A synonymous EGFR polymorphism predicting responsiveness to anti-EGFR therapy in metastatic colorectal cancer patients

Abstract

Genetic factors are known to affect the efficiency of therapy with monoclonal antibodies (mAbs) targeting the epidermal growth factor receptor (EGFR) in patients with metastatic colorectal cancer (mCRC). At present, the only accepted molecular marker predictive of the response to anti-EGFR mAbs is the somatic mutation of KRAS and NRAS as a marker of resistance to anti-EGFR. However, only a fraction of KRAS wild-type patients benefit from that treatment. In this study, we show that the EGFR gene polymorphism rs1050171 defines, independently of RAS mutational status, a sub-population of 11 % of patients with a better clinical outcome after anti-EGFR treatment. Median PFS for patients with the GG genotype was 10.17 months compared to 5.37 of those with AG + AA genotypes. Taken together, our findings could be used to better define CRC populations responding to anti-EGFR therapy. Further studies in larger independent cohorts are necessary to validate the present observation that a synonymous polymorphism in EGFR gene impacts on clinical responsiveness.

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References

  1. 1.

    Jonker DJ, O’Callaghan CJ, Karapetis CS, Zalcberg JR, Tu D, Au HJ, et al. Cetuximab for the treatment of colorectal cancer. N Engl J Med. 2007;357(20):2040–8. doi:10.1056/NEJMoa071834.

    CAS  Article  PubMed  Google Scholar 

  2. 2.

    Van Cutsem E, Kohne CH, Hitre E, Zaluski J, Chang Chien CR, Makhson A, et al. Cetuximab and chemotherapy as initial treatment for metastatic colorectal cancer. N Engl J Med. 2009;360(14):1408–17.

    Article  PubMed  Google Scholar 

  3. 3.

    Di Fiore F, Sesboue R, Michel P, Sabourin JC, Frebourg T. Molecular determinants of anti-EGFR sensitivity and resistance in metastatic colorectal cancer. Br J Cancer. 2010;103(12):1765–72. doi:10.1038/sj.bjc.6606008.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. 4.

    Bardelli A, Siena S. Molecular mechanisms of resistance to cetuximab and panitumumab in colorectal cancer. J Clin Oncol. 2010;28(7):1254–61.

    CAS  Article  PubMed  Google Scholar 

  5. 5.

    Meriggi F, Vermi W, Bertocchi P, Zaniboni A. The emerging role of NRAS mutations in colorectal cancer patients selected for anti-EGFR therapies. Rev Recent Clin Trials. 2014.

  6. 6.

    Jacobs B, De Roock W, Piessevaux H, Van Oirbeek R, Biesmans B, De Schutter J, et al. Amphiregulin and epiregulin mRNA expression in primary tumors predicts outcome in metastatic colorectal cancer treated with cetuximab. J Clin Oncol. 2009;27(30):5068–74. doi:10.1200/JCO.2008.21.3744.

    CAS  Article  PubMed  Google Scholar 

  7. 7.

    Leto SM, Trusolino L. Primary and acquired resistance to EGFR-targeted therapies in colorectal cancer: impact on future treatment strategies. J Mol Med. 2014. doi:10.1007/s00109-014-1161-2.

    PubMed  PubMed Central  Google Scholar 

  8. 8.

    Therkildsen C, Bergmann TK, Henrichsen-Schnack T, Ladelund S, Nilbert M. The predictive value of KRAS, NRAS, BRAF, PIK3CA and PTEN for anti-EGFR treatment in metastatic colorectal cancer: a systematic review and meta-analysis. Acta Oncol. 2014. doi:10.3109/0284186X.2014.895036.

    Google Scholar 

  9. 9.

    Lai CY, Sung FC, Hsieh LL, Tang R, Chiou HY, Wu FY, et al. Associations between genetic polymorphisms of epidermal growth factor receptor (EGFR) and survival of colorectal cancer (CRC) patients treated with 5-fluorouracil-based chemotherapy. Ann Surg Oncol. 2013;20 Suppl 3:S599–606. doi:10.1245/s10434-013-3069-4.

    Article  PubMed  Google Scholar 

  10. 10.

