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EGFR Mutation Analysis in Non-small Cell Lung Carcinoma Patients: A Liquid Biopsy Approach

  • Jigna Joshi
  • Apexa Raval
  • Urja Desai
  • Vinal Upadhyay
  • Mansi Bhavsar
  • Kanisha Shah
  • Rakesh Rawal
  • Harsha Panchal
  • Franky ShahEmail author
Original Research Article
  • 26 Downloads

Abstract

In the era of the targeted therapy identification of EGFR mutation detection in lung cancer is extremely helpful to predict the treatment efficacy of EGFR tyrosine kinase inhibitors (TKIs). Unfortunately, the inadequacy and quality of the biopsy samples are the major obstacles in molecular testing of EGFR mutation in lung cancer. To address this issue, the present study intended to use liquid biopsy as the non-invasive method for EGFR mutation detection. A total of 31 patients with an advanced stage of lung cancer were enrolled in the study from which cell-free DNA (cfDNA) and FFPE tissue DNA was extracted. Extracted DNA samples were analyzed for further EGFR exon specific mutation analysis by ARMS-PCR. Data were analyzed statistically using SPSS software. In cfDNA samples, the prevalence of wild type EGFR was 48% while the prevalence of TKI resistant and TKI sensitive mutations were 3%. Conversely, in tissue DNA samples, the prevalence of wild type, TKI sensitive and TKI resistant mutations were 48%, 19%, and 3%, respectively. The overall concordance of EGFR mutation between cfDNA and tissue DNA was 83%. McNemar’s test revealed that there was no significant difference between EGFR expression of cfDNA and tissue DNA samples. Additionally, the significant-high incidence of TKI resistant mutations was observed in tobacco habituates, indicating the role of carcinogens present in the tobacco in developing resistant mutations. In conclusion, our data suggest that evaluation of EGFR mutation from cfDNA samples is practicable as a non-invasive tool in patients with advanced-stage of lung cancer.

Keywords

ARMS real-time PCR Cell-free DNA EGFR mutation Non-small cell lung cancer 

Notes

Acknowledgements

The study has been financially supported by The Gujarat Cancer & Research Institute/The Gujarat Cancer Society.

Funding

The Gujarat Cancer Society/The Gujarat Cancer & Research Institute (RE/74/MO/14).

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional review committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

Informed consent

Informed consent was obtained from all individual participants included in the study.

