Journal of Gastroenterology

, Volume 54, Issue 2, pp 108–121 | Cite as

Tumor-specific genetic aberrations in cell-free DNA of gastroesophageal cancer patients

  • Kristina Magaard KoldbyEmail author
  • Michael Bau Mortensen
  • Sönke Detlefsen
  • Per Pfeiffer
  • Mads Thomassen
  • Torben A. Kruse


The applicability of liquid biopsies is studied intensively in all types of cancer and analysis of circulating tumor DNA (ctDNA) has recently been implemented clinically for mutation detection in lung cancer. ctDNA may provide information about tumor quantity and mutations present in the tumor, and as such have many potential applications in diagnosis and treatment of cancer. It has been suggested that ctDNA analysis may overcome the issue of intra-tumor heterogeneity faced by tissue biopsies and serve as an additional diagnostic tool. Furthermore, liquid biopsies are potentially helpful for monitoring of treatment response as well as detection of minimal residual disease and relapse. Gastroesophageal cancers (GEC) have high mortality rates and the majority of patients present with advanced stage at diagnosis or succumb due to disease recurrence even after radical resection of the primary tumor. Biomarkers that can help optimize treatment strategy are thus highly desirable. The present study is a review of published data on ctDNA in GEC patients. We identified 25 studies in which tumor-specific genetic aberrations were investigated in plasma or serum and discuss these in relation to the methods applied for ctDNA analysis. The methods used for ctDNA detection greatly influence the sensitivity of the analysis and, therefore, the potential clinical applications. We found that studies of ctDNA in GEC, although limited in number, are promising for several applications such as genetic profiling of tumors and monitoring of disease progression. However, more studies are needed to establish if and how this analysis can be clinically implemented.


Circulating tumor DNA Cell-free DNA Biomarker Gastric cancer Esophageal cancer 



This work was supported by funding from Odense University Hospital (grants from Free Research Funds, “Overlægerådets forskningsfond”, and “Frontlinjepuljen”), University of Southern Denmark, and the Frimodt Heineke Foundation.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.


