Normal Esophageal Mucosa Research Articles

Crohn's disease confined to the esophagus, with no involvement of the small or large bowel

Crohn's disease (CD) is a transmural inflammatory process that can affect any segment of the gastrointestinal tract. CD has a predilection for the colon and the small bowel, specifically the terminal ileum. Esophageal involvement of CD is exceedingly rare and there is a paucity of data regarding the disease phenotypes, course, complications and treatment. We report a case of an 18-year-old male patient presenting for intermittent dysphagia and epigastric pain. Upper endoscopy revealed esophageal ulcers in the middle esophagus surrounded by normal esophageal mucosa. Biopsies were taken from the ulcerated lesions. Histology revealed granulomatous esophagitis. The stomach and duodenum were unremarkable. An ileo-colonoscopy was performed and showed unremarkable mucosa with normal biopsies. The patient was started on proton pump inhibitors and prednisolone as induction therapy with significant symptomatic improvement within 15 days of therapy. The patient was then started on azathioprine 150 mg once daily as maintenance therapy with complete resolution of symptoms. Esophageal CD is an overlooked entity. This article underscores the need to rule out CD when physicians are confronted by esophageal ulcers on upper endoscopy.

Gastrointestinal dysmotility is associated with proton pump inhibitor refractory esophagitis in patients with systemic sclerosis.

Patients with systemic sclerosis present with severe gastroesophageal reflux disease, often refractory to proton-pump inhibitors (PPI) treatment. The aim of the present study was to identify factors associated with PPI-refractory esophagitis. We performed a cross-sectional study in a single-center cohort of patients diagnosed with systemic sclerosis. We included patients who underwent an esophagogastroduodenoscopy while on PPI treatment. Patients with PPI-refractory erosive esophagitis were compared with those with endoscopically normal esophageal mucosa. A total of 69 patients were included, from these, 23 patients (33%) had PPI-refractory esophagitis (Grade A, n = 11; Grade B, n = 7; Grade C, n = 2; Grade D, n = 3) and 46 (67%) had an endoscopically normal esophageal mucosa. On univariate analysis, patients with PPI-refractory esophagitis were more frequently diffuse SSc subset (43% vs 17%; p= 0.041). Evaluating gastrointestinal motility tests, neither absent esophageal contractility (39% vs 25%, p= 0.292) nor hypotensive lower esophageal sphincter (47% vs 44%, p= 0.980) were significantly associated with PPI-refractory esophagitis. Gastrointestinal dysmotility, defined as abnormal gastric emptying and/or small bowel dilated loops, was significantly associated with PPI-refractory esophagitis (66 vs 8%, p = <0.001). On a multivariate regression model to evaluate the association between motility test results adjusted for the diffuse subset, gastrointestinal dysmotility (β = 0.751, p= 0.010) was independently associated with PPI-refractory esophagitis, while absent esophageal contractility (β = 0.044, p= 0.886) or a hypotensive LES were not (β=-0.131, p= 0.663). Our findings suggest that gastric and small intestinal motor dysfunction may be an important contributor to the development of PPI-refractory esophagitis in patients with systemic sclerosis.

243. IS MUCOSAL CHANGE IN COELIAC DISEASE RESTRICTED TO SMALL BOWEL? STUDY OF OESOPHAGEAL MUCOSAL CHANGES IN CHILDREN WITH COELIAC DISEASE

Abstract Introduction Celiac disease (CeD) may cause changes throughout the gastrointestinal tract. It is not known whether the oesophageal mucosa is involved in CeD. The purpose of this prospective study was to study the mucosal changes in oesophagus in children with CeD. Materials &amp; Methods The mucosal changes in the esophagus was prospectively investigated in 126 children with CeD during 192 upper GI endoscopies performed to obtain duodenal biopsy specimens and was compared with findings in 49 children who underwent endoscopy for upper GI complaints without CeD (non-CeD patients) during one year. Children were diagnosed as cases of CeD based on the modified ESPGHAN criteria and positive serology. Duodenal histologic lesions were classified as proposed by our group. Results The mean age of CeD children (M:F::71:55) at presentation was 6.7 + 3.3 years. 54/126 (42.9%) of CeD and 23/49 (46.9%) of non-CeD patients showed normal oesophageal mucosa. The prevalence of lymphocytic oesophagitis (LyE) was higher in CeD (11.9%) than non-CeD (2.1%) patients. The prevalence of oeosinophilic oesophagitis, follicular oesophagitis, reflux oesophagitis and non-specific oesophageal mucosal changes were similar in both groups. All but one child with LyE showed classic duodenal histologic lesion. Conclusions Our findings showed that the oesophageal mucosa can be damaged in CeD patients and that the prevalence of LyE is higher with CeD. Awareness of potential coexistence of oesophageal lesions in CeD should promote optimal diagnosis of these conditions. Routine oesophageal biopsies may be warranted when investigating for CeD.

