Barrett's Metaplasia Research Articles

MicroRNA Expression Profiling in the Histological Subtypes of Barrett's Metaplasia

OBJECTIVES:The histological definition of Barrett's esophagus (BE) is debated, particularly regarding the phenotype of its metaplastic columnar epithelium. Histologically proven intestinal metaplasia (IM) was the sine qua non condition for a diagnosis of BE but, more recently, non-intestinalized (i.e., cardiac gastric-type; GM) columnar metaplasia has been re-included in the spectrum of Barrett's histology. MicroRNAs modulate cell commitment, and are also reportedly dysregulated in Barrett's carcinogenesis. This study investigates miRNA expression in the histological spectrum of esophageal columnar metaplastic changes, specifically addressing the biological profile of GM vs. IM.METHODS:A study was performed to discover microRNA microarray in 30 matching mucosa samples obtained from 10 consecutive BE patients; for each patient, biopsy tissue samples were obtained from squamous, GM and intestinalized epithelium. Microarray findings were further validated by qRT-PCR analysis in another bioptic series of 75 mucosa samples.RESULTS:MicroRNA profiling consistently disclosed metaplasia-specific microRNA signatures. Six microRNAs were significantly dysregulated across the histological phenotypes considered; five of them (two overexpressed (hsa-miR-192; -miR-215) and three under-expressed (hsa-miR-18a* -miR-203, and -miR-205)) were progressively dysregulated in the phenotypic sequence from squamous to gastric-type, to intestinal-type mucosa samples.CONCLUSIONS:A consistent microRNA expression signature underlies both gastric- and intestinal-type esophageal metaplasia. The pattern of microRNA dysregulation suggests that GM may further progress to IM. The clinico-pathological implications of these molecular profiles prompt further study on the “personalized” cancer risk associated with each of these metaplastic transformations.

Mo1938 Differential Response of Normal Esophageal Squamous Cells to Acid and Bile Salt-Induced Injury May Predispose to Barrett's Metaplasia

Introduction: Normal esophageal squamous (NES) mucosa normally regenerates after injury from acid and bile salts in gastroesophageal reflux disease (GERD). Sometimes, Barrett's epithelium (BE), a columnar metaplasia, may develop after chronic GERD. BE is a precancerous condition leading to esophageal adenocarcinoma, that has the highest rate of increase in the US and western countries. It is critical therefore to understand the pathogenesis of BE in order to prevent it. Acid and bile salts, the main components of reflux, are known to induce DNA damage, activate cell cycle check points, and induce apoptosis in BE and NES cells in-vitro. Response of NES cells derived from GERD patients with and without BE to chronic acid and bile salts exposure may provide clues to molecular pathogenesis of BE. Aim: To compare expression of genes regulating proliferation, apoptosis, andmucosal healing in NES cells derived from GERD patients with and without BE in response to chronic acid and bile salt exposure. Methods: Telomerase-immortalized NES cell lines derived from GERD patients with (NES-B10T) and without (NES-G2T) BE and a Barrett's epithelial cell line (BAR-T) were used. Cells were exposed to 200μM glycocheno-deoxycholic acid at pH4 (B4) once daily for 5 minutes for three weeks similar to our earlier published studies on BAR-T and BEC (IJC2011). Results are the average of three experiments comparing transcript levels of genes involved in apoptosis (p53, Bcl2, Bax, caspase3), proliferation (Ki-67, p16) and mucosal healing (TFF3) using quantitative RT-PCR analyses. Results: In response to 3 weeks of B4 exposure, the NES-B10T and BAR-T cells induced anti apoptotic Bcl-2 but not p53 and caspase 3 mRNA expression (Table). In contrast, the NES-G2T cells induced p53, caspase3 and Bax mRNAexpression (Table). Expression of Ki-67 mRNA was induced in all the cell lines, albeit to differing levels (Table). The mRNA expression of the anti-proliferative p16 gene was induced only in NES-G2T cells. Expression of TFF3mRNAwas induced inNESG2T, undetectable inNES-B10T, andwas suppressed in BAR-T cells (Table). Conclusion: The esophageal squamous cells derived from a GERD patient without BE induce pro-apoptotic, anti-proliferative and mucosal healing genes in response to chronic acid and bile salts exposure. In contrast, esophageal squamous cells derived from a GERD patient with BE have compromised expression of genes related to mucosal healing, and induction of antiapoptotic and pro-proliferative genes. We speculate that the nature of the molecular response of NES cells to GERD may determine which GERD patients develops Barrett's metaplasia. Fold changes in gene expression compared to paired untreated cells

