Forkhead Box F1 Induces Columnar Phenotype and Epithelial-to-mesenchymal Transition in Esophageal Squamous Cells to Initiate Barrett's Like Metaplasia

Overview

Journal Lab Invest

Specialty Pathology

Date 2021 Jan 26

PMID 33495575

Citations 1

Authors

Alok De

Jianping Zhou

Pi Liu

Manling Huang

Sumedha Gunewardena

Sharad C Mathur

Lane K Christenson

Mukut Sharma

Qiuyang Zhang

Ajay Bansal

Affiliations

Soon will be listed here.

Abstract

Multiple genome-wide association studies (GWAS) have linked Forkhead Box F1 (FOXF1) to Barrett's esophagus (BE). Understanding whether FOXF1 is involved in initiation of Barrett's metaplasia could allow FOXF1 to be used for risk stratification and for therapy. Two-dimensional cell cultures and three-dimensional organoid cultures and well-annotated human biopsies were used to determine the role of FOXF1 in BE pathogenesis. Multiple established esophageal squamous and BE cell lines were tested in gain- and loss-of-function studies. Initiation of a BE-like metaplastic change was evaluated by measuring characteristic cytokeratins and global gene expression profiling and by culturing organoids. Epithelial-mesenchymal transition (EMT) was evaluated by immunostaining for E-cadherin, vimentin and Snail, and by cell motility assay. Columnar esophageal epithelium of BE patients exhibited higher expression of FOXF1 compared to normal squamous esophageal epithelium of GERD patients (P < 0.001). Acidic bile salts induced nuclear FOXF1 in esophageal squamous cells. FOXF1 overexpression in normal esophageal squamous cells: (a) increased columnar cytokeratins and decreased squamous cytokeratins, (b) converted squamous organoids to glandular organoids, and (c) switched global gene profiles to resemble that of human BE epithelium (P = 2.1685e - 06 for upregulated genes and P = 8.3378e - 09 for downregulated genes). FOXF1 inhibition in BE cell lines led to loss of BE differentiation markers, CK7, and mucin 2. Also, FOXF1 induced EMT and promoted cell motility in normal esophageal squamous epithelial cells. FOXF1-induced genes mapped to pathways such as Cancer, Cellular Assembly and Organization, DNA Replication, Recombination, and Repair. In conclusion, FOXF1 promotes a BE-like columnar phenotype and cell motility in esophageal squamous epithelial cells, which may have a critical role in BE development. FOXF1 should be studied further as a biomarker for BE and as a target for BE treatment.

Citing Articles

Translating Molecular Biology Discoveries to Develop Targeted Cancer Interception in Barrett's Esophagus.

Samaddar S, Buckles D, Saha S, Zhang Q, Bansal A Int J Mol Sci. 2023; 24(14).

PMID: 37511077 PMC: 10379200. DOI: 10.3390/ijms241411318.

References

Dura P, van Veen E, Salomon J, Te Morsche R, Roelofs H, Kristinsson J . Barrett associated MHC and FOXF1 variants also increase esophageal carcinoma risk. Int J Cancer. 2013; 133(7):1751-5. DOI: 10.1002/ijc.28160. View

Vergouwe F, IJsselstijn H, Biermann K, Erler N, Wijnen R, Bruno M . High Prevalence of Barrett's Esophagus and Esophageal Squamous Cell Carcinoma After Repair of Esophageal Atresia. Clin Gastroenterol Hepatol. 2017; 16(4):513-521.e6. DOI: 10.1016/j.cgh.2017.11.008. View

Goldblum J . Barrett's esophagus and Barrett's-related dysplasia. Mod Pathol. 2003; 16(4):316-24. DOI: 10.1097/01.MP.0000062996.66432.12. View

Sarosi G, Brown G, Jaiswal K, Feagins L, Lee E, Crook T . Bone marrow progenitor cells contribute to esophageal regeneration and metaplasia in a rat model of Barrett's esophagus. Dis Esophagus. 2008; 21(1):43-50. DOI: 10.1111/j.1442-2050.2007.00744.x. View

van Baal J, Verbeek R, Bus P, Fassan M, Souza R, Rugge M . microRNA-145 in Barrett's oesophagus: regulating BMP4 signalling via GATA6. Gut. 2012; 62(5):664-75. DOI: 10.1136/gutjnl-2011-301061. View

