Quantitative analysis of cell-free DNA by droplet digital PCR reveals the presence of EGFR mutations in non-malignant lung pathologies

Submitted: December 24, 2020
Accepted: April 18, 2021
Published: May 4, 2021
Abstract Views: 1451
PDF: 533
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Tyrosine kinase inhibitors (TKIs) targeting epidermal growth factor receptor (EGFR) are effectively used in treatment of non-small cell lung cancer (NSCLC). Mutation profile of tyrosine kinase domain of EGFR determines the eligibility of the patients for tyrosine kinase inhibitor (TKI) therapy. Liquid biopsy, which relies on circulating tumor-derived nucleic acids, has emerged as an effective tool in lung cancer management with proven diagnostic, prognostic and predictive applications. We screened 100 subjects, suspected to have lung malignancy, for four hotspot mutations including three activating (G719S, Ex19del E746-A750 and L858R) and one acquired (T790M, de novo) in EGFR gene by droplet digital PCR (ddPCR). While 97 subjects were subsequently confirmed to have lung malignancy based on histo/cytopathological studies, three cases turned out to be non-malignant lung pathologies that were completely cured by antibiotic therapy. Intriguingly, ddPCR revealed the presence of EGFR mutations in these non-malignant subjects. Two cases showed the presence of G719S and T790M mutations respectively and another had compound mutations (T790M and L858R). The detection of EGFR mutations in non-malignant pulmonary conditions opens up a new area of research.



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Pao W, Girard N. New driver mutations in non-small-cell lung cancer. Lancet Oncol 2011;12:175-80. DOI: https://doi.org/10.1016/S1470-2045(10)70087-5
Ma J, Li N, Guarnera M, Jiang F. Quantification of plasma miRNAs by digital PCR for cancer diagnosis. Biomark Insights 2013;8:127-36. DOI: https://doi.org/10.4137/BMI.S13154
Shigematsu H, Lin L, Takahashi T, et al. Clinical and biological features associated with epidermal growth factor receptor gene mutations in lung cancers. J Natnl Cancer Inst 2005;97:339-46. DOI: https://doi.org/10.1093/jnci/dji055
Forshew T, Murtaza M, Parkinson C, et al. Non-invasive identification and monitoring of cancer mutations by targeted deep sequencing of plasma DNA. Sci Transl Med 2012;4:136ra68. DOI: https://doi.org/10.1126/scitranslmed.3003726
Oxnard G, Paweletz C, Kuang Y, 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;20:1698-705. DOI: https://doi.org/10.1158/1078-0432.CCR-13-2482
Kobayashi S, Boggon TJ, Dayaram T, et al. EGFR mutation and resistance of non-small cell lung cancer to gefitinib. N Engl J Med 2005;352:786-92. DOI: https://doi.org/10.1056/NEJMoa044238
Sharma SV, Bell DW, Settleman J, Haber DA. Epidermal growth factor receptor mutations in lung cancer. Nat Rev Cancer 2007;7:169-81. DOI: https://doi.org/10.1038/nrc2088
Li K, Yang M, Liang N, Li S. Determining EGFR-TKI sensitivity of G719X and other uncommon EGFR mutations in non-small cell lung cancer: Perplexity and solution (Review). Oncol Rep 2017;37:1347-58. DOI: https://doi.org/10.3892/or.2017.5409
Crowley E, Nicolantonio F, Loupakis F, Bardelli A. Liquid biopsy: monitoring cancer-genetics in the blood. Nat Rev Clin Oncol 2013;10:472-84. DOI: https://doi.org/10.1038/nrclinonc.2013.110
Santarpia M, Liguori A, D’Aveni A, et al. Liquid biopsy for lung cancer early detection. J Thorac Dis 2018;10:S882-97. DOI: https://doi.org/10.21037/jtd.2018.03.81
Zhang R, Chen B, Tong X, et al. Diagnostic accuracy of droplet digital PCR for detection of EGFR T790M mutation in circulating tumor DNA. Cancer Manag Res 2018;10:1209-18. DOI: https://doi.org/10.2147/CMAR.S161382
Watanabe M, Kawaguchi T, Isa S, et al. Ultra-sensitive detection of the pre-treatment EGFR T790M mutation in non-small cell lung cancer patients with an EGFR-activating mutation using droplet digital PCR. Clin Cancer Res 2015;21:3552-60. DOI: https://doi.org/10.1158/1078-0432.CCR-14-2151
Zhang B, Xu C, Shao Y, et al. Comparison of droplet digital PCR and conventional quantitative PCR for measuring EGFR gene mutation. Exp Ther Med 2015;9:1383-8. DOI: https://doi.org/10.3892/etm.2015.2221
Hindson B, Ness K, Masquelier D, et al. High-throughput droplet digital PCR system for absolute quantitation of DNA copy number. Anal Chem 2011;83:8604-10. DOI: https://doi.org/10.1021/ac202028g
Hwang IK, Paik SS, Lee SH. Impact of pulmonary tuberculosis on the EGFR mutational status and clinical outcome in patients with lung adenocarcinoma. Cancer Res Treat 2019;51:158-68. DOI: https://doi.org/10.4143/crt.2018.084
Luo YH, Wu CH, Wu WS, et al. Association between tumor epidermal growth factor receptor mutation and pulmonary tuberculosis in patients with adenocarcinoma of the lungs. J Thorac Oncol 2012;7:299-05. DOI: https://doi.org/10.1097/JTO.0b013e31823c588d
Bell DW, Gore I, Okimoto RA, et al. Inherited susceptibility to lung cancer may be associated with the T790M drug resistance mutation in EGFR. Nat Genet 2005;37:1315-6. DOI: https://doi.org/10.1038/ng1671
Girard N, Lou E, Azzoli CG, et al. Analysis of genetic variants in never-smokers with lung cancer facilitated by an internet-based blood collection protocol: A preliminary report. Clin Cancer Res 2010;16:755-63. DOI: https://doi.org/10.1158/1078-0432.CCR-09-2437
Yu HA, Arcila ME, Harlan M, et al. Germline EGFR T790M mutation found in multiple members of a familial cohort. J Thorac Oncol 2014;9:554-8. DOI: https://doi.org/10.1097/JTO.0000000000000052
Lou Y, Pecot CV, Tran HT, et al. Germline mutation of T790M and dual/multiple EGFR mutations in patients with lung adenocarcinoma. Clin Lung Cancer 2016;17:e5-11. DOI: https://doi.org/10.1016/j.cllc.2015.11.003
Nambiar M., Raghavan S.C. Prevalence and analysis of t(14;18) and t(11;14) chromosomal translocations in healthy Indian population. Ann Hematol 2010;89:35-43. DOI: https://doi.org/10.1007/s00277-009-0755-1
Chang JC, Montecalvo J, Borsu L, et al. Bronchiolar adenoma: Expansion of the concept of ciliated muconodular papillary tumors with proposal for revised terminology based on morphologic, immunophenotypic, and genomic analysis of 25 cases. Am J Surg Pathol 2018;42:1010-26. DOI: https://doi.org/10.1097/PAS.0000000000001086

How to Cite

Venkataram, Rajesh, Srividya Arjuna, Giridhar Belur Hosmane, and Anirban Chakraborty. 2021. “Quantitative Analysis of Cell-Free DNA by Droplet Digital PCR Reveals the Presence of EGFR Mutations in Non-Malignant Lung Pathologies”. Monaldi Archives for Chest Disease 91 (3). https://doi.org/10.4081/monaldi.2021.1748.