SOX2 regulates foregut squamous epithelial homeostasis and is lost during Barrett’s esophagus development
Ramon U. Jin, Yuanwei Xu, T. Mamie Lih, Yang-Zhe Huang, Toni M. Nittolo, Blake E. Sells, Olivia M. Dres, Jean S. Wang, Qing K. Li, Hui Zhang, Jason C. Mills
Ramon U. Jin, Yuanwei Xu, T. Mamie Lih, Yang-Zhe Huang, Toni M. Nittolo, Blake E. Sells, Olivia M. Dres, Jean S. Wang, Qing K. Li, Hui Zhang, Jason C. Mills
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Research Article Development Gastroenterology

SOX2 regulates foregut squamous epithelial homeostasis and is lost during Barrett’s esophagus development

Abstract

Esophageal adenocarcinoma is increasingly prevalent and is thought to arise from Barrett’s esophagus (BE), a metaplastic condition in which chronic acid and bile reflux transforms the esophageal squamous epithelium into a gastric-intestinal glandular mucosa. The molecular determinants driving this metaplasia are poorly understood. We developed a human BE organoid biobank that recapitulates BE’s molecular heterogeneity. Bulk and single-cell transcriptomics, supported by patient tissue analysis, revealed that BE differentiation reflects a balance between SOX2 (foregut/esophageal) and CDX2 (hindgut/intestinal) transcription factors. Using squamous-specific inducible Sox2-KO (Krt5CreER/+ Sox2Δ/Δ ROSA26tdTomato/+) mice, we observed increased basal proliferation, reduced squamous differentiation, and expanded metaplastic glands at the squamocolumnar junction, some tracing back to Krt5-expressing cells. CUT&RUN analysis showed SOX2 bound and promoted differentiation-associated targets (e.g., Krt13) and repressed proliferation-associated targets (e.g., Mki67). Thus, SOX2 is critical for foregut squamous epithelial differentiation, and its decreased expression is likely an initiating step in progression to BE and then to esophageal adenocarcinoma.

Authors

Ramon U. Jin, Yuanwei Xu, T. Mamie Lih, Yang-Zhe Huang, Toni M. Nittolo, Blake E. Sells, Olivia M. Dres, Jean S. Wang, Qing K. Li, Hui Zhang, Jason C. Mills

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Figure 1

Barrett’s esophagus heterogeneity correlates with SOX2 abundance.

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Barrett’s esophagus heterogeneity correlates with SOX2 abundance. (A) Hu...
(A) Human BE samples and normal esophagus stained for SOX2, CDX2, and Alcian blue-periodic acid-Schiff (AB-PAS). Scale bars: 1 mm. (B) Patient-derived BE organoids and paired biopsies stained with H&E; organoids also stained with Alcian blue to highlight mucus cells. Scale bars: 100 μm. (C) Gene set enrichment analysis showing enrichment of “WANG-BARRETTS_ESOPHAGUS_UP” and “WANG_BARRETTS_ESOPHAGUS_DN” gene sets for esophageal squamous (SQM) organoids versus BE organoids. Normalized enrichment scores and P values shown. (D) SOX2 and CDX2 expression in 12 BE and 4 SQM organoids; Pearson’s correlation coefficient (r) and P value shown. Groupings: hindgut (blue), transitional (green), and foregut (red). (E) BE organoids stained for SOX2 (left) and CDX2 (right), categorized as high, intermediate, and low expression. Scale bars: 100 μm. (F) scRNA-Seq of BE organoid lines WU002, WU014, WU010, and WU012. Uniform manifold approximation and projection (UMAP) shows total cells; dot plots display average expression and percentage of expressing cells for SOX2, CDX2, and lineage markers (intestinal, esophageal, gastric, BE). (G) UMAPs of SOX2- and CDX2-expressing cells among organoid lines; GO biological processes for differentially expressed genes in each population listed.