Abstract
The incidence of Barrett’s esophagus (BE)-associated esophageal adenocarcinoma (EAC) is increasing. Next-generation sequencing (NGS) provides an unprecedented opportunity to uncover genomic alterations during BE pathogenesis and progression to EAC, but treatment-naive surgical specimens are scarce. The objective of this study was to establish the feasibility of using widely available endoscopic mucosal biopsies for successful NGS, using samples obtained from a BE ‘progressor’. Paired-end whole-genome NGS was performed on the Illumina platform using libraries generated from mucosal biopsies of normal squamous epithelium (NSE), BE and EAC obtained from a patient who progressed to adenocarcinoma during endoscopic surveillance. Selective validation studies, including Sanger sequencing, immunohistochemistry and functional assays, were performed to confirm the NGS findings. NGS identified somatic nonsense mutations of AT-rich interactive domain 1A (SWI like) (ARID1A) and PPIE and an additional 37 missense mutations in BE and/or EAC, which were confirmed by Sanger sequencing. ARID1A mutations were detected in 15% (3/20) high-grade dysplasia (HGD)/EAC patients. Immunohistochemistry performed on an independent archival cohort demonstrated ARID1A protein loss in 0% (0/76), 4.9% (2/40), 14.3% (4/28), 16.0% (8/50) and 12.2% (12/98) of NSE, BE, low-grade dysplasia, HGD and EAC tissues, respectively, and was inversely associated with nuclear p53 accumulation (P=0.028). Enhanced cell growth, proliferation and invasion were observed on ARID1A knockdown in EAC cells. In addition, genes downstream of ARID1A that potentially contribute to the ARID1A knockdown phenotype were identified. Our studies establish the feasibility of using mucosal biopsies for NGS, which should enable the comparative analysis of larger ‘progressor’ versus ‘non-progressor’ cohorts. Further, we identify ARID1A as a novel tumor-suppressor gene in BE pathogenesis, reiterating the importance of aberrant chromatin in the metaplasia–dysplasia sequence.
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Acknowledgements
The work at Johns Hopkins Medical Institutions was funded by the Jerry D’Amato foundation and the NIH (1K23DK068149) and the work at WRM’s laboratory was supported by a Cancer Center Support Grant (CA045508) from the NCI and NIH Shared Instrument grant S10 RR023702-01. MMS has been sponsored by Fulbright/the Netherland America Foundation and the René Vogels Foundation.
The patient’s consent was obtained under an IRB-approved protocol.
Author contributions: MMS acquired, analyzed the data, performed statistical analysis and drafted the manuscript. SL performed the bioinformatics analysis. SL, MD, AMM, SP and EA acquired, analyzed and interpreted data, and critically revised the manuscript. JSW, MIC, FHMM and GJO provided materials and critically revised the manuscript. EAM, WRM and AM interpreted data, critically revised the manuscript, obtained funding and supervised the study. We would like to sincerely thank Conover Talbot Jr from the Institute for Basic Biomedical Sciences at Johns Hopkins School of Medicine for his help with the bioinformatics analysis of the Affymetrix Gene Expression Array data.
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WRM has participated in Illumina sponsored meetings over the past 4 years and received travel reimbursement and honoraria for presenting at these events. The remaining authors declare no conflict of interest.
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Streppel, M., Lata, S., DelaBastide, M. et al. Next-generation sequencing of endoscopic biopsies identifies ARID1A as a tumor-suppressor gene in Barrett’s esophagus. Oncogene 33, 347–357 (2014). https://doi.org/10.1038/onc.2012.586
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DOI: https://doi.org/10.1038/onc.2012.586
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