Mo1937 MicroRNA-192 Promotes Transition of Squamous to Columnar Phenotype: Novel Insights Into the Pathogenesis of Barrett's Esophagus

Introduction: The gastroesophageal reflux of acid and bile can cause esophagitis, and inflammation can trigger epithelial-mesenchymal transition (EMT), the process in which epithelial cells acquire phenotypic features of mesenchymal cells. EMT also occurs during carcinogenesis. Chronic exposure to acid and bile salts has been reported to cause non-neoplastic Barrett's epithelial cells in vitro to undergo malignant transformation and acquire EMT characteristics. Noting that these EMT changes occurred in cultures of initially-benign epithelial cells devoid of inflammatory cells, we hypothesized that acid and bile alone (in the absence of inflammation and neoplasia) might trigger EMT in Barrett's metaplasia. To explore this issue, we assessed features of EMT in non-neoplastic and dysplastic Barrett's epithelial cells, and we studied the effects of acid and bile salts on EMT in non-neoplastic Barrett's cells. Methods: We used: a) 2 telomerase-immortalized, non-neoplastic Barrett's epithelial cell lines (BAR-T and BAR-T10); b) BAR-T cells treated for 20 weeks with intermittent exposure to acidic bile salts (BEC-20W), and c) 3 immortalized, dysplastic Barrett's cell lines (CPB, CPC and CPD). BAR-T cells were exposed to acidic media (pH 5.5) containing conjugated bile acids (400 μM) either once for 10-minutes or for 10 minutes TID for 3 days. We evaluated features of EMT using optic morphology, Western blot and immunofluorescence (IF) for E-cadherin (CDH1) and fibronectin1 (FN1) proteins, and cell migration assays. Results: Dysplastic and BEC-20W cells had lower levels of CDH1 (an epithelial cell marker) and higher levels of FN1 (a mesenchymal cell marker) than non-neoplastic BART cells. By IF, dysplastic and BEC-20W cells demonstrated lower levels of membranous CDH1 and higher levels of cytoplasmic FN1 than BAR-T cells. Dysplastic and BEC-20W cells had a spindle-shaped appearance by optic morphology, and exhibited significantly higher migration rates than BAR-T cells (Table). In BAR-T cells, a single 10-minute exposure to acidic bile salts decreased CDH1 and increased FN1 protein levels by 6 hours. Exposure to acidic bile salts for 3 days significantly increased migration rates of BAR-T (21.7±3.3 SEM to 45.1±3.6) and BAR-T10 cells (13.2±1.8 to 22.6±2.5). Conclusion: Non-dysplastic Barrett's epithelial cells exposed to acidic bile salts for 20 weeks show evidence of EMT, as do dysplastic Barrett's epithelial cells. In non-neoplastic Barrett's cells, brief exposures to acidic bile salts induce EMT features including decreased CDH1 and increased FN1 levels, and increased cell migration. These data suggest that the gastroesophageal reflux of acid and bile might induce EMT in Barrett's metaplasia, even in the absence of inflammation and neoplasia. Conceivably, reflux-induced EMTmight contribute to remodeling and carcinogenesis in Barrett's esophagus. Migration Rates (average cells ± SEM per ≤14 fields)

Mo1912 Wide Area Transepithelial Sampling (Wats3D) Detects Barrett's Metaplasia Missed by Forceps Biopsies After Ablation of Short and Long Segment Disease

Mo1912 Wide Area Transepithelial Sampling (Wats3D) Detects Barrett's Metaplasia Missed by Forceps Biopsies After Ablation of Short and Long Segment Disease

990 Nitric Oxide Profoundly Decreases Sox-2 Expression and AKT Signaling in Esophageal Squamous Cells: A Potential Mechanism Underlying Barrett's Metaplasia

990 Nitric Oxide Profoundly Decreases Sox-2 Expression and AKT Signaling in Esophageal Squamous Cells: A Potential Mechanism Underlying Barrett's Metaplasia

Abstract 2734: Residual embryonic cells as precursors of a Barrett's-like metaplasia.

