Gastroesophageal adenocarcinoma (GOA) has poor clinical outcomes and lacks reliable blood markers. Here we present circulating tumour DNA (ctDNA) as an emerging biomarker.
Forty patients (17 palliative and 23 curative) were followed by serial plasma monitoring. Primary tumour DNA was analysed by targeted next-generation sequencing to identify somatic single-nucleotide variants (SNVs), and Nanostring nCounter® to detect copy number alterations (CNAs). Patient-specific SNVs and CNA amplifications (CNAamp) were analysed in plasma using digital droplet PCR and quantitative PCR, respectively.
Thirty-five patients (13 palliative, 22 curative) had ≥1 SNVs and/or CNAamp detected in primary tumour DNA suitable for tracking in plasma. Eighteen of 35 patients (nine palliative, nine curative) had ≥1 ctDNA-positive plasma sample. Detection of postoperative ctDNA predicted short RFS (190 vs 934 days, HR = 3.7, p = 0.028) and subsequent relapse (PPV for relapse 0.83). High ctDNA levels (>60.5 copies/ml) at diagnosis of metastatic disease predicted poor OS (90 vs 372 days, HR = 11.7 p < 0.001).
Sensitive ctDNA detection allows disease monitoring and prediction of short OS in metastatic patients. Presence of ctDNA postoperatively predicts relapse and defines a ‘molecular relapse’ before overt clinical disease. This lead time defines a potential therapeutic window for additional anticancer therapy.
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Bray, F., Ferlay, J., Soerjomataram, I., Siegel, R. L., Torre, L. A. & Jemal, A. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J. Clin. 68, 394–424 (2018).
Thrift, A. P. The epidemic of oesophageal carcinoma: where are we now? Cancer Epidemiol. 41, 88–95 (2016).
Hartgrink, H. H., Jansen, E. P., van Grieken, N. C. & van de Velde, C. J. Gastric cancer. Lancet 374, 477–490 (2009).
Cancer Genome Atlas Research N, Analysis Working Group, Asan, U., BCC, Agency, Brigham, Women’s, H., Broad, I. et al. Integrated genomic characterization of oesophageal carcinoma. Nature 541, 169–175 (2017).
Cunningham, D., Starling, N., Rao, S., Iveson, T., Nicolson, M., Coxon, F. et al. Capecitabine and oxaliplatin for advanced esophagogastric cancer. N. Engl. J. Med. 358, 36–46 (2008).
Bang, Y. J., Van Cutsem, E., Feyereislova, A., Chung, H. C., Shen, L., Sawaki, 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 376, 687–697 (2010).
van Hagen, P., Hulshof, M. C., van Lanschot, J. J., Steyerberg, E. W., van Berge Henegouwen, M. I., Wijnhoven, B. P. et al. Preoperative chemoradiotherapy for esophageal or junctional cancer. N. Engl. J. Med. 366, 2074–2084 (2012).
Cunningham, D., Allum, W. H., Stenning, S. P., Thompson, J. N., Van de Velde, C. J., Nicolson, M. et al. Perioperative chemotherapy versus surgery alone for resectable gastroesophageal cancer. N. Engl. J. Med. 355, 11–20 (2006).
Al-Batran, S.-E., Homann, N., Pauligk, C., Goetze, T. O., Meiler, J., Kasper, S. et al. Perioperative chemotherapy with fluorouracil plus leucovorin, oxaliplatin, and docetaxel versus fluorouracil or capecitabine plus cisplatin and epirubicin for locally advanced, resectable gastric or gastro-oesophageal junction adenocarcinoma (FLOT4): a randomised, phase 2/3 trial. Lancet 393, 1948–1957 (2019).
Ychou, M., Boige, V., Pignon, J. P., Conroy, T., Bouche, O., Lebreton, G. et al. Perioperative chemotherapy compared with surgery alone for resectable gastroesophageal adenocarcinoma: an FNCLCC and FFCD multicenter phase III trial. J. Clin. Oncol. 29, 1715–1721 (2011).