Kazumori H, Ishihara S, Kinoshita Y . Roles of caudal-related homeobox gene Cdx1 in oesophageal epithelial cells in Barrett's epithelium development. Gut. 2009; 58(5):620-8. DOI: 10.1136/gut.2008.152975. View

Nehra D, Howell P, Williams C, Pye J, Beynon J . Toxic bile acids in gastro-oesophageal reflux disease: influence of gastric acidity. Gut. 1999; 44(5):598-602. PMC: 1727508. DOI: 10.1136/gut.44.5.598. View

Wang D, Clemons N, Miyashita T, Dupuy A, Zhang W, Szczepny A . Aberrant epithelial-mesenchymal Hedgehog signaling characterizes Barrett's metaplasia. Gastroenterology. 2010; 138(5):1810-22. PMC: 3422577. DOI: 10.1053/j.gastro.2010.01.048. View

Bus P, Siersema P, van Baal J . Cell culture models for studying the development of Barrett's esophagus: a systematic review. Cell Oncol (Dordr). 2012; 35(3):149-61. PMC: 3396334. DOI: 10.1007/s13402-012-0076-6. View

10.

Landis J, Koch G . The measurement of observer agreement for categorical data. Biometrics. 1977; 33(1):159-74. View

11.

Bansal A, McGregor D, Anand O, Singh M, Rao D, Cherian R . Presence or absence of intestinal metaplasia but not its burden is associated with prevalent high-grade dysplasia and cancer in Barrett's esophagus. Dis Esophagus. 2013; 27(8):751-6. DOI: 10.1111/dote.12151. View

12.

Anders S, Pyl P, Huber W . HTSeq--a Python framework to work with high-throughput sequencing data. Bioinformatics. 2014; 31(2):166-9. PMC: 4287950. DOI: 10.1093/bioinformatics/btu638. View

13.

Clemons N, Wang D, Croagh D, Tikoo A, Fennell C, Murone C . Sox9 drives columnar differentiation of esophageal squamous epithelium: a possible role in the pathogenesis of Barrett's esophagus. Am J Physiol Gastrointest Liver Physiol. 2012; 303(12):G1335-46. DOI: 10.1152/ajpgi.00291.2012. View

14.

di Pietro M, Lao-Sirieix P, Boyle S, Cassidy A, Castillo D, Saadi A . Evidence for a functional role of epigenetically regulated midcluster HOXB genes in the development of Barrett esophagus. Proc Natl Acad Sci U S A. 2012; 109(23):9077-82. PMC: 3384195. DOI: 10.1073/pnas.1116933109. View

15.

Lee D, Duensing A . What's the FOX Got to Do with the KITten? Regulating the Lineage-Specific Transcriptional Landscape in GIST. Cancer Discov. 2018; 8(2):146-149. DOI: 10.1158/2159-8290.CD-17-1370. View

16.

Zhang H, Zhang X, Chen X, Thomas D, Hormi-Carver K, Elder F . Differences in activity and phosphorylation of MAPK enzymes in esophageal squamous cells of GERD patients with and without Barrett's esophagus. Am J Physiol Gastrointest Liver Physiol. 2008; 295(3):G470-8. PMC: 2536777. DOI: 10.1152/ajpgi.90262.2008. View

17.

Wang X, Ouyang H, Yamamoto Y, Kumar P, Wei T, Dagher R . Residual embryonic cells as precursors of a Barrett's-like metaplasia. Cell. 2011; 145(7):1023-35. PMC: 3125107. DOI: 10.1016/j.cell.2011.05.026. View

18.

Jiang M, Li H, Zhang Y, Yang Y, Lu R, Liu K . Transitional basal cells at the squamous-columnar junction generate Barrett's oesophagus. Nature. 2017; 550(7677):529-533. PMC: 5831195. DOI: 10.1038/nature24269. View

19.

Quante M, Bhagat G, Abrams J, Marache F, Good P, Lee M . Bile acid and inflammation activate gastric cardia stem cells in a mouse model of Barrett-like metaplasia. Cancer Cell. 2012; 21(1):36-51. PMC: 3266546. DOI: 10.1016/j.ccr.2011.12.004. View

20.

Sato T, Stange D, Ferrante M, Vries R, van Es J, van den Brink S . Long-term expansion of epithelial organoids from human colon, adenoma, adenocarcinoma, and Barrett's epithelium. Gastroenterology. 2011; 141(5):1762-72. DOI: 10.1053/j.gastro.2011.07.050. View