References

  1. 1.
    Ferlay J, Shin HR, Bray F, Forman D, Mathers C, Parkin DM. Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008. Int J Cancer. 2010;127:2893–917.CrossRefGoogle Scholar
  2. 2.
    Ferlay J, Soerjomataram I, Ervik M, Eser S, Mathers C, Rebelo M, et al. Cancer incidence and mortality worldwide: sources, methods and major patterns in GLOBOCAN 2012. Int J Cancer. 2015;1(136):E359–86.CrossRefGoogle Scholar
  3. 3.
    Malik PS, Raina V. Lung cancer: prevalent trends and emerging concepts. Indian J Med Res. 2015;141:5–7.CrossRefGoogle Scholar
  4. 4.
    Chung C. Tyrosine kinase inhibitors for epidermal growth factor receptor gene mutation–positive non-small cell lung cancers: an update for recent advances in therapeutics. J Oncol Pharm Pract. 2016;22:461–76.CrossRefGoogle Scholar
  5. 5.
    Bethune G, Bethune D, Ridgway N, Xu Z. Epidermal growth factor receptor (EGFR) in lung cancer: an overview and update. J Thorac Dis. 2010;2:48–51.PubMedPubMedCentralGoogle Scholar
  6. 6.
    Gazdar AF. Activating and resistance mutations of EGFR in non-small-cell lung cancer: role in clinical response to EGFR tyrosine kinase inhibitors. Oncogene. 2009;28:S24–31.CrossRefGoogle Scholar
  7. 7.
    Inamura K, Ninomiya H, Ishikawa Y, Matsubara O. Is the epidermal growth factor receptor status in lung cancers reflected in clinicopathologic features? Arch Pathol Lab Med. 2010;134:66–72.PubMedGoogle Scholar
  8. 8.
    Ohsaki Y, Tanno S, Fujita Y, Toyoshima E, Fujiuchi S, Nishigaki Y, et al. Epidermal growth factor receptor expression correlates with poor prognosis in non-small cell lung cancer patients with p53 overexpression. Oncol Rep. 2000;7:603–7.PubMedGoogle Scholar
  9. 9.
    Marchetti A, Martella C, Felicioni L, Barassi F, Salvatore S, Chella A, et al. EGFR mutations in non–small-cell lung cancer: analysis of a large series of cases and development of a rapid and sensitive method for diagnostic screening with potential implications on pharmacologic treatment. J Clin Oncol. 2005;23:857–65.CrossRefGoogle Scholar
  10. 10.
    Bartholomew C, Eastlake L, Dunn P, Yiannakis D. EGFR targeted therapy in lung cancer; an evolving story. Respir Med Case Rep. 2017;20:137–40.PubMedPubMedCentralGoogle Scholar
  11. 11.
    Douillard JY, 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;9:1345–53.CrossRefGoogle Scholar
  12. 12.
    Oxnard GR, Thress KS, Alden RS, Lawrance R, Paweletz CP, Cantarini M, et al. Association between plasma genotyping and outcomes of treatment with Osimertinib (AZD9291) in advanced non-small-cell lung cancer. J Clin Oncol. 2016;34:3375–82.CrossRefGoogle Scholar
  13. 13.
    O’Neill AC, Jagannathan JP, Ramaiya NH. Evolving cancer classification in the era of personalized medicine: a primer for radiologists. Korean J Radiol. 2017;18:6–17.CrossRefGoogle Scholar
  14. 14.
    Tsao MS, Sakurada A, Cutz JC, Zhu CQ, Kamel-Reid S, Squire J, et al. Erlotinib in lung cancer-molecular and clinical predictors of outcome. N Engl J Med. 2005;353:133–44.CrossRefGoogle Scholar
  15. 15.
    Kimura H, Suminoe M, Kasahara K, Sone T, Araya T, Tamori S, et al. Evaluation of epidermal growth factor receptor mutation status in serum DNA as a predictor of response to gefitinib (IRESSA). Br J Cancer. 2007;97:778–84.CrossRefGoogle Scholar
  16. 16.
    Vallee A, Marcq M, Bizieux A, Kouri CE, Lacroix H, Bennouna J, et al. Plasma is a better source of tumor-derived circulating cell-free DNA than serum for the detection of EGFR alterations in lung tumor patients. Lung Cancer. 2013;82:373–4.CrossRefGoogle Scholar
  17. 17.
    Nishino M, Jackman DM, Hatabu H, Jänne PA, Johnson BE, Van den Abbeele AD. Imaging of lung cancer in the era of molecular medicine. Acad Radiol. 2011;18:424–36.CrossRefGoogle Scholar
  18. 18.
    Ishii H, Azuma K, Sakai K, Kawahara A, Yamada K, Tokito T, et al. Digital PCR analysis of plasma cell-free DNA for non-invasive detection of drug resistance mechanisms in EGFR mutant NSCLC: correlation with paired tumor samples. Oncotarget. 2015;6:30850–8.CrossRefGoogle Scholar
  19. 19.
    Seki Y, Fujiwara Y, Kohno T, Takai E, Sunami K, Goto Y, et al. Picoliter-droplet digital polymerase chain reaction-based analysis of cell-free plasma DNA to assess EGFR mutations in lung adenocarcinoma that confer resistance to tyrosine-kinase inhibitors. Oncologist. 2016;21:156–64.CrossRefGoogle Scholar
  20. 20.
    Lee JY, Qing X, Xiumin W, Yali B, Chi S, Bak SH, et al. Longitudinal monitoring of EGFR mutations in plasma predicts outcomes of NSCLC patients treated with EGFR TKIs: Korean lung cancer consortium (KLCC-12-02). Oncotarget. 2016;7:6984–93.PubMedPubMedCentralGoogle Scholar
  21. 21.
    Jahr S, Hentze H, Englisch S, Hardt D, Fackelmayer FO, Hesch RD, et al. DNA fragments in the blood plasma of cancer patients: quantitations and evidence for their origin from apoptotic and necrotic cells. Cancer Res. 2001;61:1659–65.PubMedGoogle Scholar
  22. 22.
    Bai H, Mao L, Wang HS, Zhao J, Yang L, An TT, et al. Epidermal growth factor receptor mutations in plasma DNA samples predict tumor response in Chinese patients with stages IIIB to IV non-small-cell lung cancer. J Clin Oncol. 2009;27:2653–9.CrossRefGoogle Scholar
  23. 23.
    Kim HR, Lee SY, Hyun DS, Lee MK, Lee HK, Choi CM, et al. Detection of EGFR mutations in circulating free DNA by PNA-mediated PCR clamping. J Exp Clin Cancer Res. 2013;32:50.CrossRefGoogle Scholar
  24. 24.
    Couraud S, Vaca-Paniagua F, Villar S, Oliver J, Schuster T, Blanché H, et al. Noninvasive diagnosis of actionable mutations by deep sequencing of circulating free DNA in lung cancer from never-smokers: a proof-of-concept study from BioCAST/IFCT-1002. Clin Cancer Res. 2014;20:4613–24.CrossRefGoogle Scholar
  25. 25.
    Wang Y, Duan J, Chen H, Bai H, An T, Zhao J, et al. Analysis of EGFR mutation status in tissue and plasma for predicting response to EGFR-TKIs in advanced non-small-cell lung cancer. Oncol Lett. 2017;13:2425–31.CrossRefGoogle Scholar
  26. 26.
    Mack PC, Holland WS, Burich RA, Sangha R, Solis LJ, Li Y, et al. EGFR mutations detected in plasma are associated with patient outcomes in erlotinib plus docetaxel-treated non-small cell lung cancer. J Thorac Oncol. 2009;4:1466–72.CrossRefGoogle Scholar

Copyright information

© Association of Clinical Biochemists of India 2019

Authors and Affiliations

  1. 1.Stem Cell Biology Lab, Department of Cancer BiologyGujarat Cancer and Research InstituteAhmedabadIndia
  2. 2.Medicinal Chemistry and Pharmacogenomics Division, Department of Cancer BiologyGujarat Cancer and Research InstituteAhmedabadIndia
  3. 3.Medical Oncology DepartmentThe Gujarat Cancer & Research InstituteAhmedabadIndia

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