  1. 1.
    Ferlay J, Soerjomataram I, Dikshit R, et al. Cancer incidence and mortality worldwide: sources, methods and major patterns in GLOBOCAN 2012. Int J Cancer. 2015;136:359–86.CrossRefGoogle Scholar
  2. 2.
    Cunningham D, Allum WH, Stenning SP, et al. Perioperative chemotherapy versus surgery alone for resectable gastroesophageal cancer. New Engl J Med. 2006;355:11–20.CrossRefGoogle Scholar
  3. 3.
    Ychou M, Boige V, Pignon JP, et al. Perioperative chemotherapy compared with surgery alone for resectable gastroesophageal adenocarcinoma: an FNCLCC and FFCD multicenter phase III trial. J Clin Oncol. 2011;29:1715–21.CrossRefGoogle Scholar
  4. 4.
    Shapiro J, van Lanschot JJB, Hulshof M, et al. Neoadjuvant chemoradiotherapy plus surgery versus surgery alone for oesophageal or junctional cancer (CROSS): long-term results of a randomised controlled trial. Lancet Oncol. 2015;16:1090–8.CrossRefGoogle Scholar
  5. 5.
    Lordick F, Mariette C, Haustermans K, et al. Oesophageal cancer: ESMO clinical practice guidelines for diagnosis, treatment and follow-up. Ann Oncol. 2016;27:v50–7.CrossRefGoogle Scholar
  6. 6.
    Smyth EC, Verheij M, Allum W, et al. Gastric cancer: ESMO clinical practice guidelines for diagnosis, treatment and follow-up. Ann Oncol. 2016;27:v38–49.CrossRefGoogle Scholar
  7. 7.
    Fuchs CS, Tomasek J, Yong CJ, et al. Ramucirumab monotherapy for previously treated advanced gastric or gastro-oesophageal junction adenocarcinoma (REGARD): an international, randomised, multicentre, placebo-controlled, phase 3 trial. Lancet. 2014;383:31–9.CrossRefGoogle Scholar
  8. 8.
    Bang YJ, Van Cutsem E, Feyereislova A, et al. Trastuzumab in combination with chemotherapy versus chemotherapy alone for treatment of HER2-positive advanced gastric or gastro-oesophageal junction cancer (ToGA): a phase 3, open-label, randomised controlled trial. Lancet. 2010;376:687–97.CrossRefGoogle Scholar
  9. 9.
    Maron SB, Catenacci DV. Novel targeted therapies for esophagogastric cancer. Surg Oncol Clin N Am. 2017;26:293–312.CrossRefGoogle Scholar
  10. 10.
    Zhou J, Ma X, Bi F, et al. Clinical significance of circulating tumor cells in gastric cancer patients. Oncotarget. 2017;8:25713–20.Google Scholar
  11. 11.
    Armstrong D, Wildman DE. Extracellular vesicles and the promise of continuous liquid biopsies. J Pathol Transl Med. 2018;52:1–8.CrossRefGoogle Scholar
  12. 12.
    Wan JCM, Massie C, Garcia-Corbacho J, et al. Liquid biopsies come of age: towards implementation of circulating tumour DNA. Nat Rev Cancer. 2017;17:223–38.CrossRefGoogle Scholar
  13. 13.
    Matsuzaki J, Suzuki H. Circulating microRNAs as potential biomarkers to detect transformation of barrett’s oesophagus to oesophageal adenocarcinoma. BMJ Open Gastroenterol. 2017;4:e000160.CrossRefGoogle Scholar
  14. 14.
    Crowley E, Di Nicolantonio F, Loupakis F, et al. Liquid biopsy: monitoring cancer-genetics in the blood. Nat Rev Clin Oncol. 2013;10:472–84.CrossRefGoogle Scholar
  15. 15.
    Siravegna G, Marsoni S, Siena S, et al. Integrating liquid biopsies into the management of cancer. Nat Rev Clin Oncol. 2017;14:531–48.CrossRefGoogle Scholar
  16. 16.
    FDA. P150044—Cobas EGFR MUTATION TEST V2. In: premarket approval. US Food & Drug Administration. 2016. Accessed 9 May 2018.
  17. 17.
    EMA. Iressa: public assessment report—product information. In European Medicines Agency. 2016; Accessed 9 May 2018.
  18. 18.
    EMA. Tagrisso: public assessment report—product information. In: European Medicines Agency. 2016. Accessed 9 May 2018.
  19. 19.
    Siewert JR, Ott K. Are squamous and adenocarcinomas of the esophagus the same disease? Semin Radiat Oncol. 2007;17:38–44.CrossRefGoogle Scholar
  20. 20.