Expression of epidermal growth factor receptor (EGFR) in systemic sclerosis patients (SSc) and gastro-oesophageal reflux disease (GORD)

IntroductionSystemic sclerosis (SSc) affects the connective tissue and leads to an abnormal fibrotic process in the skin and internal organs. Epidermal Growth Factor Receptor (EGFR) is able to induce cell proliferation and differentiation, and its expression is increased in SSc patients with pulmonary artery hypertension and in skin biopsies in patients with scleroderma. To date, no data on esophageal expression of EGFR are available in SSc patients. We aimed to evaluate whether the pro-fibrogenic pathways of SSc may affect EGFR expression in the esophagus. MethodsA retrospective analysis included patients with SSc and control subjects suffering from gastroesophageal reflux symptoms. Endoscopic assessment and histopathologic analyses were performed in all subjects and the presence of microscopic esophagitis was used to distinguish patients with normal esophageal mucosa and subjects with non-erosive reflux disease. EGFR expression was measured in all subjects. ResultsA total of 35 patients with SSc were included, while the control group included 67 non-SSc patients. EGFR expression at the Z-line was higher in SSc patients than non-SSc patients in absence of microscopic esophagitis (median 65 %, IQR 56–71 % vs 42 %, IQR 37–54 %, p < 0.001). Microscopic esophagitis was found in 60 % of patients with SSc and 62.7 % of control patients, and EGFR expression was significantly higher in patients presenting microscopic esophagitis both in SSc and non-SSc patients. ConclusionThe EGFR hyperexpression may be due to SSc and/or reflux-related damage in patients with microscopic esophagitis. Further studies are warranted to answer open questions and provide a possible role of EGFR in terms of diagnosis, prognosis, and therapy.

High-dimensional single-cell proteomics analysis of esophageal squamous cell carcinoma reveals dynamic alterations of the tumor immune microenvironment after neoadjuvant therapy

BackgroundDynamic alterations of the tumor immune microenvironment in esophageal squamous cell carcinoma (ESCC) after different neoadjuvant therapies were understudied.MethodsWe used mass cytometry with a 42-antibody panel for 6 adjacent normal...

Endoscopic Management of Dysplastic Barrett's Oesophagus and Early Oesophageal Adenocarcinoma.

Barrett's oesophagus is a pathological condition whereby the normal oesophageal squamous mucosa is replaced by specialised, intestinal-type metaplasia, which is strongly linked to chronic gastro-oesophageal reflux. A correct endoscopic and histological diagnosis is pivotal in the management of Barrett's oesophagus to identify patients who are at high risk of progression to neoplasia. The presence and grade of dysplasia and the characteristics of visible lesions within the mucosa of Barrett's oesophagus are both important to guide the most appropriate endoscopic therapy. In this review, we provide an overview on the management of Barrett's oesophagus, with a particular focus on recent advances in the diagnosis and recommendations for endoscopic therapy to reduce the risk of developing oesophageal adenocarcinoma.