Abstract Barrett's esophagus is an intestine-like metaplasia and precursor of esophageal adenocarcinoma. Trig- gered by gastroesophageal reflux disease, the origin of this metaplasia remains unknown. p63-deficient mice, which lack squamous epithelia, may model acid-reflux damage. We show here that p63 null embryos rapidly develop intestine-like metaplasia with gene expression profiles similar to Barrett's metaplasia. We track its source to a unique embry- onic epithelium that is normally undermined and re- placed by p63-expressing cells. Significantly, we show that a discrete population of these embryonic cells persists in adult mice and humans at the squa- mocolumnar junction, the source of Barrett's meta- plasia. We show that upon programmed damage to the squamous epithelium, these embryonic cells migrate toward adjacent, specialized squamous cells in a process that may recapitulate early Barrett's. Our findings suggest that certain precancerous lesions, such as Barrett's, initiate not from genetic alterations but from competitive interactions between cell lineages driven by opportunity. Citation Format: Xia Wang. Residual embryonic cells as precursors of a Barrett's-like metaplasia. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2734. doi:10.1158/1538-7445.AM2013-2734

Abstract 3379: Overexpression of CTGF is associated with chemoresistance in esophageal adenocarcinoma.

Abstract Esophageal adenocarcinoma is the most rapidly increasing solid malignancy with an overall 5-year survival of only 19%. Neoadjuvant chemoradiation is standard practice in most large centers but results in a complete pathologic response in only 21%, although this subset has an improved 5-year survival. In this study, connective tissue growth factor (CTGF), which is associated with increased cell migration, tissue invasion, and angiogenesis, was evaluated for its role in chemoresistance in esophageal adenocarcinoma. CTGF was found to be overexpressed in 33.3% (5 of 15; mean 3.1-fold) of esophageal adenocarcinomas on oligonucleotide microarray relative to Barrett's metaplasia. Elevated expression of CTGF was also found in 5/5 advanced stage IV esophageal adenocarcinomas on Western blot. Overexpression of CTGF was observed in 70.6% of chemoresistant esophageal adenocarcinomas (n=17) as determined using qRT-PCR and was significantly higher compared to chemo-naïve esophageal adenocarcinomas (n=102; p<0.0005). Stable inhibition with an shRNA against CTGF in the esophageal adenocarcinoma cell line Flo, which expresses endogenous CTGF, led to a significantly increased response to the chemotherapeutic agents 5-FU (9.4-fold increase), cisplatin (5.5-fold), and etoposide (4.4-fold) at 24 hours (p<0.005). These findings suggest that CTGF may be important in mediating chemoresistance in esophageal adenocarcinoma and that inhibition of CTGF could be useful in modulating chemoresistance to current neoadjuvant regimens in esophageal adenocarcinomas overexpressing CTGF. Citation Format: Zhuwen Wang, Lin Lin, Dafydd G. Thomas, Mark B. Orringer, Andrew C. Chang, David G. Beer, Jules Lin. Overexpression of CTGF is associated with chemoresistance in esophageal adenocarcinoma. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3379. doi:10.1158/1538-7445.AM2013-3379

Outcomes in Patients Who Have Failed Endoscopic Therapy for Dysplastic Barrett’s Metaplasia or Early Esophageal Cancer

Endoscopic therapy (ablation ± mucosal resection) for esophageal high-grade dysplasia (HGD) or intramucosal carcinoma has demonstrated promising results. Little is known about patients who have persistent or progressive disease despite endotherapy. We compared patients who had successful eradication of their disease with those in whom endotherapy failed to try to identify factors predictive of failure and outcomes after salvage therapy. We performed a single-institution retrospective review of patients treated with endotherapy from 2007 to2012. Thirty-eight patients underwent endotherapy: 28 had successful eradication of their disease and endotherapy failed in 10 patients. Patients in whom endotherapy failed were more likely to have high-grade dysplasia (HGD) on initial endoscopy, nodules or ulcers, multifocal dysplasia, and persistent nondysplastic Barrett's metaplasia. Patients in whom endotherapy failed also underwent significantly more endotherapy sessions. Seven patients had persistent dysplasia or progression to cancer, and 3 patients had complete eradication of HGD but presented with intramucosal carcinoma an average of 15 months after eradication. The 10 patients in whom endotherapy failedunderwent salvage therapy with esophagectomy (7 patients), definitive chemoradiotherapy (1 patient), and endotherapy (2 patients). Patients treated with esophagectomy were disease free at a mean of 25 months postoperatively. HGD on initial endoscopy, multifocal dysplasia, mucosal abnormalities, and failure to eradicate nondysplastic Barrett's metaplasia were associated with failure of endotherapy. Patients with these characteristics should be considered at higher risk for treatment failure, and earlier consideration should be given to esophagectomy if there is persistent, progressive, or recurrent neoplasia. Clinical outcomes are good, even after salvage therapy. Continued endoscopic surveillance is mandatory after successful endotherapy because of the risk of recurrent disease.