Fribbens, C., Garcia Murillas, I., Beaney, M., Hrebien, S., O’Leary, B., Kilburn, L. et al. Tracking evolution of aromatase inhibitor resistance with circulating tumour DNA analysis in metastatic breast cancer. Ann. Oncol. 29, 145–153 (2018).
Diehl, F., Schmidt, K., Choti, M. A., Romans, K., Goodman, S., Li, M. et al. Circulating mutant DNA to assess tumor dynamics. Nat. Med. 14, 985–990 (2008).
Ni, J., Weng, L., Liu, Y., Sun, Z., Bai, C. & Wang, Y. Dynamic monitoring of EGFR mutations in circulating cell-free DNA for EGFR-mutant metastatic patients with lung cancer: early detection of drug resistance and prognostic significance. Oncol. Lett. 13, 4549–4557 (2017).
Dawson, S. J., Tsui, D. W., Murtaza, M., Biggs, H., Rueda, O. M., Chin, S. F. et al. Analysis of circulating tumor DNA to monitor metastatic breast cancer. N. Engl. J. Med. 368, 1199–1209 (2013).
Garcia-Murillas, I., Schiavon, G., Weigelt, B., Ng, C., Hrebien, S., Cutts, R. J. et al. Mutation tracking in circulating tumor DNA predicts relapse in early breast cancer. Sci. Transl. Med. 7, 302ra133 (2015).
Abbosh, C., Birkbak, N. J., Wilson, G. A., Jamal-Hanjani, M., Constantin, T., Salari, R. et al. Phylogenetic ctDNA analysis depicts early-stage lung cancer evolution. Nature 545, 446–451 (2017).
Pectasides, E., Stachler, M. D., Derks, S., Liu, Y., Maron, S., Islam, M. et al. Genomic heterogeneity as a barrier to precision medicine in gastroesophageal adenocarcinoma. Cancer Discov. 8, 37–48 (2018).
Gao, J., Wang, H., Zang, W., Li, B., Rao, G., Li, L. et al. Circulating tumor DNA functions as an alternative for tissue to overcome tumor heterogeneity in advanced gastric cancer. Cancer Sci. 108, 1881–1887 (2017).
Bedard, P. L., Hansen, A. R., Ratain, M. J. & Siu, L. L. Tumour heterogeneity in the clinic. Nature 501, 355 (2013).
Dulak, A. M., Stojanov, P., Peng, S., Lawrence, M. S., Fox, C., Stewart, C. et al. Exome and whole-genome sequencing of esophageal adenocarcinoma identifies recurrent driver events and mutational complexity. Nat. Genet. 45, 478–486 (2013).
Murugaesu, N., Wilson, G. A., Birkbak, N. J., Watkins, T. B., McGranahan, N., Kumar, S. et al. Tracking the genomic evolution of esophageal adenocarcinoma through neoadjuvant chemotherapy. Cancer Discov. 5, 821–831 (2015).
Kato, S., Okamura, R., Baumgartner, J. M., Patel, H., Leichman, L., Kelly, K. et al. Analysis of circulating tumor DNA and clinical correlates in patients with esophageal, gastroesophageal junction, and gastric adenocarcinoma. Clin. Cancer Res. 24, 6248–6256 (2018).
Pasternack, H., Fassunke, J., Plum, P. S., Chon, S. H., Hescheler, D. A., Gassa, A. et al. Somatic alterations in circulating cell-free DNA of oesophageal carcinoma patients during primary staging are indicative for post-surgical tumour recurrence. Sci. Rep. 8, 14941 (2018).
Hamakawa, T., Kukita, Y., Kurokawa, Y., Miyazaki, Y., Takahashi, T., Yamasaki, M. et al. Monitoring gastric cancer progression with circulating tumour DNA. Br. J. Cancer 112, 352–356 (2014).
Shoda, K., Ichikawa, D., Fujita, Y., Masuda, K., Hiramoto, H., Hamada, J. et al. Monitoring the HER2 copy number status in circulating tumor DNA by droplet digital PCR in patients with gastric cancer. Gastric Cancer 20, 126–135 (2017).