27. OPLAH PROTEIN EXPRESSION STRATIFIES THE PROGNOSIS OF PATIENTS WITH SQUAMOUS CELL CARCINOMA OF THE ESOPHAGUS

Abstract Background Squamous cell carcinoma (SCC) is one of the major subtypes of esophageal carcinoma, and accounts for up to 90% of esophageal carcinoma cases in eastern Asia and eastern and southern Africa and 50% in Western countries. The 5-year overall survival rate of ESCC patients who undergo curative treatment remains below 40%. The elucidation of molecular biomarkers could be of clinical importance both as prognostic factors besides the established clinical staging and as potential targets for treatment. We aimed to detect and validate the prognosticators of esophageal squamous cell carcinoma in patients who underwent radical esophagectomy. Methods We identified OPLAH as one of the differentially expressed genes between esophageal squamous cell carcinoma (ESCC) tissue and normal esophageal mucosa whose expression changes were significantly associated with a patient prognosis using The Cancer Genome Atlas data. The OPLAH protein expression in ESCC tissue and serum OPLAH protein concentrations were evaluated and analyzed for their association with clinicopathological factors. Results OPLAH mRNA was significantly overexpressed in ESCC tissue compared to normal esophageal mucosa, and patients with high OPLAH mRNA expression had a significantly poorer prognosis according to The Cancer Genome Atlas data. The high staining intensity of OPLAH protein in ESCC tissue clearly stratified patient prognosis. According to the multivariable analysis, high OPLAH protein expression was an independent prognostic factor for survival after surgery. Pre-neoadjuvant chemotherapy serum OPLAH protein concentrations were significantly associated with clinical tumor depth and node positivity and, consequently, with advanced clinical stage. The serum OPLAH protein concentration was significantly decreased by neoadjuvant chemotherapy. Conclusions OPLAH may serve as a useful prognosticator for ESCC patients.

Eosinophilic Oesophagitis in Patients Presenting with Dysphagia: A Retrospective Study from a Single Tertiary Care Centre in Saudi Arabia

Background and Study Aims: Eosinophilic oesophagitis (EoE) is an increasingly common cause of dysphagia in many Western countries, especially in young adults; however, the prevalence of EoE in Middle Eastern countries remains unknown. We aimed to estimate the prevalence of EoE in Saudi patients presenting with dysphagia. Additionally, we aimed to assess the practice pattern of physicians with respect to evaluating these patients.&#x0D; Patients and Methods: We performed aretrospective review of all adolescent and adult patients who underwent upper gastrointestinal endoscopy for the assessment of dysphagia. Demographic data and endoscopic and histologic findings were reviewed. The primary outcome was the prevalence of biopsy proven EoE, defined as 15 or more eosinophils per high power field (≥15eos/hpf) in patients presenting with dysphagia. Secondary outcomes included the frequency of endoscopic features suggestive of EoE and the physician’s judgement about the need for oesophageal biopsies to diagnose EoE.&#x0D; Results: A total of138 patients were identified and reviewed. The mean age was 50.7 years; 43.5% were males and 56.5% were female. Endoscopy revealed normal oesophageal mucosa in 52.9% of the patients. White plaques were reported in 4.3%, linear furrows in 2.9% and multiple rings (trachealization) in 2.2% of patients. Oesophageal biopsies were obtained from 20.3% of all patients to rule out EoE, this included 7.1% of those with normal appearance of the oesophageal mucosa. Only two patients (1.4%) with biopsy proven EoE were identified from this cohort; both were young adults.&#x0D; Conclusions: Failure to perform oesophageal biopsies in patients presenting with dysphagia as per international guidelines could explainthe low prevalence of EoE observed in this population. Alternatively, the prevalence of EoE may be lower than expected in the Middle Eastern countries. Large prospective studies are necessary to accurately assess the prevalence of EoE in Middle Eastern countries.

Clinical Significance and Prognostic Value of the Expression of LAG-3 and FGL1 in Esophageal Squamous Cell Carcinoma.

In this retrospective study, we analyzed the expression of lymphocyte activating gene 3 (LAG-3) and fibrinogen-like protein 1 (FGP1) mRNA and the corresponding proteins in 78 patients with esophageal squamous cell carcinoma (ESCC) to evaluate the clinical significance and prognostic value. mRNA and protein expression were analyzed by reverse transcription PCR and Western blotting, respectively. The expression of LAG-3 and FGL1 mRNA and the corresponding proteins in tumor tissues were significantly increased in comparison with the normal esophageal mucosa. The overexpression of LAG-3 significantly correlated with the content of tumor-infiltrating lymphocytes (TILs), tumor differentiation, and TNM stage. The overexpression of FGL1 also significantly correlated with TILs, TNM stage, and lymph node metastasis. Kaplan-Meier survival analysis showed that tumor diameter, TNM stage, lymph node metastasis, LAG-3 and FGL1 protein expression were related to the progression-free survival (p<0.05). Multivariate Cox regression showed that the level of FGL1 and TNM stage were independent prognostic factors of progression-free survival. We speculated that the tumor microenvironment of ESCC induces immunosuppression due to up-regulated expression of LAG-3 and FGL1 in the tumor tissues, which promotes tumor growth.