    Torre LA, Bray F, Siegel RL, et al. Global cancer statistics, 2012. CA Cancer J Clin. 2015;65:87–108.CrossRefGoogle Scholar
  21. 21.
    Zheng T, Mayne ST, Holford TR, et al. The time trend and age—period—cohort effects on incidence of adenocarcinoma of the stomach in connecticut from 1955–1989. Cancer. 1993;72:330–40.CrossRefGoogle Scholar
  22. 22.
    Cancer Genome Atlas Research Network, U. Analysis Working Group: Asan, B.C. Cancer Agency, et al. Integrated genomic characterization of oesophageal carcinoma. Nature. 2017;541:169–75.CrossRefGoogle Scholar
  23. 23.
    Hayakawa Y, Sethi N, Sepulveda AR, et al. Oesophageal adenocarcinoma and gastric cancer: should we mind the gap? Nat Rev Cancer. 2016;16:305–18.CrossRefGoogle Scholar
  24. 24.
    Van Cutsem E, Sagaert X, Topal B, et al. Gastric cancer. Lancet. 2016;388:2654–64.CrossRefGoogle Scholar
  25. 25.
    Suzuki H, Mori H. World trends for H. pylori eradication therapy and gastric cancer prevention strategy by H. pylori test-and-treat. J Gastroenterol. 2018;53:354–61.CrossRefGoogle Scholar
  26. 26.
    Parkin DM. The global health burden of infection-associated cancers in the year 2002. Int J Cancer. 2006;118:3030–44.CrossRefGoogle Scholar
  27. 27.
    Lauren P. The two histological main types of gastric carcinoma: diffuse and so-called intestinal-type carcinoma: an attempt at a histo-clinical classification. Acta Pathol Microbiol Scand. 1965;64:31–49.CrossRefGoogle Scholar
  28. 28.
    Cancer Genome Atlas Research, N., Comprehensive molecular characterization of gastric adenocarcinoma. Nature. 2014;513:202–9.CrossRefGoogle Scholar
  29. 29.
    Sohn BH, Hwang JE, Jang HJ, et al. Clinical significance of four molecular subtypes of gastric cancer identified by the cancer genome atlas project. Clin Cancer Res. 2017. Scholar
  30. 30.
    Apicella M, Corso S, Giordano S. Targeted therapies for gastric cancer: failures and hopes from clinical trials. Oncotarget. 2017;8:57654–69.CrossRefGoogle Scholar
  31. 31.
    Gullo I, Grillo F, Molinaro L, et al. Minimum biopsy set for HER2 evaluation in gastric and gastro-esophageal junction cancer. Endosc Int Open. 2015;3:E165–70.CrossRefGoogle Scholar
  32. 32.
    Stroun M, Lyautey J, Lederrey C, et al. About the possible origin and mechanism of circulating DNA apoptosis and active DNA release. Clin Chim Acta. 2001;313:139–42.CrossRefGoogle Scholar
  33. 33.
    Jahr S, Hentze H, Englisch S, 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.Google Scholar
  34. 34.
    Lo YM, Zhang J, Leung TN, et al. Rapid clearance of fetal DNA from maternal plasma. Am J Hum Genet. 1999;64:218–24.CrossRefGoogle Scholar
  35. 35.
    Botezatu I, Serdyuk O, Potapova G, et al. Genetic analysis of DNA excreted in urine: a new approach for detecting specific genomic DNA sequences from cells dying in an organism. Clin Chem. 2000;46:1078–84.Google Scholar
  36. 36.
    Bryzgunova OE, Laktionov PP. Extracellular nucleic acids in urine: sources, structure, diagnostic potential. Acta Naturae. 2015;7:48–54.Google Scholar
  37. 37.
    Mandel P, Metais P. Les acides nucléiques du plasma sanguin chez l’homme [French]. C R Seances Soc Biol Fil. 1948;142:241–3.Google Scholar
  38. 38.
    Stroun M, Anker P, Maurice P, et al. Neoplastic characteristics of the DNA found in the plasma of cancer patients. Oncology. 1989;46:318–22.CrossRefGoogle Scholar
  39. 39.
    Olmedillas-Lopez S, Garcia-Arranz M, Garcia-Olmo D. Current and emerging applications of droplet digital PCR in oncology. Mol Diagn Ther. 2017;21:493–510.CrossRefGoogle Scholar
  40. 40.
    Hudecova I. Digital PCR analysis of circulating nucleic acids. Clin Biochem. 2015;48:948–56.CrossRefGoogle Scholar
  41. 41.
