify genomic biomarkers in patients with newly diagnosed metastatic non-small cell lung cancer. Clin Cancer Res, 2019, 25(15): 4691-4700.
9. Odegaard JI, Vincent JJ, Mortimer S, et al. Validation of a plasma-based comprehensive cancer genotyping assay utilizing orthogonal tissue- and plasma-based methodologies. Clin Cancer Res, 2018, 24(15): 3539-3549.
10. 王磊, 劉先, 吳清輝, 等. 循環腫瘤 DNA 與 EGFR、KRAS、NRAS 突變型可切除非小細胞肺癌的病理關系. 局解手術學雜志, 2025, 34(2): 154-158.
11. Aggarwal C, Thompson JC, Black TA, et al. Clinical implications of plasma-based genotyping with the delivery of personalized therapy in metastatic non-small cell lung cancer. JAMA Oncol, 2019, 5(2): 173-180.
12. Mack PC, Banks KC, Espenschied CR, et al. Spectrum of driver mutations and clinical impact of circulating tumor DNA analysis in non-small cell lung cancer: analysis of over 8000 cases. Cancer, 2020, 126(14): 3219-3228.
13. Chen H, Wang A, Wang J, et al. Target-based genomic profiling of ctDNA from Chinese non-small cell lung cancer patients: a result of real-world data. J Cancer Res Clin Oncol, 2020, 146(7): 1867-1876.
14. Wen S, Dai L, Wang L, et al. Genomic signature of driver genes identified by target next-generation sequencing in Chinese non-small cell lung cancer. Oncologist, 2019, 24(11): e1070-e1081.
15. Thompson JC, Yee SS, Troxel AB, et al. Detection of therapeutically targetable driver and resistance mutations in lung cancer patients by next-generation sequencing of cell-free circulating tumor DNA. Clin Cancer Res, 2016, 22(23): 5772-5782.
16. Wood K, Hensing T, Malik R, et al. Prognostic and predictive value in KRAS in non-small-cell lung cancer: a review. JAMA Oncol, 2016, 2(6): 805-812.
17. Ricciuti B, Son J, Okoro JJ, et al. Comparative analysis and isoform-specific therapeutic vulnerabilities of KRAS mutations in non-small cell lung cancer. Clin Cancer Res, 2022, 28(8): 1640-1650.
18. Shi Y, Au JS, Thongprasert S, et al. A prospective, molecular epidemiology study of EGFR mutations in Asian patients with advanced non-small-cell lung cancer of adenocarcinoma histology (PIONEER). J Thorac Oncol, 2014, 9(2): 154-162.
19. Chiu CH, Yang CT, Shih JY, et al. Epidermal growth factor receptor tyrosine kinase inhibitor treatment response in advanced lung adenocarcinomas with G719X/L861Q/S768I mutations. J Thorac Oncol, 2015, 10(5): 793-799.
20. Leventakos K, Kipp BR, Rumilla KM, et al. S768I mutation in EGFR in patients with lung cancer. J Thorac Oncol, 2016, 11(10): 1798-1801.
21. Piotrowska Z, Nagy R, Fairclough S, et al. OA 09.01 characterizing the genomic landscape of EGFR C797S in lung cancer using ctDNA next-generation sequencing. J Thorac Oncol, 2017, 12(11): S1767.
22. Thress KS, Paweletz CP, Felip E, et al. Acquired EGFR C797S mutation mediates resistance to AZD9291 in non-small cell lung cancer harboring EGFR T790M. Nat Med, 2015, 21(6): 560-562.
23. Yu HA, Tian SK, Drilon AE, et al. Acquired resistance of EGFR-mutant lung cancer to a T790M-specific EGFR inhibitor: emergence of a third mutation (C797S) in the EGFR tyrosine kinase domain. JAMA Oncol, 2015, 1(7): 982-984.
24. Leonetti A, Sharma S, Minari R, et al. Resistance mechanisms to osimertinib in EGFR-mutated non-small cell lung cancer. Br J Cancer, 2019, 121(9): 725-737.
25. Nguyen B, Fong C, Luthra A, et al. Genomic characterization of metastatic patterns from prospective clinical sequencing of 25, 000 patients. Cell, 2022, 185(3): 563-575. e11.
26. Testa U, Castelli G, Pelosi E. Lung cancers: molecular characterization, clonal heterogeneity and evolution, and cancer stem cells. Cancers (Basel), 2018, 10(8): 248.
27. Cescon DW, Bratman SV, Chan SM, et al. Circulating tumor DNA and liquid biopsy in oncology. Nat Cancer, 2020, 1(3): 276-290.
28. Supplee JG, Milan MSD, Lim LP, et al. Sensitivity of next-generation sequencing assays detecting oncogenic fusions in plasma cell-free DNA. Lung Cancer, 2019, 134: 96-99.
29. Aggarwal C, Rolfo CD, Oxnard GR, et al. Strategies for the successful implementation of plasma-based NSCLC genotyping in clinical practice. Nat Rev Clin Oncol, 2021, 18(1): 56-62.
30. 雷思雨, 王燕. 循環腫瘤 DNA 在非小細胞肺癌診療中的研究進展. 中國肺癌雜志, 2022, 25(9): 665-670.

West China Medical Journal

Mutation profile analysis of circulating tumor DNA in advanced lung cancer using targeted sequencing

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