Publication details

Verification of diagnosis using CDX2 gene expression in esophageal fresh frozen tissue



Year of publication 2023
Type Conference abstract
MU Faculty or unit

Faculty of Science

Description Introduction The fresh frozen tissue (FFT) is preferably used for some molecular analyses, such as metagenomics and metatranscriptomics. In general, an examination should be performed to determine whether the bioptic tissue is pathological. CDX2 is expressed by intestinal cells and is already used as a diagnostic marker for Barrett’s esophagus (BE) and esophageal adenocarcinoma (EAC). This study aims to evaluate CDX2 gene expression in esophageal FFT from patients with gastroesophageal reflux disease (GERD) and to compare findings with histopathological diagnosis of parallel samples. Methods In 23 patients with GERD, endoscopic examination was carried out and 92 esophageal and 3 duodenal biopsies were taken. Two parallel bioptic tissues were obtained from two different sites of esophageal mucosa – site with the main pathology and adjacent site with macroscopically normal mucosa. 46 paired formalin-fixed paraffin-embedded (FFPE) samples were examined by pathologist and CDX2-staining immunohistochemistry (IHC) was performed. RNA was extracted from 46 paired esophageal FFT and 3 duodenum samples (served as positive controls) using the AllPrep DNA/RNA Mini Kit. Reverse transcription was performed using High-Capacity cDNA Reverse Transcription Kit and CDX2 mRNA levels were assessed using qPCR and Taqman Gene Expression Assays. Results All BE and EAC FFPE specimens revealed immunoreactivity for CDX2, while all esophageal FFPE samples from adjacent site were IHC-negative for CDX2. The CDX2 was expressed in 91 % of esophageal FFT with the main pathology from BE patients, in 58 % of esophageal FFT with the main pathology from EAC patients, as well as in 4.3 % of esophageal FFT taken from the macroscopically normal esophageal mucosa. Conclusion This study shows that a percentage of esophageal biopsies taken for molecular analyses has different diagnosis based on the CDX2 gene expression than parallel biopsies examined histopathollogically. For further metagenome or metatranscriptome studies, there is a need to find out a suitable marker, such as CDX2 in the case of BE and EAC, and to examine whether the tissue really represents the pathological or physiological condition.
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