Maron, S. B., Chase, L. M., Lomnicki, S., Kochanny, S., Moore, K. L., Joshi, S. S. et al. Circulating tumor DNA sequencing analysis of gastroesophageal adenocarcinoma. Clin. Cancer Res. 25, 7098–7112 (2019).
Davidson, M., Barber, L. J., Woolston, A., Cafferkey, C., Mansukhani, S., Griffiths, B. et al. Detecting and tracking circulating tumour DNA copy number profiles during first line chemotherapy in oesophagogastric adenocarcinoma. Cancers (Basel) 11, 1-18. https://doi.org/10.3390/cancers11050736 (2019).
Page, K., Powles, T., Slade, M. J., DEB, M. T., Walker, R. A., Coombes, R. C. et al. The importance of careful blood processing in isolation of cell-free DNA. Ann. N. Y. Acad. Sci. 1075, 313–317 (2006).
Page, K., Guttery, D. S., Zahra, N., Primrose, L., Elshaw, S. R., Pringle, J. H. et al. Influence of plasma processing on recovery and analysis of circulating nucleic acids. PLoS ONE 8, e77963 (2013).
Cancer Genome Atlas Research N. Comprehensive molecular characterization of gastric adenocarcinoma. Nature 513, 202–209 (2014).
Gao, J., Aksoy, B. A., Dogrusoz, U., Dresdner, G., Gross, B., Sumer, S. O. et al. Integrative analysis of complex cancer genomics and clinical profiles using the cBioPortal. Sci. Signal 6, pl1 (2013).
Cerami, E., Gao, J., Dogrusoz, U., Gross, B. E., Sumer, S. O., Aksoy, B. A. et al. The cBio cancer genomics portal: an open platform for exploring multidimensional cancer genomics data. Cancer Discov. 2, 401–404 (2012).
Scientific, T. F. Ion ampliseq designer. https://ampliseq.com (2015).
Guttery, D. S., Page, K., Hills, A., Woodley, L., Marchese, S. D., Rghebi, B. et al. Noninvasive detection of activating estrogen receptor 1 (ESR1) mutations in estrogen receptor-positive metastatic breast cancer. Clin. Chem. 61, 974–982 (2015).
Page, K., Hava, N., Ward, B., Brown, J., Guttery, D. S., Ruangpratheep, C. et al. Detection of HER2 amplification in circulating free DNA in patients with breast cancer. Br. J. Cancer 104, 1342–1348 (2011).
Geiss, G. K., Bumgarner, R. E., Birditt, B., Dahl, T., Dowidar, N., Dunaway, D. L. et al. Direct multiplexed measurement of gene expression with color-coded probe pairs. Nat. Biotechnol. 26, 317–325 (2008).
Shaw, J. A., Guttery, D. S., Hills, A., Fernandez-Garcia, D., Page, K., Rosales, B. M. et al. Mutation analysis of cell-free DNA and single circulating tumor cells in metastatic breast cancer patients with high CTC counts. Clin. Cancer Res. 23, 88–96 (2017).
Jamal-Hanjani, M., Wilson, G. A., McGranahan, N., Birkbak, N. J., Watkins, T. B. K., Veeriah, S. et al. Tracking the evolution of non-small-cell lung cancer. N. Engl. J. Med. 376, 2109–2121 (2017).
Andersen, P. K. & Gill, R. D. Cox’s regression model for counting processes: a large sample study. Ann. Stat. 10, 1100–1120 (1982).
Hudecova, I. Digital PCR analysis of circulating nucleic acids. Clin. Biochem. 48, 948–956 (2015).
Couto, S. S. The pathologist’s slide reveals more than meets the eye: loss of heterozygosity and cancer biology. Vet. Pathol. 48, 236–244 (2011).
Butler, T. M., Johnson-Camacho, K., Peto, M., Wang, N. J., Macey, T. A., Korkola, J. E. et al. Exome sequencing of cell-free DNA from metastatic cancer patients identifies clinically actionable mutations distinct from primary disease. PLoS ONE 10, e0136407 (2015).
