DNA Ploidy Analysis with Image Cytometry on Barrett's Esophagus-related Dysplasia and Esophageal Adenocarcinoma

Abstract

*W1585 DNA Ploidy Analysis with Image Cytometry on Barrett's Esophagus-related Dysplasia and Esophageal Adenocarcinoma Chenggong Yu, Qin Huang, Michael Klein, Raj K. Goyal Background and Aims: Barrett’s esophagus is of great clinical importance to correctly grade precursor lesions histopathologically and to identify the patients with a high risk of developing invasive malignancy for early management. Regrettably, the pathological identification of early malignancy is subjective and has high inter-observer variations, and none of the molecular markers have been proven to be sensitive and specific for this purpose also. In our previous study, we found that DNA ploidy analysis was much better than histopathology for identifying premalignant lesions in the patients with Barrett’s esophagus. In the current study, we wish to expand the on-going image cytometry study using the Automated Cellular Image System (ACIS) to measure DNA content in tissue. Methods: The cases with Barrett’s esophagus or distal esophageal adenocarcinomawere retrieved from the computer files. The lesions were identified, classified, and marked on slides by two experienced pathologists. The corresponding tissue blocks were retrieved and cut at 7 mM. The sections were then Feulgen stained. DNA content was measured and analyzed with ACIS. The control nuclei of cells with DNA index (DI) between 0.90-1.10 consisted of those of benign stromal cells including endothelial cells, fibroblasts, macrophages, and large lymphocytes. DNA content histograms were classified into near diploidy/aneuploidy (DI: 1.111.30), low aneuploidy (DI: 1.31-1.90), tetraploidy (DI: 1.91-2.10), and high aneuploidy (DI: >2.11). Results: Of the 50 cases studied, 10 were gastric cardiac mucosa without dysplasia (CM), 10 with Barrett’s esophagus without dysplasia (BE), 5 with indefinite dysplasia (ID), 9 with low-grade dysplasia (LD), 8 with high-grade dysplasia (HD), and 8 with esophageal adenocarcinoma (EA). DNA ploidy status was found in various patterns and the cases with non-diploidy including aneuploidywere identified in 0/10CM, 8/10 BE, 4/5 ID, 6/9 LD, 8/8HD, and 8/8 EA cases. ThemeanDI for each disease was 1.05 for CM, 1.20 for BE, 1.38 for ID, 1.37 for LD, 1.83 for HD, and 1.97 for EA. The results indicate that nondiploidy is common in Barrett’s esophagus-related lesions and DNA image cytometry is able to identify malignant cells in the early premalignant diseases. Conclusion: DNA content image cytometry appears to be a sensitive method for identifying premalignant lesions in Barrett’s esophagus-related diseases. Unrecognized Barrett's Esophagus in Patients with Erosive Esophagitis Unveiled by Methylene Blue Chromoendoscopy Stephen N. Wong, Jose D. Sollano Jr., Melchor M. Chan, Rolando Lopez, Johnny T. Go, Victoriano Y. Lim, Carmelita D. Dalupang, Ramon E. Carpio BACKGROUND: Recent guidelines recommend surveillance for patients with Barrett’s esophagus (BE), especially if dysplasia is present. However, in the background of erosive esophagitis (EE), endoscopic recognition of BE may be difficult. Up to 50% of patients with EE have an underlying BE discovered after the EE has healed.Methylene blue (MB) is avidly taken up by goblet cells found in columnar intestinal metaplasia (CIM), a hallmark of BE, and will stain these areas blue while leaving areas without CIM unstained. Thus, utilizing MB as a staining agent in chromoendoscopy may offer an opportunity to increase the endoscopic yield for BE in patients with EE. METHODS: All patients with EE but with no endoscopic evidence of BE were recruited into the study. Once EE was identified, 15cc of 0.5% methylene blue solution (MB) was sprayed around the distal esophagus and allowed to stay for 1-2 minutes before flushing with 100-200cc tap water. Three biopsies were then taken from both the stained and the unstained esophagealmucosa. If the distal esophagealmucosa did not stain, randombiopsies from the unstained mucosa were taken. BE was confirmed if CIM was demonstrated on histopathology. Sensitivity, specificity, positive (PPV) and negative predictive values (NPV) were computed. RESULTS: A total of 51 patients (male: female; 1.4:1) were recruited into the study. Smoking and alcohol intake were more common in males (p=0.005 and p=0.023, respectively). Over half (52.1%) of the patients had symptoms>6 months prior to endoscopy. There was no difference in demographic and clinical data among patients with and without BE (p>0.05). Distribution of esophagitis severity according to the Los Angeles classification was as follows: A (n=36, 75%); B (n=11, 22.9%); C (n=1, 2.1%). MB uptake was noted in 91.7% (n=44), 38 (86.4%) of whom were confirmed to have CIM on histopathology. In 3 patients who exhibited positive dye uptake, biopsies on both the stained and unstainedmucosa revealed CIM. The sensitivity, specificity, PPV andNPVofMBchromoendoscopy for the diagnosis of BE were 92.7%, 88.2%, 86,4% and 93.8%, respectively. There was no correlation between esophagitis severity and the presence of BE (p=0.294). CONCLUSIONS: Endoscopically unrecognized BE is highly prevalent in patients with EE. MB chromoendoscopy demonstrates high sensitivity and specificity in the diagnosis ofBEand is a valuable tool in unmasking this disease in patientswithEE.