OPLAH Protein Expression Stratifies the Prognosis of Patients With Squamous Cell Carcinoma of the Esophagus.

Squamous cell carcinoma is one of the major subtypes of esophageal carcinoma, and the 5-year overall survival rate of esophageal squamous cell carcinoma patients who underwent curative treatment remains below 40%. We aimed to detect and validate the prognosticators of esophageal squamous cell carcinoma in patients who underwent radical esophagectomy. Comprehensive analysis of transcriptome and clinical data from The Cancer Genome Atlas revealed OPLAH as one of the differentially expressed genes between esophageal squamous cell carcinoma tissues and normal esophageal mucosa. OPLAH expression changes were significantly associated with a patient prognosis. OPLAH protein levels were further evaluated by immunohisto-chemistry in esophageal squamous cell carcinoma tissues (n=177) as well as in serum samples (n=54) using ELISA. OPLAH mRNA was significantly overexpressed in esophageal squamous cell carcinoma tissues compared to normal esophageal mucosa, and patients with high OPLAH mRNA expression have a significantly poorer prognosis, according to The Cancer Genome Atlas data. The high staining intensity of OPLAH protein in esophageal squamous cell carcinoma tissue clearly stratified patient prognosis. According to multivariable analysis, high OPLAH protein expression was an independent prognostic factor for survival after surgery. Pre-neoadjuvant chemotherapy serum OPLAH protein concentrations were significantly associated with clinical tumor depth and node positivity and, consequently, with advanced clinical stage. The serum OPLAH protein concentration was significantly decreased by neoadjuvant chemotherapy. OPLAH protein expression in cancerous tissue and serum may have clinical utility towards stratifying prognosis of patients with esophageal squamous cell carcinoma.

S125 Oral and Esophageal Manifestations in a Patient With Severe Ulcerative Colitis

Case: Ulcerative colitis (UC) is a chronic inflammatory disorder of the colon with a growing prevalence. Traditionally, UC is thought to affect the rectum and colon exclusively, and spare the remainder of the gastrointestinal (GI) tract. The course of illness can range from mild colitis to full blown toxic megacolon requiring complete colonic resection. While various treatment options are available, total colectomy is viewed as potentially curative since it has been shown to result in complete cessation of symptoms. We report an unusual case of UC in a patient who presented with oral and esophageal ulcers that resolved completely following total colectomy. A 26-year-old male with no significant past medical history presented with a 1-week history of severe oral ulceration, odynophagia and fevers along with a 4-month history of intermittent bloody diarrhea. Infectious workup was negative. CT scan of the abdomen and pelvis showed evidence of pancolitis concerning for inflammatory bowel disease (IBD). An upper endoscopy (UE) showed multiple clear base, non-bleeding esophageal ulcers and a colonoscopy revealed severe ulceration spanning the entire colon. Patient’s hospital course was complicated by development of toxic megacolon requiring urgent total colectomy with end ileostomy. Following surgery patient reported significant improvement in overall symptoms. Biopsies from the colon revealed evidence of mucosal inflammation with crypt abscesses and absence of granulomas. Biopsies from the esophagus revealed acute esophagitis with no evidence of viral or fungal infection. Patient was discharged with 4 weeks of pantoprazole therapy and inhaled budesonide however no treatment for IBD was started. A follow up magnetic resonance enterography was obtained which showed no evidence of small bowel inflammation. Postoperatively patient continued to have improvement in symptoms with adequate weight gain. His budesonide and pantoprazole therapy was discontinued. An UE was repeated 5 months later which showed normal esophageal mucosa and no evidence of ulceration. Biopsies from the esophagus showed normal mucosa with no signs of inflammation. An ileoscopy was performed which was unremarkable and biopsies showed no evidence of small intestinal inflammation. Patient continues to do well off therapy with plans of undergoing a proctocolectomy with ileal pouch-anal anastomosis. There have been approximately 20 case reports that describe upper gastrointestinal (GI) involvement in patients with UC. These patients tend to be young and have evidence of severe UC. Clean base esophageal ulcers were observed in majority of these cases. Significant advances have been made in the treatment of UC however total proctocolectomy with ileo-anal anastomosis remains as a last resort therapy which can alleviate all symptoms including extraintestinal manifestations in up to 58% of UC patients. We therefore describe a rare case of UC that presented with primarily upper GI manifestations and emphasize the notion that UC may not be restricted to the colon only and that a total colectomy can serve as a treatment option for resolution of these symptoms.

