Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
Clinical Trial
. 2018 Nov;67(11):1942-1949.
doi: 10.1136/gutjnl-2017-314026. Epub 2017 Oct 30.

Methylation panel is a diagnostic biomarker for Barrett's oesophagus in endoscopic biopsies and non-endoscopic cytology specimens

Hamza Chettouh  1 , Oliver Mowforth  1 , Núria Galeano-Dalmau  1 , Navya Bezawada  1 , Caryn Ross-Innes  1 , Shona MacRae  1 , Irene Debiram-Beecham  1 , Maria O'Donovan  2 , Rebecca C Fitzgerald  1
Affiliations
Clinical Trial

Methylation panel is a diagnostic biomarker for Barrett's oesophagus in endoscopic biopsies and non-endoscopic cytology specimens

Hamza Chettouh et al. Gut. 2018 Nov.

Abstract

Objective: Barrett's oesophagus is a premalignant condition that occurs in the context of gastro-oesophageal reflux. However, most Barrett's cases are undiagnosed because of reliance on endoscopy. We have developed a non-endoscopic tool: the Cytosponge, which when combined with trefoil factor 3 immunohistochemistry, can diagnose Barrett's oesophagus. We investigated whether a quantitative methylation test that is not reliant on histopathological analysis could be used to diagnose Barrett's oesophagus.

Design: Differentially methylated genes between Barrett's and normal squamous oesophageal biopsies were identified from whole methylome data and confirmed using MethyLight PCR in biopsy samples of squamous oesophagus, gastric cardia and Barrett's oesophagus. Selected genes were then tested on Cytosponge BEST2 trial samples comprising a pilot cohort (n=20 cases, n=10 controls) and a validation cohort (n=149 cases, n=129 controls).

Results: Eighteen genes were differentially methylated in patients with Barrett'soesophagus compared with squamous controls. Hypermethylation of TFPI2, TWIST1, ZNF345 and ZNF569 was confirmed in Barrett's biopsies compared with biopsies from squamous oesophagus and gastric cardia (p<0.05). When tested in Cytosponge samples, these four genes were hypermethylated in patients with Barrett's oesophagus compared with patients with reflux symptoms (p<0.001). The optimum biomarker to diagnose Barrett's oesophagus was TFPI2 with a sensitivity and specificity of 82.2% and 95.7%, respectively.

Conclusion: TFPI2, TWIST1, ZNF345 and ZNF569 methylation have promise as diagnostic biomarkers for Barrett's oesophagus when used in combination with a simple and cost effective non-endoscopic cell collection device.

Keywords: barrett’s; biomarker; cytosponge; hypermethylation; non-endoscopic cell sampling; reflux.

PubMed Disclaimer

Conflict of interest statement

Competing interests: Since this study was conducted, the Cytosponge™-TFF3 technology has been licensed to Covidien GI solutions (now owned by Medtronic) by the Medical Research Council. Rebecca Fitzgerald and Maria O’Donovan are named inventors on patents pertaining to the Cytosponge™. Covidien Solutions and Medtronic have not been privy to this manuscript or the data therein. All other authors declare no conflicts of interest.

Figures

Figure 1
Figure 1. Testing methylation level of gene candidates in a biopsy cohort.
The methylation level of 13 genes was assessed by Methylight PCR in 5 Squamous, 7 Cardia and 9 Barretts biopsies. 10 out of 13 genes where significantly hypermethylated in Barrett’s biopsies compared to normal squamous tissue (* p<0.05; ** p>0.01). Only TFPI2, TWIST1, ZNF345 and ZNF569 were specifically hypermethylated in Barrett’s samples and not in Squamous tissue nor Cardia biopsies.
Figure 2
Figure 2. Test and validation of the 4 gene candidates in a Cytosponge− cohort.
TFPI2, TWIST1, ZNF345 and ZNF569 were significantly hypermethylated in a pilot Cytosponge− cohort (A) from Barrett’s patients compared to those from control patients (*** p<0.0001). The pilot cohort included 20 Barrett’s and 10 controls. Results were confirmed in a larger Cytosponge− cohort from the BEST2 study (B) (*** p<0.0001). The validation cohort had 149 Barrett’s and 129 controls. (C) ROC curves for selected methylated genes in the validation Cytosponge− cohort using thresholds identified in the pilot cohort.
Figure 3
Figure 3. Correlation of gene methylation status and patient age and Barrett’s segment length.
Dotplot showing correlation between the 4 methylation biomarkers (TFPI2, TWIST1, ZNF345, ZNF569) and age, circumferential (C) and maximal (M) Barrett’s lengths. The r- value represents the Pearson correlation coefficient for each variable.
See this image and copyright information in PMC

References

    1. Group MRCOCW. Surgical resection with or without preoperative chemotherapy in oesophageal cancer: a randomised controlled trial. Lancet (London, England) 2002;359(9319):1727–33. doi: 10.1016/s0140-6736(02)08651-8. [published Online First: 2002/06/07] - DOI - PubMed
    1. Lao-Sirieix P, Fitzgerald RC. Screening for oesophageal cancer. Nat Rev Clin Oncol. 2012;9(5):278–87. doi: 10.1038/nrclinonc.2012.35. - DOI - PubMed
    1. Lepage C, Rachet B, Jooste V, et al. Continuing rapid increase in esophageal adenocarcinoma in England and Wales. Am J Gastroenterol. 2008;103(11):2694–9. doi: 10.1111/j.1572-0241.2008.02191.x. - DOI - PubMed
    1. Pohl H, Sirovich B, Welch HG. Esophageal adenocarcinoma incidence: are we reaching the peak? Cancer Epidemiol Biomarkers Prev. 2010;19(6):1468–70. doi: 10.1158/1055-9965.EPI-10-0012. - DOI - PubMed
    1. Steevens J, Botterweck AA, Dirx MJ, et al. Trends in incidence of oesophageal and stomach cancer subtypes in Europe. Eur J Gastroenterol Hepatol. 2010;22(6):669–78. doi: 10.1097/MEG.0b013e32832ca091. - DOI - PubMed

Publication types