MicroRNA Expression Signatures during Malignant Progression from Barrett's Esophagus to Esophageal Adenocarcinoma

Barrett's esophagus is the precursor lesion of esophageal adenocarcinoma, whose progression follows sequential stages. However, the low progression rate and the inadequacy and subjective interpretation of histologic grading in predicting Barrett's esophagus progression call for more objective biomarkers that can improve risk prediction. We conducted a genome-wide profiling of 754 human microRNAs (miRNA) in 35 normal epithelium, 34 Barrett's esophagus, and 36 esophageal adenocarcinoma tissues using TaqMan real-time PCR-based profiling. Unsupervised hierarchical clustering using 294 modestly to highly expressed miRNAs showed clear clustering of two groups: normal epithelium versus Barrett's esophagus/esophageal adenocarcinoma tissues. Moreover, there was an excellent clustering of Barrett's metaplasia (without dysplasia) tissues from normal epithelium tissues. However, Barrett's esophagus tissues of different stages and esophageal adenocarcinoma tissues were interspersed. There were differentially expressed miRNAs at different stages. The majority of miRNA aberrations involved upregulation of expression in Barrett's esophagus and esophageal adenocarcinoma tissues, with the most dramatic alterations occurring at the Barrett's metaplasia stage. Known oncomiRs, such as miR-21, miR-25, and miR-223, and tumor suppressor miRNAs, including miR-205, miR-203, let-7c, and miR-133a, showed progressively altered expression from Barrett's esophagus to esophageal adenocarcinoma. We also identified a number of novel miRNAs that showed progressively altered expression, including miR-301b, miR-618, and miR-23b. The significant miRNA alterations that were exclusive to esophageal adenocarcinoma but not Barrett's esophagus included miR-375 downregulation and upregulation of five members of the miR-17-92 and its homologue clusters, which may become promising biomarkers for esophageal adenocarcinoma development.

Increased expression of endothelin-1 and endothelin receptor A in reflux esophagitis and Barrett's esophagus

Barrett's esophagus (BE) is considered a complication of the inflammation provoked by acid and bile reflux. Endothelin-1 (ET-1) expresses in various cells during inflammatory process. However, the role of ET-1 in human inflamed and uninflamed esophageal tissue is unknown. The present study aimed to examine the expression of ET-1 and its receptors in human reflux esophagitis (RE) and BE. Endoscopic biopsies of normal squamous epithelium (NSE) (n = 20), RE (n = 22), and long segment BE (n = 14) were obtained. The segmental degree of endoscopic and histopathological inflammation was graded, and immunohistochemistry and real-time quantitative polymerase chain reaction were used to determine the expression of ET-1 and endothelin receptor A (ET(A)R) and endothelin receptor B (ET(B)R). ET-1 and ET(A)R messenger RNA (mRNA) levels were higher in RE than in NSE (3.25 ± 1.78 vs. 1.10 ± 0.71, P = 0.000; 2.13 ± 1.06 vs. 1.12 ± 0.64, P = 0.001, respectively). In BE, relative ET-1 mRNA levels in the proximal segment were higher than in the distal segment (3.03 ± 1.83 vs. 1.16 ± 0.70, P = 0.004) and in normal esophageal epithelium (P = 0.002). There was no significantly difference of ET(A)R mRNA levels between the proximal segment and the distal segment (1.99 ± 1.28 vs. 1.14 ± 0.67, P = 0.072). ET(B)R mRNA expression was unaltered between the groups. Furthermore, immunohistochemistry demonstrated that ET-1 expression increased significantly in RE (51.18 ± 30.14) compared with those in NSE (21.10 ± 18.17, P = 0.000) and in distal BE segment (28.02 ± 24.92, P = 0.022). There were more ET-1 positive cells in proximal BE segment (50.07 ± 25.88) than in distal BE segment (P = 0.030) and in NSE (P = 0.001). ET-1 expression increased in a stepwise manner with the growing degree of inflammation, and there were significant differences between mild, moderate, and marked degree esophagitis (36.08 ± 27.84, 65.86 ± 11.82, 98.00 ± 8.49, P = 0.003, respectively). However, expression of receptors remained unchanged. This study demonstrates that over-expression of ET-1 and ET(A)R in esophagitis may be related to the inflammatory process. ET-1 may play a significant role in the progression of Barrett's metaplasia.