Toledo, R. A., Garralda, E., Mitsi, M., Pons, T., Monsech, J., Vega, E. et al. Exome sequencing of plasma DNA portrays the mutation landscape of ccolorectal cancer and discovers mutated VEGFR2 receptors as modulators of antiangiogenic therapies. Clin. Cancer Res. 24, 3550–3559 (2018).
Wang, Y., Zhao, C., Chang, L., Jia, R., Liu, R., Zhang, Y. et al. Circulating tumor DNA analyses predict progressive disease and indicate trastuzumab-resistant mechanism in advanced gastric cancer. EBioMedicine 43, 261–269 (2019).
Iqbal, M., Roberts, A., Starr, J., Mody, K. & Kasi, P. M. Feasibility and clinical value of circulating tumor DNA testing in patients with gastric adenocarcinomas. J. Gastrointest. Oncol. 10, 400–406 (2019).
Normando, S. R. C., Delgado, P. O., Rodrigues, A., David Filho, W. J., Fonseca, F. L. A., Cruz, F. et al. Circulating free plasma tumor DNA in patients with advanced gastric cancer receiving systemic chemotherapy. BMC Clin. Pathol. 18, 12 (2018).
Kim, J. Y., Park, D., Son, D. S., Nam, S. J., Kim, S. W., Jung, H. H. et al. Circulating tumor DNA shows variable clonal response of breast cancer during neoadjuvant chemotherapy. Oncotarget 8, 86423–86434 (2017).
Riva, F., Bidard, F. C., Houy, A., Saliou, A., Madic, J., Rampanou, A. et al. Patient-specific circulating tumor DNA detection during neoadjuvant chemotherapy in triple-negative breast cancer. Clin. Chem. 63, 691–699 (2017).
Tie, J., Cohen, J. D., Wang, Y., Li, L., Christie, M., Simons, K. et al. Serial circulating tumour DNA analysis during multimodality treatment of locally advanced rectal cancer: a prospective biomarker study. Gut 68, 663–671 (2019).
Clinicaltrials.gov. Prediction of the efficacy of ctDNA in immunotherapy for advanced gastric cancer. https://clinicaltrials.gov/ct2/show/NCT04053725 (2019).
Clinicaltrials.gov. Liquid biopsy in monitoring the neoadjuvant chemotherapy and operation in gastric cancer. https://clinicaltrials.gov/ct2/show/NCT03957564 (2019).
Clinicaltrials.gov. Radiation therapy for the treatment of metastatic gastrointestinal cancers. https://www.clinicaltrials.gov/ct2/show/NCT04221893 (2019).
We would like to acknowledge K. Kulbicki for help with sample management from the Leicester biobank, and L. Primrose, J. Riley and V. Parmar for assistance with sample processing.
Ethics approval and consent to participate
Patients were recruited in accordance with the declaration of Helsinki and consented to sample storage and use at the University of Leicester Cancer Research Biobank, Research and development project number UHL11274. Ethical approval for the project was via the East Midlands Local Research Ethics Committee (REC: 13/EM/0196).
Consent for publication
Consent for publication is included within the ethics approval from the East Midlands Local Research Ethics Committee (REC: 13/EM/0196). All patients consented to the Leicester biobank study UHL11274 consented for use of their data in publication.
Data are available in supplementary data files attached. Sequencing data readouts can be requested from the corresponding author.
The authors declare no competing interests.
This study was funded by a ‘Hope Against Cancer’, Leicester UK grant [RM60G0494] and in conjunction with the UK Department of Health on an Experimental Cancer Medicine Centre grant [C10604/A25151]. A.A.M., K.P. and D.F.G. are funded by a Cancer Research UK via programme grant to J.A.S. [C63713/A25951]. A.A.M. was also funded by a National Institute for Health Research (NIHR) Academic Clinical Fellowship for this research project.
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Openshaw, M.R., Mohamed, A.A., Ottolini, B. et al. Longitudinal monitoring of circulating tumour DNA improves prognostication and relapse detection in gastroesophageal adenocarcinoma. Br J Cancer (2020). https://doi.org/10.1038/s41416-020-1002-8