Serum glycoprotein biomarker validation for esophageal adenocarcinoma and application to Barrett’s surveillance
Authors: Shah Alok K. , Hartel Gunter , Brown Ian , Winterford Clay , Na Renhua , Lê Cao Kim-Anh , Spicer Bradley A. , Dunstone Michelle , Phillips Wayne A. , Lord Reginald V. , Barbour Andrew P. , Watson David I. , Joshi Virendra , Whiteman David C. , Hill Michelle M. 
Published: March 2018

BACKGROUND & AIMS

Esophageal 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.

METHODS

Serum 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.

RESULTS

Different 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.

CONCLUSIONS

Further 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.

Abbreviations

AAL

Aleuria aurantialectin

%CV

% Co-efficient of variation

AUROC

Area under receiver operating characteristics curve

BE

Barrett’s esophagus

BE-HGD

Barrett’s esophagus with high-grade dysplasia

BE-ID

Barrett’s esophagus which is indefinite for dysplasia

BE-LGD

Barrett’s esophagus with low-grade dysplasia

BMI

Body mass index

C1QB

Complement C1q subcomponent subunit B

C2

Complement C2

C3

Complement C3

C4B

Complement C4-B

C4BPA

C4b-binding protein alpha chain

C4BPB

C4b-binding protein beta chain

C9

Complement component C9

CFB

Complement factor B

CFI

Complement factor I

CI

Confidence interval

CP

Ceruloplasmin

EAC

Esophageal adenocarcinoma

EPHA

Erythroagglutinin fromPhaseolus vulgaris

FFPE

Formalin-fixed, paraffin-embedded

GERD

Gastroesophageal reflux disease

GSN

Gelsolin

JAC

Jacalin fromArtocarpus integrifolia

LeMBA

Lectin magnetic bead array

MRM-MS

Multiple reaction monitoring-mass spectrometry

NPL

Narcissus pseudonarcissuslectin

NSE

Non-specialized epithelium

OR

Odds ratio

PGLYRP2

N-acetylmuramoyl-L-alanine amidase

PON1

Serum paraoxonase/arylesterase 1

PON3

Serum paraoxonase/lactonase 3

RBP4

Retinol-binding protein 4

SERPINA4

Kallistatin

SIS

Stable isotope-labeled internal standard