Gastroenterology. 2018 Jun 12. pii: S0016-5085(18)34643-2. doi: 10.1053/j.gastro.2018.05.050. [Epub ahead of print]
Selection and Application of Tissue microRNAs for Non-endoscopic Diagnosis of Barrett's Esophagus.
Li X1, Kleeman S1, Coburn SB2, Fumagalli C1, Perner J3, Jammula S3, Pfeiffer RM2, Orzolek L4, Hao H4, Taylor PR2, Miremadi A5, Galeano-Dalmau N1, Lao-Sirieix P1, Tennyson M1, MacRae S1, Cook MB2, Fitzgerald RC1.
BACKGROUND & AIMS:
MicroRNA (miRNA) is highly stable in biospecimens and provides tissue-specific profiles, making it a useful biomarker of carcinogenesis. We aimed to discover a set of miRNAs that could accurately discriminate Barrett's esophagus (BE) from normal esophageal tissue and to test its diagnostic accuracy when applied to samples collected by a non-invasive esophageal cell sampling device.
We analyzed miRNA expression profiles of 2 independent sets of esophageal biopsy tissues collected during endoscopy from 38 patients with BE and 26 patients with normal esophagus (controls) using Agilent microarray and Nanostring counter assays. Consistently upregulated miRNAs were quantified by real-time PCR in esophageal tissues collected by Cytosponge from patients with BE vs without BE. miRNAs were expressed from plasmids and anti-sense oligonucleotides were expressed in normal esophageal squamous (NES) cells; effects on proliferation and gene expression patterns were analyzed.
We identified 15 miRNAs that were significantly upregulated in BE vs control tissues. Of these, 11 (MIR215, MIR194, MIR 192, MIR196a, MIR199b, MIR10a, MIR145, MIR181a, MIR30a, MIR7, and MIR199a) were validated in Cytosponge samples. The miRNAs with the greatest increases in BE tissues (7.9-fold increase inexpression or more, P<.0001: MIR196a, MIR192, MIR194, and MIR215) each identified BE vs control tissues with area under the curve (AUC) values of 0.82 or more. We developed an optimized multivariable logistic regression model, based on expression levels of 6 miRNAs (MIR7, MIR30a, MIR181a, MIR192, MIR196, and MIR199a), that identified patients with BE with an AUC value of 0.89, 86.2% sensitivity, and 91.6% specificity. Expression level of MIR192, MIR196a, MIR199a, combined that of trefoil factor 3 (TFF3), identified patients with BE with an AUC of 0.93, 93.1% sensitivity, and 93.7% specificity. Hypo-methylation was observed in the promoter region of the highly upregulated cluster MIR192-MIR194. Overexpression of these miRNAs in NES cells increased their proliferation, via GRHL3 and PTEN signaling.
In analyses of miRNA expression patterns of BE vs non-BE tissues, we identified a profile that can identify Cytosponge samples from patients with BE with an AUC of 0.93. Expression of MIR194 is increased in BE samples via epigenetic mechanisms that might be involved in BE pathogenesis.
Copyright © 2018 AGA Institute. Published by Elsevier Inc. All rights reserved.
biomarker; diagnosis; esophageal adenocarcinoma; gene regulation
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