    Feng Q, Yang ZY, Zhang JT, et al. Comparison of direct sequencing and amplification refractory mutation system for detecting epidermal growth factor receptor mutation in non-small-cell lung cancer patients: a systematic review and meta-analysis. Oncotarget. 2017;8:59552–62.Google Scholar
  42. 42.
    Mauger F, How-Kit A, Tost J. COLD-PCR Technologies in the area of personalized medicine: methodology and applications. Mol Diagn Ther. 2017;21:269–83.CrossRefGoogle Scholar
  43. 43.
    Jovelet C, Ileana E, Le Deley MC, et al. Circulating cell-free tumor dna analysis of 50 genes by next-generation sequencing in the prospective MOSCATO trial. Clin Cancer Res. 2016;22:2960–8.CrossRefGoogle Scholar
  44. 44.
    Schwaederle M, Chattopadhyay R, Kato S, et al. Genomic alterations in circulating tumor DNA from diverse cancer patients identified by next-generation sequencing. Cancer Res. 2017;77:5419–27.CrossRefGoogle Scholar
  45. 45.
    Newman AM, Bratman SV, To J, et al. An ultrasensitive method for quantitating circulating tumor DNA with broad patient coverage. Nat Med. 2014;20:548–54.CrossRefGoogle Scholar
  46. 46.
    Kinde I, Wu J, Papadopoulos N, et al. Detection and quantification of rare mutations with massively parallel sequencing. Proc Natl Acad Sci USA. 2011;108:9530–5.CrossRefGoogle Scholar
  47. 47.
    Newman AM, Lovejoy AF, Klass DM, et al. Integrated digital error suppression for improved detection of circulating tumor DNA. Nat Biotechnol. 2016;34:547–55.CrossRefGoogle Scholar
  48. 48.
    Kukita Y, Matoba R, Uchida J, et al. High-fidelity target sequencing of individual molecules identified using barcode sequences: de novo detection and absolute quantitation of mutations in plasma cell-free DNA from cancer patients. DNA Res. 2015;22:269–77.CrossRefGoogle Scholar
  49. 49.
    Olsson E, Winter C, George A, et al. Serial monitoring of circulating tumor DNA in patients with primary breast cancer for detection of occult metastatic disease. EMBO Mol Med. 2015;7:1034–47.CrossRefGoogle Scholar
  50. 50.
    Reinert T, Scholer LV, Thomsen R, et al. Analysis of circulating tumour DNA to monitor disease burden following colorectal cancer surgery. Gut. 2015. Scholar
  51. 51.
    Heitzer E, Ulz P, Geigl JB, et al. Non-invasive detection of genome-wide somatic copy number alterations by liquid biopsies. Mol Oncol. 2016;10:494–502.CrossRefGoogle Scholar
  52. 52.
    Hovelson DH, Liu CJ, Wang Y, et al. Rapid, ultra low coverage copy number profiling of cell-free DNA as a precision oncology screening strategy. Oncotarget. 2017;8:89848–66.CrossRefGoogle Scholar
  53. 53.
    Milbury CA, Zhong Q, Lin J, et al. Determining lower limits of detection of digital PCR assays for cancer-related gene mutations. Biomol Detect Quantif. 2014;1:8–22.CrossRefGoogle Scholar
  54. 54.
    Ulz P, Heitzer E, Geigl JB, et al. Patient monitoring through liquid biopsies using circulating tumor DNA. Int J Cancer. 2017;141:887–96.CrossRefGoogle Scholar
  55. 55.
    Volckmar AL, Sultmann H, Riediger A, et al. A field guide for cancer diagnostics using cell-free DNA: from principles to practice and clinical applications. Genes Chromosomes Cancer. 2018;57:123–39.CrossRefGoogle Scholar
  56. 56.
    Bettegowda C, Sausen M, Leary RJ, et al. Detection of circulating tumor DNA in early- and late-stage human malignancies. Sci Transl Med. 2014;6:224ra24.CrossRefGoogle Scholar
  57. 57.
    Luo H, Li H, Hu Z, et al. Noninvasive diagnosis and monitoring of mutations by deep sequencing of circulating tumor DNA in esophageal squamous cell carcinoma. Biochem Biophys Res Commun. 2016;471:596–602.CrossRefGoogle Scholar
  58. 58.
    Ueda M, Iguchi T, Masuda T, et al. Somatic mutations in plasma cell-free DNA are diagnostic markers for esophageal squamous cell carcinoma recurrence. Oncotarget. 2016;7:62280–91.CrossRefGoogle Scholar