Pre-gastric secretory epithelium: A light, scanning and transmission electron microscopic study of an epithelial modification of the esophagus in embryonic quails.

The current study investigated epithelial modification of embryonic quail esophagus using gross examination, light microscopy, transmission and scanning electron microscopy. By semithin sections, the pre-gastric modified region had unfolded mucosa, formed epithelial flabs and pockets, and had reduced muscularis mucosae, thin muscular layer, less glandular tissue, and outer esophageal groove. Conversely, the normal esophageal mucosa was folded, had abundant glandular tissue and prominent muscularis mucosae, with two muscular layers; the outer and the inner. The modified epithelium resembled stratified squamous type that had a high affinity for PAS, methylene blue, and PAP stains. Ultra-structural features of the modified esophageal epithelium resembled stratified squamous epithelium and contained hypertrophic Keratinocytes; dark and light. Hypertrophic keratinocytes had RER organized, few ribosomes, and developed loose bundle of cytokeratin compared with squamous keratinocytes. Hypertrophic Keratinocytes synthesize two types of granules; peripherally located small electron-dense granules and large electron-lucent granules. Hypertrophic keratinocytes had peroxisomes that were identified by the crystalline core of the urate oxidase. In conclusion, epithelia modification may have secretory function. Further studies should be carried out to explain the exact function of this type of modified epithelium.

Expanding beyond endoscopy: A review of non-invasive modalities in Barrett's esophagus screening and surveillance.

Barrett’s esophagus (BE) is a condition that results from replacement of the damaged normal squamous esophageal mucosa to intestinal columnar mucosa and is the most significant predisposing factor for development of esophageal adenocarcinoma. Current guidelines recommend endoscopic evaluation for screening and surveillance based on various risk factors which has limitations such as invasiveness, availability of a trained specialist, patient logistics and cost. Trans-nasal endoscopy is a less invasive modality but still has similar limitations such as limited availability of trained specialist and costs. Non-endoscopic modalities, in comparison, require minimal intervention, can be done in an office visit and has the potential to be a more ideal choice for mass public screening and surveillance, particularly in patents at low risk for BE. These include newer generations of esophageal capsule endoscopy which provides direct visualization of BE, and tethered capsule endomicroscopy which can obtain high-resolution images of the esophagus. Various cell collection devices coupled with biomarkers have been used for BE screening. Cytosponge, in combination with TFF3, as well as EsophaCap and EsoCheck have shown promising results in various studies when used with various biomarkers. Other modalities including circulatory microRNAs and volatile organic compounds that have demonstrated favorable outcomes. Use of these cell collection methods for BE surveillance is a potential area of future research.

Evaluation of Gastroesophageal Reflux Disease in Children on the Autism Spectrum: A Study Evaluating the Tolerance and Utility of the BRAVO Wireless pH Monitoring.