Barrett’s Metaplasia and Colonic Neoplasms: A Significant Association in a 203,534-Patient Study

The presence of an association between Barrett's metaplasia and colonic neoplasia has remained controversial. The aim of the study was to test the presence of this association, using a large national database. From a computerized database of surgical pathology reports, we selected 203,000 subjects who underwent colonoscopy and esophago-gastro-duodenoscopy with biopsy results available from both procedures. In a case-control study we compared the occurrence of Barrett's metaplasia in patients with and without various types of colonic neoplasms. Barrett's metaplasia occurred more frequently among patients with hyperplastic polyps (OR = 2.14, 95 % CI 2.02-2.27), adenomatous polyps (2.52, 2.41-2.64), advanced adenomas (2.10, 1.90-2.32), villous adenomas or adenomas with high-grade (HG) dysplasia (2.45, 2.28-2.64), and colonic adenocarcinomas (1.75, 1.39-2.22). The association between Barrett's metaplasia and colonic neoplasm applied similarly to polyps of different size, number and location within the large bowel. These types of association could also be confirmed when analyzed separately for Barrett's metaplasia characterized by low-grade or HG dysplasia, as well as esophageal adenocarcinoma. The data support the existence of a true association between Barrett's metaplasia and various types of colonic neoplasm. The association may be more interesting for its potential insights into the pathogenesis of the two disorders than its actual clinical implications.

Laparoscopic Nissen fundoplication in the treatment of Barrett's esophagus - 10 years of experience.

IntroductionBarrett's esophagus (BE) is a state in which the distal portion of esophageal mucosa becomes lined with cylindrical epithelium as a result of adaptive remodeling. It is widely accepted that the metaplastic lesions result from chronic irritation with gastric and/or duodenal contents in the course of reflux disease. For many years, research centered on the risk factors of BE and resulting adenocarcinoma. Anti-reflux operations are the only procedures which offer the possibility of treating the cause by restoring the anatomic barrier responsible for guarding against irritating effects of gastroduodenal content on the distal esophagus. Total (i.e. 360°) laparoscopic Nissen fundoplication (LNF) is considered the most effective amongst these procedures. Still, controversies related to the indications for anti-reflux surgery are frequently encountered.AimRetrospective analysis of long-term treatment outcomes in patients with BE subjected to laparoscopic Nissen fundoplication.Material and methodsThe group included 42 BE patients, amongst them 30 men and 12 women. Initially, all the patients were treated conservatively for at least 1 year. The subgroup with dysplasia was subjected to preoperative argon plasma coagulation (APC). From 1 year after surgery (laparoscopic Nissen fundoplication), control biopsy specimens were obtained from the gastroesophageal junction of all the patients.ResultsNone of the patients showed the development of esophageal adenocarcinoma during the follow-up period. Furthermore, no cases of dysplasia progression or de novo development of dysplasia were observed in the analyzed group. In the initial 12-24 months after surgery, complete regression of metaplasia was documented in 7 (31.8%) patients from group A, and a reduction in the area of Barrett's metaplasia was observed in another 7 patients (31.8%). Throughout the period of this study, persistent planoepithelial re-epithelialization was observed in 14 (70%) group B patients, i.e. in individuals with baseline dysplasia subjected to preoperative argon plasma ablation. In the remaining patients of this group, the developed changes of BE character were less advanced than at baseline.ConclusionsOur opinion is that laparoscopic Nissen fundoplication, as a result of high effectiveness, represents the method of choice in the treatment of BE in the case of patients who were qualified for surgery.

The laparoscopic nissen-hill hybrid: pilot study of a combined antireflux procedure

Laparoscopic antireflux surgery is highly effective in patients with uncomplicated gastroesophageal reflux disease (GERD). However, long-term failure rates in paraesophageal hernia (PEH) and Barrett's metaplasia (BE) are higher and warrant a more durable repair. Outcomes for the laparoscopic Nissen fundoplication (LNF) and Hill repair (LHR) are equivalent, but their anatomic components are different and may complement each other (Aye R Ann Thorac Surg, 2012). We designed and tested the feasibility and safety of an operation that combines the essential components of each repair. A prospective, phase II pilot study was performed on patients with symptomatic giant PEH hernias and/or GERD with nondysplastic Barrett's metaplasia. Pre- and postoperative esophagogastroduodenoscopy (EGD), upper gastrointestinal study (UGI), 48-hour pH testing, manometry, and three quality-of-life metrics were obtained. Twenty-four patients were enrolled in the study. Three patients did not complete the planned procedure, leaving 21 patients, including 12 with PEH, 7 with BE, and 2 with both. There were no 30-day or in-hospital mortalities. At a median follow-up of 13 (range 6.4-30.2) months, there were no reoperations or clinical recurrences. Two patients required postoperative dilation for dysphagia, with complete resolution. Mean DeMeester scores improved from 54.3 to 7.5 (p < 0.0036). Mean lower esophageal sphincter pressures (LESP) increased from 8.9 to 21.3 mmHg (p < 0.013). Mean short-term and long-term QOLRAD scores improved from 4.09 at baseline to 6.04 and 6.48 (p < 0.0001). Mean short-term and long-term GERD-HQRL scores improved from 22.9 to 7.5 and 6.9 (p < 0.03). Mean long-term Dysphagia Severity Score Index improved from 33.3 to 40.6 (p < 0.064). The combination of a Nissen plus Hill hybrid reconstruction of the gastroesophageal junction (GEJ) is technically feasible, safe, and not associated with increased side effects. Short-term clinical results in PEH and BE suggest that this may be an effective repair, supporting the value of further study.

