Esophageal adenocarcinoma (EAC) is thought to develop from asymptomatic Barrett's esophagus (BE) with a low annual rate of conversion. Current endoscopy surveillance of BE patients is probably not cost-effective. Previously, we discovered serum glycoprotein biomarker candidates which could discriminate BE patients from EAC. Here, we aimed to validate candidate serum glycoprotein biomarkers in independent cohorts, and to develop a biomarker candidate panel for BE surveillance. Serum glycoprotein biomarker candidates were measured in 301 serum samples collected from Australia (4 states) and the United States (1 clinic) using previously established lectin magnetic bead array (LeMBA) coupled multiple reaction monitoring mass spectrometry (MRM-MS) tier 3 assay. The area under receiver operating characteristic curve (AUROC) was calculated as a measure of discrimination, and multivariate recursive partitioning was used to formulate a multi-marker panel for BE surveillance. Complement C9 (C9), gelsolin (GSN), serum paraoxonase/arylesterase 1 (PON1) and serum paraoxonase/lactonase 3 (PON3) were validated as diagnostic glycoprotein biomarkers in lectin pull-down samples for EAC across both cohorts. A panel of 10 serum glycoprotein biomarker candidates discriminated BE patients not requiring intervention (BE± low grade dysplasia) from those requiring intervention (BE with high grade dysplasia (BE-HGD) or EAC) with an AUROC value of 0.93. Tissue expression of C9 was found to be induced in BE, dysplastic BE and EAC. In longitudinal samples from subjects that have progressed toward EAC, levels of serum C9 were significantly (p < 0.05) increased with disease progression in EPHA (erythroagglutinin from Phaseolus vulgaris) and NPL (Narcissus pseudonarcissus lectin) pull-down samples. The results confirm alteration of complement pathway glycoproteins during BE-EAC pathogenesis. Further prospective clinical validation of the confirmed biomarker candidates in a large cohort is warranted, prior to development of a first-line BE surveillance blood test.
C9, GSN, PON1, and PON3 validated as serum biomarker candidates of EAC.
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Multimarker panel with AUROC of 0.93 to aid current endoscopy surveillance of BE.
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Induction of tissue C9 in BE, dysplastic BE and EAC.
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Alteration of complement pathway glycoproteins during BE-EAC pathogenesis.
Biomarker: Diagnostic
Glycoproteins
Gastrointestinal disease
Cancer biomarker(s)
Targeted mass spectrometry
Multiple reaction monitoring
Serum/Plasma
Barrett's esophagus
Complement pathway
Esophageal adenocarcinoma
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Author contributions: A.K.S., M.A.D., and M.M.H. designed research; A.K.S., C.W., and B.A.S. performed research; A.K.S., G.H., and R.N. analyzed data; A.K.S., G.H., D.C.W., and M.M.H. wrote the paper; G.H., K.-A.L.C., B.A.S., and M.A.D. contributed new reagents/analytic tools; I.B. evaluated and scored immunohistochemistry slides; R.N. patient sample selection for the study; W.A.P., R.V.L., A.P.B., and D.I.W. contributed to sample collection; V.J. contributed to sample collection and selection; D.C.W. contributed to sample collection and selection.
Grant support: This work was supported by The University of Queensland-Ochsner Seed Fund for Collaborative Research Grant 2014, The University of Queensland Faculty of Medicine and Biomedical Science Cancer Bequest Grant 2014, and internal funding support from The University of Queensland Diamantina Institute 2015. PROBE-NET was supported by a National Health and Medical Research Council (NHMRC) Centre of Research Excellence grant (APP1040947). KALC is a recipient of NHMRC Career Development Fellowship (APP1087415). DCW is supported by an NHMRC Research Fellowship (APP1058522). The sponsors had no influence on the study design, collection, analysis, and interpretation of data.
This article contains supplemental Tables. The authors have no conflicts of interest to disclose.