    Zhang W, Stoehlmacher J, Park DJ, Yang D, Borchard E, Gil J, et al. Gene polymorphisms of epidermal growth factor receptor and its downstream effector, interleukin-8, predict oxaliplatin efficacy in patients with advanced colorectal cancer. Clin Colorectal Cancer. 2005;5(2):124–31.

    Article  PubMed  Google Scholar 

  11. 11.

    Garm Spindler KL, Pallisgaard N, Rasmussen AA, Lindebjerg J, Andersen RF, Cruger D, et al. The importance of KRAS mutations and EGF61A>G polymorphism to the effect of cetuximab and irinotecan in metastatic colorectal cancer. Ann Oncol. 2009;20(5):879–84. doi:10.1093/annonc/mdn712.

    Article  PubMed  Google Scholar 

  12. 12.

    Graziano F, Ruzzo A, Loupakis F, Canestrari E, Santini D, Catalano V, et al. Pharmacogenetic profiling for cetuximab plus irinotecan therapy in patients with refractory advanced colorectal cancer. J Clin Oncol. 2008;26(9):1427–34. doi:10.1200/JCO.2007.12.4602.

    CAS  Article  PubMed  Google Scholar 

  13. 13.

    Metzger B, Chambeau L, Begon DY, Faber C, Kayser J, Berchem G, et al. The human epidermal growth factor receptor (EGFR) gene in European patients with advanced colorectal cancer harbors infrequent mutations in its tyrosine kinase domain. BMC Med Genet. 2011;12:144. doi:10.1186/1471-2350-12-144.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  14. 14.

    Basili G, Bonin S, Dotti I. DNA sequencing from PCR products. In: Stanta G, editor. Guidelines for molecular analysis in archive tissues. Berlin: Springer; 2011. p. 135–41.

    Google Scholar 

  15. 15.

    Hlubek F, Jung A. Fast protocol for DNA extraction from formalin-fixed paraffin-embedded tissues. In: Stanta G, editor. Guidelines for molecular analyses in archive tissues. Berlin: Springer; 2011. p. 41–3.

    Google Scholar 

  16. 16.

    Bonin S, Stanta G. RNA extraction from formalin-fixed paraffin-embedded tissues. In: Stanta G, editor. Guidelines for molecular analysis in archive tissues. 1st ed. Berlin: Springer-Verlag; 2011. p. 57–61.

    Google Scholar 

  17. 17.

    Nardon E, Donada M, Bonin S, Dotti I, Stanta G. Higher random oligo concentration improves reverse transcription yield of cDNA from bioptic tissues and quantitative RT-PCR reliability. Exp Mol Pathol. 2009;87(2):146–51. doi:10.1016/j.yexmp.2009.07.005.

    CAS  Article  PubMed  Google Scholar 

  18. 18.

    Donada M, Bonin S, Nardon E, De Pellegrin A, Decorti G, Stanta G. Thymidilate synthase expression predicts longer survival in patients with stage II colon cancer treated with 5-flurouracil independently of microsatellite instability. J Cancer Res Clin Oncol. 2011;137(2):201–10.

    CAS  Article  PubMed  Google Scholar 

  19. 19.

    EGAPP WG. Recommendations from the EGAPP Working Group: can testing of tumor tissue for mutations in EGFR pathway downstream effector genes in patients with metastatic colorectal cancer improve health outcomes by guiding decisions regarding anti-EGFR therapy? Genet Med. 2013;15(7):517–27. doi:10.1038/gim.2012.184.

    Article  CAS  Google Scholar 

  20. 20.

    Gil Ferreira C, Aran V, Zalcberg-Renault I, Victorino AP, Salem JH, Bonamino MH, et al. KRAS mutations: variable incidences in a Brazilian cohort of 8,234 metastatic colorectal cancer patients. BMC Gastroenterol. 2014;14:73. doi:10.1186/1471-230X-14-73.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. 21.

    De Roock W, Jonker DJ, Di Nicolantonio F, Sartore-Bianchi A, Tu D, Siena S, et al. Association of KRAS p.G13D mutation with outcome in patients with chemotherapy-refractory metastatic colorectal cancer treated with cetuximab. JAMA. 2010;304(16):1812–20. doi:10.1001/jama.2010.1535.