Synthesis and Characterization of Anti-EGFR Fluorescent Nanoparticles for Optical Molecular Imaging

Molecular imaging, the visualization of molecular and cellular markers, is a promising method for detection of dysplasia and early cancer in the esophagus and can potentially be used to identify regions of interest for biopsy or tumor margins for resection. EGFR is a previously reported cell surface receptor with stepwise increases in expression during the progression from Barrett's metaplasia to adenocarcinoma. In this work, a 200 nm fluorescent nanoparticle contrast agent was synthesized for targeted imaging of EGFR through a series of surface modifications to dye-encapsulated polystyrene particles. Amino-functionalized polystyrene particles were PEGylated using a heterobifunctional PEG linker. Subsequently, thiolated M225 antibodies were conjugated to maleimide functional groups on attached PEGs for EGFR targeting. In vitro binding studies using flow cytometry demonstrated specific binding of M225-PEG-NP to EGFR-expressing cells with minimal nonspecific binding in EGFR(-) cells. Binding was shown to increase proportionally with the number of conjugated M225 antibodies. Adsorbed formulations with unmodified M225 antibodies, M225 + PEG-NP, were synthesized using the same antibody feeds used in M225-PEG-NP synthesis to determine the contribution of adsorbed antibodies to EGFR targeting. Adsorbed antibodies were less efficient at mediated nanoparticle targeting to EGFR than conjugated antibodies. Finally, M225-PEG-NP demonstrated binding to EGFR-expressing regions in human esophageal tissue sections.

Histopathological criteria for the diagnosis of esophageal barrett metaplasia, dysplasia, and adenocarcinoma, and the potential use of Phosphorylated- β- catenin S552 and Fox M1 proteins in the diagnosis of barrett dysplasia and adenocarcinoma

Histopathological criteria for the diagnosis of esophageal barrett metaplasia, dysplasia, and adenocarcinoma, and the potential use of Phosphorylated- β- catenin S552 and Fox M1 proteins in the diagnosis of barrett dysplasia and adenocarcinoma

Diagnostic correlation between the expression of the DNA repair enzymeN-methylpurine DNA glycosylase and esophageal adenocarcinoma onset: a retrospective pilot study

EAC in its early stages, when it can potentially be cured, is rarely symptomatic and is associated with high mortality rates because in part of late-stage diagnosis. Given that DNA repair is an important contributory factor of early-stage malignancy, our study focused on the expression of the base excision repair enzyme N-methylpurine DNA glycosylase (MPG) in EAC disease onset. MPG messenger RNA (mRNA) expression levels were determined using quantitative reverse transcriptase polymerase chain reaction from a maximum of 72 patient samples. Immunohistochemistry was further utilized for the detection of MPG protein, and semiquantitative analysis performed using an H-score approach was carried out on a total of 130 archival tissue samples of different esophageal pathologies. Nuclear localized MPG protein was detected in all nonmalignant tissues derived from the enterohepatic system, with H-score values of 3.9-5.5 ± 0.4-1.0. In cancerous tissues derived from the enterohepatic system, a 9.5-fold increase in the level of MPG mRNA expression was specifically observed in the malignant regions located within the esophagus region. Further analysis revealed a 9- and 14-fold increase in MPG mRNA expression in EAC tumor, node, metastasis stages II and III, respectively, suggesting MPG expression to correlate with EAC disease progression. Immunohistochemistry analysis further showed a sevenfold significant increase in MPG protein expression in EAC tissues. Intriguingly, there was a fivefold significant decrease in nuclear localized MPG protein expression in tissues derived from Barrett's esophagus and low-grade dysplasia. Such findings highlight a complex regulatory pattern governing DNA glycosylase base excision repair initiation, as normal tissue undergoes Barrett's metaplasia and later dedifferentiates to EAC. Indeed, disease-stage-specific alterations in the expression of MPG may highlight a potential role for MPG in determining EAC onset and thus potentially be of clinical relevance for early disease detection and increased patient survival.

