Serum glycoprotein biomarker validation for esophageal adenocarcinoma and application to Barrett’s surveillance

Abstract

SUMMARYBACKGROUND & AIMSEsophageal adenocarcinoma (EAC) is thought to develop from asymptomatic Barrett’s esophagus (BE) with a low annual rate of conversion. Current endoscopy surveillance for 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 panel for BE surveillance.METHODSSerum glycoprotein biomarker candidates were measured in 301 serum samples collected from Australia (4 states) and USA (1 clinic) using lectin magnetic bead array (LeMBA) coupled multiple reaction monitoring mass spectrometry (MRM-MS). The area under receiver operating characteristic curve was calculated as a measure of discrimination, and multivariate recursive partitioning was used to formulate a multi-marker panel for BE surveillance.RESULTSDifferent glycoforms of complement C9 (C9), gelsolin (GSN), serum paraoxonase/arylesterase 1 (PON1) and serum paraoxonase/lactonase 3 (PON3) were validated as diagnostic glycoprotein biomarker candidates for EAC across both cohorts. A panel of 10 serum glycoproteins accurately discriminated BE patients not requiring intervention [BE+/-low grade dysplasia] from those requiring intervention [BE with high grade dysplasia (BE-HGD) or EAC]. Tissue expression of C9 was found to be induced in BE, dysplastic BE and EAC. In longitudinal samples from subjects that have progressed towards EAC, levels of serum C9 glycoforms were increased with disease progression.CONCLUSIONSFurther prospective clinical validation of the confirmed biomarker candidates in a large cohort is warranted. A first-line BE surveillance blood test may be developed based on these findings.AbbreviationsAALAleuria aurantia lectin%CV% Co-efficient of variationAUROCArea under receiver operating characteristics curveBEBarrett’s esophagusBE-HGDBarrett’s esophagus with high-grade dysplasiaBE-IDBarrett’s esophagus which is indefinite for dysplasiaBE-LGDBarrett’s esophagus with low-grade dysplasiaBMIBody mass indexC1QBComplement C1q subcomponent subunit BC2Complement C2C3Complement C3C4BComplement C4-BC4BPAC4b-binding protein alpha chainC4BPBC4b-binding protein beta chainC9Complement component C9CFBComplement factor BCFIComplement factor ICIConfidence intervalCPCeruloplasminEACEsophageal adenocarcinomaEPHAErythroagglutinin from Phaseolus vulgarisFFPEFormalin-fixed, paraffin-embeddedGERDGastroesophageal reflux diseaseGSNGelsolinJACJacalin from Artocarpus integrifoliaLeMBALectin magnetic bead arrayMRM-MSMultiple reaction monitoring-mass spectrometryNPLNarcissus pseudonarcissus lectinNSENon-specialized epitheliumOROdds ratioPGLYRP2N-acetylmuramoyl-L-alanine amidasePON1Serum paraoxonase/arylesterase 1PON3Serum paraoxonase/lactonase 3RBP4Retinol-binding protein 4SERPINA4KallistatinSISStable isotope-labeled internal standard