Urinary circulating DNA detection for dynamic tracking of EGFR mutations for NSCLC patients treated with EGFR-TKIs
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Changes in EGFR profiles of non small cell lung cancer (NSCLC) patients correlates to clinical outcome. Extracting quality tumor tissue remains a challenge for molecular profiling. Our study aims to ascertain the clinical relevance of urinary cell free DNA as an alternative tumor material source.
150 patients with activating EGFR mutation and received EGFR-TKIs were recruited to participate in the serial monitoring study. Matched primary tumor samples were taken together with blood and urine specimens before the initiation of TKIs. The EGFR mutation testing was performed and quantified using ddPCR. For serial time point measurements, urine and blood samples were extracted at 1-month intervals for duration of 9 months.
Urinary ctDNA yielded a close agreement of 88 % on EGFR mutation status when compared to primary tissue at baseline. Almost all samples detected via urine specimens were uncovered in plasma samples. Analysis of urinary cell free DNA at different time points showed a strong correlation to treatment efficacy. Interestingly, a secondary EGFR mutation T790M was detected for 53 % of the patients during monitoring. The results were corroborated with the plasma ctDNA analysis. The T790M+ group had a reduced median survival when compared to the wildtype group.
Urinary cell free DNA may be a potential alternative to conventional primary tissue based EGFR mutation testing. Our findings showed that the assay sensitivity was comparable to results from blood plasma. Urinary samples being noninvasive and readily available have clinical utility for monitoring of EGFR TKI treatment.
KeywordsNSCLC Cell free DNA TKI treatment Drug resistance Liquid biopsy EGFR mutations
This work was supported by a research grant provided by the Xiangyang Central Hospital.
Compliance with ethical standards
Conflict of interest
All authors declare no conflict of interest.
All human and animal studies have been approved by the appropriate ethics committee, and have therefore been performed in accordance with the ethical standards laid down in the 1964 Declaration of Helsinki and its later amendments.
- 2.Zhou C, Wu Y-L, Chen G, Feng J, Liu X-Q, Wang C, et al. Erlotinib versus chemotherapy as first-line treatment for patients with advanced EGFR mutation-positive non-small-cell lung cancer (OPTIMAL, CTONG-0802): a multicentre, open-label, randomised, phase 3 study. Lancet Oncol. 2011;12(8):735–42. doi: 10.1016/S1470-2045(11)70184-X.CrossRefPubMedGoogle Scholar
- 4.Yu HA, Arcila ME, Rekhtman N, Sima CS, Zakowski MF, Pao W, et al. Analysis of tumor specimens at the time of acquired resistance to EGFR-TKI therapy in 155 patients with EGFR-mutant lung cancers. Am Assoc Cancer Res. 2013;19(8):2240–7.Google Scholar
- 5.Chouaid C, Dujon C, Do P, Monnet I, Madroszyk A, Le Caer H, et al. Feasibility and clinical impact of re-biopsy in advanced non small-cell lung cancer: a prospective multicenter study in a real-world setting (GFPC study 12-01). Lung Cancer. 2014;86(2):170–3. doi: 10.1016/j.lungcan.2014.08.016.CrossRefPubMedGoogle Scholar
- 8.Douillard J-Y, Ostoros G, Cobo M, Ciuleanu T, Cole R, McWalter G, et al. Gefitinib treatment in EGFR mutated caucasian NSCLC: circulating-free tumor DNA as a surrogate for determination of EGFR status. J Thorac Oncol. 2014;9(9):1345–53. doi: 10.1097/JTO.0000000000000263.CrossRefPubMedPubMedCentralGoogle Scholar
- 9.Oxnard GR, Paweletz CP, Kuang Y, Mach SL, O’Connell A, Messineo MM, et al. Noninvasive detection of response and resistance in EGFR-mutant lung cancer using quantitative next-generation genotyping of cell-free plasma DNA. Clin Cancer Res. 2014;20(6):1698–705.CrossRefPubMedPubMedCentralGoogle Scholar
- 13.Fischbach FT, Dunning MB. A manual of laboratory and diagnostic tests. Philadelphia: Lippincott Williams & Wilkins; 2009.Google Scholar
- 17.Diehl F, Schmidt K, Choti MA, Romans K, Goodman S, Li M et al. Circulating mutant DNA to assess tumor dynamics. Nat Med. 2008;14(9):985–990. http://www.nature.com/nm/journal/v14/n9/suppinfo/nm.1789_S1.html.
- 18.Szpechcinski A, Chorostowska-Wynimko J, Struniawski R, Kupis W, Rudzinski P, Langfort R, et al. Cell-free DNA levels in plasma of patients with non-small-cell lung cancer and inflammatory lung disease. Br J Cancer. 2015;113(3):476–83. doi: 10.1038/bjc.2015.225.CrossRefPubMedPubMedCentralGoogle Scholar
- 21.Utting M, Werner W, Dahse R, Schubert J, Junker K. Microsatellite analysis of free tumor DNA in urine, serum, and plasma of patients. Am Assoc Cancer Res. 2002;8(1):35–40.Google Scholar
- 23.Lynch TJ, Bell DW, Sordella R, Gurubhagavatula S, Okimoto RA, Brannigan BW, et al. Activating mutations in the epidermal growth factor receptor underlying responsiveness of non-small-cell lung cancer to gefitinib. N Engl J Med. 2004;350(21):2129–39. doi: 10.1056/NEJMoa040938.CrossRefPubMedGoogle Scholar
- 24.Pao W, Miller VA, Politi KA, Riely GJ, Somwar R, Zakowski MF, et al. Acquired resistance of lung adenocarcinomas to gefitinib or erlotinib is associated with a second mutation in the EGFR kinase domain. PLoS Med. 2005;2(3):e73. doi: 10.1371/journal.pmed.0020073.CrossRefPubMedPubMedCentralGoogle Scholar
- 26.Zheng D, Ye X, Zhang MZ, Sun Y, Wang JY, Ni J et al. Plasma EGFR T790M ctDNA status is associated with clinical outcome in advanced NSCLC patients with acquired EGFR-TKI resistance. Sci Rep. 2016;6:20913. doi: 10.1038/srep20913. http://www.nature.com/articles/srep20913-supplementary-information.
- 28.Lee Y, Lee GK, Lee Y-S, Zhang W, Hwang J-A, Nam B-H, et al. Clinical outcome according to the level of preexisting epidermal growth factor receptor T790M mutation in patients with lung cancer harboring sensitive epidermal growth factor receptor mutations. Cancer. 2014;120(14):2090–8. doi: 10.1002/cncr.28711.CrossRefPubMedGoogle Scholar