Abstract PR-03: Aspirin mediated downregulation of Warburg kinase AKT1 in patients with Barrett's esophagus: Implications in neoplastic transformation

Abstract Background: In patients with Barrett's esophagus, chronic reflux injury and inflammation are associated with esophageal adenocarcinoma, one of the most rapidly increasing lethal cancers. Epidemiological and preclinical studies support decreased risk of esophageal adenocarcinoma with NSAID use. Gastroesophageal refluxate contents can promote carcinogenesis in Barrett's metaplasia by activating AKT, which up regulates COX-2 expression and prostaglandin biosynthesis. We recently noted that neoplastic transformation in Barrett's esophagus is associated with increased AKT expression, particularly isoform AKT1. The AIM of this study is to investigate the effect of NSAID (Aspirin) use in patients with Barrett's esophagus on AKT1 expression using frozen tissue from the randomized, phase II trial of aspirin and esomeprazole for esophageal cancer chemoprevention in Barrett's esophagus patients and to assess the functional consequences of AKT1 expression in-vitro. Methods and Results: In a subset of patients (12 of 120 patients) with Barrett's esophagus who were randomized to placebo, lower dose (81 mg) or higher dose (325 mg) Aspirin for 4 weeks, both PCR and Western blot revealed marked reduction in AKT1 expression in patients who received 325 mg of Aspirin daily compared to patients who received placebo. Our in-vitro experiments support that AKT1 expression is functionally relevant to the process of carcinogenesis in Barrett's epithelial cells. Using adenovirus to induce AKT1 expression, we show that increased AKT1 expression increases Barrett's epithelial cell growth, facilitates RAS-mediated transformation of pre-neoplastic epithelial cells and promotes colony formation. Since AKT1 is known as Warburg kinase and regulates cellular glucose uptake to support aerobic glycolysis during growth de-regulation and neoplasia, we examined the uptake of fluorescent deoxyglucose analog (2-NBDG) in Barrett's epithelial cells transduced with AKT1 adenovirus. At 48 hrs, compared to control, AKT1 overexpression resulted in increased glucose uptake both by metaplastic and neoplastic Barrett's epithelial cells. Transcriptional repression of AKT1 abrogated the 2-NBDG uptake by these cells. Finally, we did pathway specific qPCRs and noted that use of Aspirin is associated with altered expression of chromatin remodelers that could down-regulate AKT1 expression. Conclusion: Collectively, our results demonstrate that Aspirin mediated down-regulation of Warburg kinase AKT1 in patients with Barrett?s esophagus is a novel mechanism for the regulation of Barrett?s epithelial cell growth. The in-depth characterization of molecular mechanisms is currently underway. This abstract is also presented as Poster A82. Citation Format: Navtej S. Buttar, Gary W. Falk, Cathrine J. DeMars, Sonia Chowdhury, Ahmed Elebiary, Anamay Sharma, Sidhartha Chaudhry, Nathan R. Foster, Katie L. Allen Ziegler, Yvonne Romero, Norman E. Marcon, Thomas Schnell, Douglas A. Corley, Prateek Sharma, Marcia R. Cruz-Correa, Chin Hur, David E. Fleischer, Amitabh Chak, Kenneth R. DeVault, David S. Weinberg, Gary Della’Zanna, Ellen Richmond, Thomas C. Smyrk, Sumithra Mandrekar, Paul J. Limburg. Aspirin mediated downregulation of Warburg kinase AKT1 in patients with Barrett's esophagus: Implications in neoplastic transformation. [abstract]. In: Proceedings of the Eleventh Annual AACR International Conference on Frontiers in Cancer Prevention Research; 2012 Oct 16-19; Anaheim, CA. Philadelphia (PA): AACR; Cancer Prev Res 2012;5(11 Suppl):Abstract nr PR-03.

