Abstract 2334: Immunologic and proteomic biomarkers of progression risk in Barrett’s esophagus
Authors: Hyun Young Park, Catherine Lewis, Ashten Omstead, Erin Grayhack, Christopher Sherry, Kunhong Xiao, Gursimran Kochhar, Michael Landau, Douglas Stairs, Ali H. Zaidi, Patrick L. Wagner
Journal: Cancer Research (2025)

Abstract Barrett’s esophagus (BE) is an established risk factor for the development of high-grade dysplasia (HGD) or esophageal adenocarcinoma (EC), but clinical predictors of progression remain poorly defined. Biomarkers of progression risk are therefore in high demand, and in this study we examined the utility of immune cell density and proteomic profiling to identify progression risk indicators in biopsy samples from patients in surveillance for BE. Formalin-fixed paraffin embedded tissue samples (n=58) from patients with non-dysplastic BE undergoing surveillance biopsy were examined. During follow-up, 25 patients were diagnosed with HGD or EC (“progressors”), while 33 patients did not during at least 5 years of subsequent surveillance (“non-progressors”). To analyze immune markers for cell densities, samples were stained for CD3 and CD8 positive lymphocytes, CD68/CD86 co-positive (“M1- like”) macrophages, and CD68/CD206 co-positive (“M2-like”) macrophages. Regions of interest (ROI) were selected from the lamina propria of the gastroesophageal junction. Subgroup comparisons of cell density were carried out using the Wilcoxon rank sum test and logistic regression for the progression outcome. Proteomic profiling was assessed using label-free, liquid chromatography-tandem mass spectroscopy (LC-MS/MS), using data-independent acquisition. The raw LC-MS/MS data were searched using Spectronaut against a spectral library generated from 6 progressor and 6 non-progressor samples. Differential protein expression analysis was conducted using Perseus to compare the progressor and non-progressor groups. A total of 728 ROIs were examined for lymphocytes. lymphocyte cell densities were lower in progressors vs. non-progressors (CD3+: p=0.03; CD8+: p=0.003), as was the CD8:CD3 cell density ratio (x̄: 0.66 vs 0.84, p&amp;lt;0.0001). On logistic regression, CD8+ cell density (p=0.003) and CD8:CD3 ratio (p=0.007) were significantly associated with progression. A total of 587 and 489 ROIs were examined for M1 and M2 cell densities, respectively. M1 like macrophages were significantly increased in progressors vs non-progressors (x̄: 93.4 vs 35.4, p=0.0004). A total of 7, 209 proteins were identified across the samples, among which 163 were differentially-expressed based on progression risk (false positive rate&amp;lt;0.05). 4 major GO processes and KEGG pathways were associated with progressors: rRNA processing, Wnt signaling, bacterial invasion of epithelial cells, and G protein-coupled receptor signaling. In this study, we present immunologic and proteomic biomarkers of progression risk in BE. More comprehensive assays for immune microenvironment and mechanistic studies are planned in a prospective BE surveillance cohort obtained from an ongoing registry-repository study. The enhanced understanding of malignant progression in BE could provide opportunities to improve surveillance or prevent progression in the future. Citation Format: Hyun Young Park, Catherine Lewis, Ashten Omstead, Erin Grayhack, Christopher Sherry, Kunhong Xiao, Gursimran Kochhar, Michael Landau, Douglas Stairs, Ali H. Zaidi, Patrick L. Wagner. Immunologic and proteomic biomarkers of progression risk in Barrett’s esophagus [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 2334.