    Article  PubMed  Google Scholar 

  22. 22.

    Choi JE, Park SH, Kim KM, Lee WK, Kam S, Cha SI, et al. Polymorphisms in the epidermal growth factor receptor gene and the risk of primary lung cancer: a case-control study. BMC Cancer. 2007;7:199.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. 23.

    Wang Y, Bao W, Shi H, Jiang C, Zhang Y. Epidermal growth factor receptor exon 20 mutation increased in post-chemotherapy patients with non-small cell lung cancer detected with patients’ blood samples. Transl Oncol. 2013;6(4):504–10.

    Article  PubMed  PubMed Central  Google Scholar 

  24. 24.

    Zhang W, Stabile LP, Keohavong P, Romkes M, Grandis JR, Traynor AM, et al. Mutation and polymorphism in the EGFR-TK domain associated with lung cancer. J Thorac Oncol. 2006;1(7):635–47.

    PubMed  Google Scholar 

  25. 25.

    Sasaki H, Endo K, Takada M, Kawahara M, Tanaka H, Kitahara N, et al. EGFR polymorphism of the kinase domain in Japanese lung cancer. J Surg Res. 2008;148(2):260–3.

    CAS  Article  PubMed  Google Scholar 

  26. 26.

    Marx AH, Zielinski M, Kowitz CM, Dancau AM, Thieltges S, Simon R et al. Homogeneous EGFR amplification defines a subset of aggressive Barrett’s adenocarcinomas with poor prognosis. Histopathology. 2010.

  27. 27.

    Kaneko K, Kumekawa Y, Makino R, Nozawa H, Hirayama Y, Kogo M, et al. EGFR gene alterations as a prognostic biomarker in advanced esophageal squamous cell carcinoma. Front Biosci. 2010;15:65–72.

    CAS  Article  Google Scholar 

  28. 28.

    Weiss GA, Rossi MR, Khushalani NI, Lo K, Gibbs JF, Bharthuar A, et al. Evaluation of phosphatidylinositol-3-kinase catalytic subunit (PIK3CA) and epidermal growth factor receptor (EGFR) gene mutations in pancreaticobiliary adenocarcinoma. J Gastrointest Oncol. 2013;4(1):20–9. doi:10.3978/j.issn.2078-6891.2012.012.

    CAS  PubMed  PubMed Central  Google Scholar 

  29. 29.

    Zhang J, Zhan Z, Wu J, Zhang C, Yang Y, Tong S, et al. Association among polymorphisms in EGFR gene exons, lifestyle and risk of gastric cancer with gender differences in Chinese Han subjects. PLoS One. 2013;8(3):e59254. doi:10.1371/journal.pone.0059254.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  30. 30.

    Parmley JL, Hurst LD. How do synonymous mutations affect fitness? BioEssays. 2007;29(6):515–9. doi:10.1002/bies.20592.

    CAS  Article  PubMed  Google Scholar 

  31. 31.

    Nakazaki K, Kato Y, Taguchi T, Inayama Y, Ishiguro Y, Kondo N, et al. Heterozygous mutation (G/G→G/A) at nt 2607 of the EGFR gene is closely associated with increases in EGFR copy number and mRNA half life, but impaired EGFR protein synthesis in squamous cell carcinomas of the head and neck—implication for gefitinib efficacy. Oncol Lett. 2010;1(6):1017–20. doi:10.3892/ol.2010.188.

    CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgments

The authors would like to thank Dr Valentina Melita for the English revision of the manuscript.

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Correspondence to Giorgio Stanta.

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This study was partially supported by the FRA2011 grant from the University of Trieste.

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Bonin, S., Donada, M., Bussolati, G. et al. A synonymous EGFR polymorphism predicting responsiveness to anti-EGFR therapy in metastatic colorectal cancer patients. Tumor Biol. 37, 7295–7303 (2016). https://doi.org/10.1007/s13277-015-4543-3

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Keywords

  • mCRC
  • RAS mutation
  • EGFR gene polymorphism rs1050171
  • Synonymous SNP
  • FFPE
  • Survival