Prevalence of buried Barrett’s metaplasia in patients before and after radiofrequency ablation

Radiofrequency ablation (RFA) to treat Barrett's esophagus is increasingly accepted. Description of the etiology, natural history, and prevalence of buried Barrett's metaplasia (BBM) following RFA is limited, although BBM continues to pose a clinical dilemma. We aimed to assess the prevalence, characteristics, and eradication rate of BBM in patients with both dysplastic and nondysplastic Barrett's esophagus, treated with RFA and followed over time. The presence of Barrett's esophagus, dysplasia, and BBM, before and after RFA, was assessed by two gastrointestinal pathologists in a retrospective chart review of patients who had undergone RFA at our center and had completed appropriate follow-up. We identified 112 patients with completed treatment and no further planned RFA. In 108, no residual Barrett's esophagus was seen after RFA; 4 patients with persistent Barrett's tissue underwent surgery. Regarding BBM, 17/112 patients (15.2%) had evidence of BBM during evaluation. In 12/17 (70.5%) BBM was found during the RFA treatment, with 8 having previously undergone non-RFA therapy and RFA for Barrett's esophagus and 4 having no previous intervention. In 5/17 (29.4%), BBM was seen only after RFA monotherapy. All 17 showed no evidence of BBM at final evaluation and were classified in the complete remission group (108/112). Both Barrett's esophagus and BBM were completely eradicated in all patients with long-term follow-up after RFA. Almost half of the patients with BBM had a prior history of non-RFA therapy for Barrett's esophagus compared with 26% the non-BBM cohort. All patients with previously identified Barrett's esophagus and BBM were completely cleared of disease at final follow-up.

Barrettʼs Metaplasia and Colonic Neoplasms Coincide in Identical Patients

Purpose: The existence of an association between Barrett's metaplasia and colonic neoplasia has remained controversial. The aim of the study was to test for the presence of such an association in a large national database. Methods: Miraca Life Sciences is a centralized pathology laboratory that serves 1,500 gastroenterologists distributed throughout the United States with more than 400,000 pathology specimens processed annually. From its computerized database, we selected 203,000 subjects who underwent colonoscopy and esophago-gastro-duodenoscopy between 1.2008 and 12.2011, with biopsy results available from both procedures. In a case-control study we compared the occurrence of Barrett's metaplasia in patients with and without various types of colonic neoplasms. Odds ratios (OR) and their 95% confidence intervals (CI) were calculated to describe the strengths of the associations between individual types of colonic neoplasms and the presence of Barrett's metaplasia. Multivariate logistic regression was used to adjust the odds ratios for age, sex, and concurrence of various histopathologic characteristics. Results: Barrett's metaplasia occurred more frequently among patients with hyperplastic polyps (OR = 2.14, 95% CI 2.02-2.27), adenomatous polyps (2.52, 2.41-2.64), advanced adenomas (2.10, 1.90-2.32), villous adenomas or adenomas with high-grade dysplasia (2.45, 2.28-2.64), and colonic adenocarcinomas (1.75, 1.39-2.22). The association between Barrett's metaplasia and colonic neoplasm applied similarly to polyps of different size, number and location within the large bowel. These types of association could also be confirmed when analyzed separately for Barrett's metaplasia characterized by low-grade or high-grade dysplasia, as well as esophageal adenocarcinoma. Conclusion: The data support the existence of a true association between Barrett's metaplasia and various types of colonic neoplasm. However, this association may be more interesting for its potential insights into the pathogenesis of the two disorders than for any clinical implication with respect to screening and surveillance of Barrett's metaplasia or colonic neoplasms. Disclosure: Dr. Genta is an employee of Miraca Life Sciences, Irving, Texas. Dr. Sonnenberg has no relevant financial relationships to disclose.

Incidence of Barrettʼs Esophagus in Dyspeptic Women Compared to Gastroesophageal Reflux Disease

Purpose: Barrett's esophagus is a precancerous condition frequently associated with chronic GERD patients. Dyspepsia is a discrete group of symptoms definition of which as per ROME III criteria demarcates it from GERD. There is, however, significant overlap of symptoms in these groups. Incidence of Barrett's esophagus in dyspeptic women has never been studied. Here in we present a small study looking a dyspeptic women and incidence of Barrett's metaplasia in them. Methods: 2 consecutive year's patient data of women who underwent initial endoscopy for chronic GERD/dyspeptic symptoms was studied retrospectively. Women undergoing repeat endoscopies were excluded. Statistical analysis was done using t test and fisher's exact test where needed. Results: In Table. Limitations: Small sample size and lack of diversity in study sample.Table: [70] ResultsConclusion: There was more esophagitis in GERD group which was statistically significant as expected. Dyspeptic women were the only ones in this study to have Barrett's metaplasia, however, no statistical significance could be attributed to this. However, it calls for a larger study to examine the real association of dyspepsia with Barrett's metaplasia.