  59. 59.
    Hamakawa T, Kukita Y, Kurokawa Y, et al. Monitoring gastric cancer progression with circulating tumour DNA. Br J Cancer. 2015;112:352–6.CrossRefGoogle Scholar
  60. 60.
    Fang WL, Lan YT, Huang KH, et al. Clinical significance of circulating plasma DNA in gastric cancer. Int J Cancer. 2016;138:2974–83.CrossRefGoogle Scholar
  61. 61.
    Gao J, Wang H, Zang W, et al. Circulating tumor DNA functions as an alternative for tissue to overcome tumor heterogeneity in advanced gastric cancer. Cancer Sci. 2017;108(9):1881–7.CrossRefGoogle Scholar
  62. 62.
    Song Y, Li L, Ou Y, et al. Identification of genomic alterations in oesophageal squamous cell cancer. Nature. 2014;509:91–5.CrossRefGoogle Scholar
  63. 63.
    Gao YB, Chen ZL, Li JG, et al. Genetic landscape of esophageal squamous cell carcinoma. Nat Genet. 2014;46:1097–102.CrossRefGoogle Scholar
  64. 64.
    Wang K, Johnson A, Ali SM, et al. Comprehensive genomic profiling of advanced esophageal squamous cell carcinomas and esophageal adenocarcinomas reveals similarities and differences. Oncologist. 2015;20:1132–9.CrossRefGoogle Scholar
  65. 65.
    Takeshita H, Ichikawa D, Komatsu S, et al. Prediction of CCND1 amplification using plasma DNA as a prognostic marker in oesophageal squamous cell carcinoma. Br J Cancer. 2010;102:1378–83.CrossRefGoogle Scholar
  66. 66.
    Komatsu S, Ichikawa D, Hirajima S, et al. Clinical impact of predicting CCND1 amplification using plasma DNA in superficial esophageal squamous cell carcinoma. Dig Dis Sci. 2014;59:1152–9.CrossRefGoogle Scholar
  67. 67.
    Press MF, Ellis CE, Gagnon RC, et al. HER2 Status in advanced or metastatic gastric, esophageal, or gastroesophageal adenocarcinoma for entry to the TRIO-013/LOGiC Trial of Lapatinib. Mol Cancer Ther. 2017;16:228–38.CrossRefGoogle Scholar
  68. 68.
    Andolfo I, Petrosino G, Vecchione L, et al. Detection of erbB2 copy number variations in plasma of patients with esophageal carcinoma. BMC Cancer. 2011;11:126.CrossRefGoogle Scholar
  69. 69.
    Park KU, Lee HE, Nam SK, et al. The quantification of HER2 and MYC gene fragments in cell-free plasma as putative biomarkers for gastric cancer diagnosis. Clin Chem Lab Med. 2014;52:1033–40.Google Scholar
  70. 70.
    Kinugasa H, Nouso K, Tanaka T, et al. Droplet digital PCR measurement of HER2 in patients with gastric cancer. Br J Cancer. 2015;112:1652–5.CrossRefGoogle Scholar
  71. 71.
    Lee HE, Park KU, Yoo SB, et al. Clinical significance of intratumoral HER2 heterogeneity in gastric cancer. Eur J Cancer. 2013;49:1448–57.CrossRefGoogle Scholar
  72. 72.
    Shoda K, Ichikawa D, Fujita Y, et al. Monitoring the HER2 copy number status in circulating tumor DNA by droplet digital PCR in patients with gastric cancer. Gastric Cancer. 2017;20:126–35.CrossRefGoogle Scholar
  73. 73.
    El Messaoudi S, Rolet F, Mouliere F, et al. Circulating cell free DNA: preanalytical considerations. Clin Chim Acta. 2013;424:222–30.CrossRefGoogle Scholar
  74. 74.
    Shoda K, Masuda K, Ichikawa D, et al. HER2 amplification detected in the circulating DNA of patients with gastric cancer: a retrospective pilot study. Gastric Cancer. 2015;18:698–710.CrossRefGoogle Scholar
  75. 75.
    Park KU, Lee HE, Park DJ, et al. MYC quantitation in cell-free plasma DNA by real-time PCR for gastric cancer diagnosis. Clin Chem Lab Med. 2009;47:530–6.Google Scholar
  76. 76.
    Pectasides E, Stachler MD, Derks S, et al. Genomic heterogeneity as a barrier to precision medicine in gastroesophageal adenocarcinoma. Cancer Discov. 2018;8:37–48.CrossRefGoogle Scholar
  77. 77.
    Bang Y-J, Cutsem EV, Mansoor W, et al. A randomized, open-label phase II study of AZD4547 (AZD) versus paclitaxel (P) in previously treated patients with advanced gastric cancer (AGC) with fibroblast growth factor receptor 2 (FGFR2) polysomy or gene amplification (amp): SHINE study. J Clin Oncol. 2015;33:4014.CrossRefGoogle Scholar
  78. 78.
    Smyth E, Turner NC, Pearson A, et al. Phase II study of AZD4547 in FGFR amplified tumours: Gastroesophageal cancer (GC) cohort pharmacodynamic and biomarker results. J Clin Oncol. 2016. Scholar
  79. 79.
    Pearson A, Smyth E, Babina IS, et al. High-Level clonal FGFR amplification and response to FGFR inhibition in a translational clinical trial. Cancer Discov. 2016;6:838–51.CrossRefGoogle Scholar
  80. 80.
    Rumiato E, Boldrin E, Malacrida S, et al. Detection of genetic alterations in cfDNA as a possible strategy to monitor the neoplastic progression of Barrett’s esophagus. Transl Res. 2017;190(16–24):e1.Google Scholar
  81. 81.
    Eisenberger CF, Knoefel WT, Peiper M, et al. Squamous cell carcinoma of the esophagus can be detected by microsatellite analysis in tumor and serum. Clin Cancer Res. 2003;9:4178–83.Google Scholar
  82. 82.
    Eisenberger CF, Stoecklein NH, Jazra S, et al. The detection of oesophageal adenocarcinoma by serum microsatellite analysis. Eur J Surg Oncol. 2006;32:954–60.CrossRefGoogle Scholar
  83. 83.
    Ma X, Zhu L, Wu X, et al. Cell-Free DNA provides a good representation of the tumor genome despite its biased fragmentation patterns. PLoS One. 2017;12:e0169231.CrossRefGoogle Scholar
  84. 84.
    Snyder MW, Kircher M, Hill AJ, et al. Cell-free DNA comprises an in vivo nucleosome footprint that informs its tissues-of-origin. Cell. 2016;164:57–68.CrossRefGoogle Scholar
  85. 85.
    Diehl F, Schmidt K, Choti MA, et al. Circulating mutant DNA to assess tumor dynamics. Nat Med. 2008;14:985–90.CrossRefGoogle Scholar
  86. 86.
    Shi XQ, Xue WH, Zhao SF, et al. Dynamic tracing for epidermal growth factor receptor mutations in urinary circulating DNA in gastric cancer patients. Tumour Biol. 2017;39:1–5.Google Scholar
  87. 87.
    Kwak EL, Ahronian LG, Siravegna G, et al. Molecular heterogeneity and receptor coamplification drive resistance to targeted therapy in MET-Amplified esophagogastric cancer. Cancer Discov. 2015;5:1271–81.CrossRefGoogle Scholar
  88. 88.
    Petty RD, Dahle-Smith A, Stevenson DAJ, et al. Gefitinib and EGFR gene copy number aberrations in esophageal cancer. J Clin Oncol. 2017;35:2279–87.CrossRefGoogle Scholar
  89. 89.
    Du J, Wu X, Tong X, et al. Circulating tumor DNA profiling by next generation sequencing reveals heterogeneity of crizotinib resistance mechanisms in a gastric cancer patient with MET amplification. Oncotarget. 2017;8:26281–7.Google Scholar
  90. 90.
    Garcia-Murillas I, Schiavon G, Weigelt B, et al. Mutation tracking in circulating tumor DNA predicts relapse in early breast cancer. Sci Transl Med. 2015;7:302ra133.CrossRefGoogle Scholar
  91. 91.
    Aurello P, Petrucciani N, Antolino L, et al. Follow-up after curative resection for gastric cancer: is it time to tailor it? World J Gastroenterol. 2017;23:3379–87.CrossRefGoogle Scholar
  92. 92.
    Takahashi Y, Takeuchi T, Sakamoto J, et al. The usefulness of CEA and/or CA19-9 in monitoring for recurrence in gastric cancer patients: a prospective clinical study. Gastric Cancer. 2003;6:142–5.CrossRefGoogle Scholar

Copyright information

© Japanese Society of Gastroenterology 2018

Authors and Affiliations

  1. 1.Department of Clinical GeneticsOdense University HospitalOdenseDenmark
  2. 2.Human Genetics, Department of Clinical ResearchUniversity of Southern DenmarkOdenseDenmark
  3. 3.Department of SurgeryOdense University HospitalOdenseDenmark
  4. 4.Department of PathologyOdense University HospitalOdenseDenmark
  5. 5.Department of OncologyOdense University HospitalOdenseDenmark

Personalised recommendations