Children on the autism spectrum disorder (ASD) may express pain or discomfort through stereotypic or self-injurious behaviors. Gastroesophageal reflux disease (GERD) may be challenging to diagnose in a child who is non-verbal or has impaired communication skills, diagnostic testing for GERD may be the only way to establish the diagnosis. We report our experience using the BRAVO wireless pH monitoring device for the evaluation of GERD in this patient population. Tolerance and feasibility as well as pH parameters and symptom correlation of the BRAVO pH were evaluated retrospectively in ASD children and compared it to a large cohort of non-ASD children. Only patients with studies lasting &gt;24 hours were included. A total of 172 patients were included, 27 of those were diagnosed with autism (median age 11 years, 17 male). We found no difference in age and weight between both groups but there was a male predominance in the autism group ( P = 0.007). We found no difference in the ability to complete at least 24 hours of study duration between both groups (24/27 or 89% in ASD vs 133/145 or 92% non-ASD patients, P = 0.632). We also found no difference in the median reflux index on the worst day ( P = 0.27) or the average of both days ( P = 0.75), BRAVO pH parameters and the proportion of abnormal studies between ASD and non-ASD children. When evaluating the overall symptom correlation with GER episodes, we did not find a difference between both groups, but we did find a higher symptom correlation for GER symptom during supine position in ASD children. Study was performed for behavioral indication in 11 ASD children, all had normal esophageal mucosa but 4 of those had an abnormal BRAVO pH study. No significant side effects were reported during the study, only 2 patients (1 non-ASD and 1 ASD) complained of self-limited chest pain. BRAVO wireless pH is well tolerated and feasible in evaluating GER and behavioral symptoms in ASD children and provides a reasonable alternative to standard trans-nasal pH monitoring.

Immune Infiltration Represents Potential Diagnostic and Prognostic Biomarkers for Esophageal Squamous Cell Carcinoma.

Background Immune infiltrates in the tumor microenvironment have established roles in tumor growth, invasion, and metastasis. However, the diagnostic and prognostic potential of immune cell signature in esophageal squamous cell carcinoma (ESCC) remains unclear. Results The proportions of 22 subsets of immune cells from 331 samples including 205 ESCC and 126 normal esophageal mucosa retrieved from TCGA, GEO, and GTEx databases were deciphered by CIBERSORT. Nine overlapping subsets of immune cells were identified as important features for discrimination of ESCC from normal tissue in the training cohort by LASSO and Boruta algorithms. A diagnostic immune score (DIS) developed by XGBoost showed high specificities and sensitivities in the training cohort, the internal validation cohort, and the external validation cohort (AUC: 0.999, 0.813, and 0.966, respectively). Furthermore, the prognostic immune score (PIS) was developed based on naive B cells and plasma cells using Cox proportional hazards model. The PIS, an independent prognostic predictor, classified patients with ESCC into low- and high-risk subgroups in the internal validation cohort (P = 0.038) and the external validation cohort (P = 0.022). In addition, a nomogram model comprising age, N stage, TNM stage, and PIS was constructed and performed excellent (HR = 4.17, 95% CI: 2.22-7.69, P < 0.0001) in all ESCC patients, with a time-dependent 5-year AUC of 0.745 (95% CI: 0.644 to 0.845), compared with PIS or TNM stage as a prognostic model alone. Conclusion Our DIS, PIS, and nomogram models based on infiltrated immune features may aid diagnosis and survival prediction for patients with ESCC.

Abstract 2805: Mass spectrometry detects lower frequency of routine markers and consistent overexpression of novel disease drivers in Barrett's-related esophageal cancer

Abstract Introduction: The incidence of esophageal adenocarcinoma (EAC) has increased more than any other cancer in our lifetime. Barrett’s esophagus (BE) is the only known precursor to EAC. At present, HER-2 and anti-PD-L1 therapies are FDA-approved for EAC, and off-label use of EGFR-directed therapy has been reported. Our study aims to use liquid chromatography mass spectrometry (LC/MS) to measure the expression of oncoproteins that are currently approved-drug targets in EAC and explore new promising targets for drug development and diagnostic applications for EAC and BE. Methods: EAC patients who underwent esophagectomy without chemo-radiation were identified. BE patients with subsequent progression to EAC and those without progression for &amp;gt;10 years were identified. Normal esophageal mucosa tissues with no history of progression were analyzed for baseline expression. Tissue sections were microdissected to isolate pure EAC, BE and normal mucosal cells from stroma. These biopsies were solubilized for LC/MS-based quantification of 80+ clinically relevant tumor markers including novel markers which are believed to play a role in the disease progression from BE to EAC. The expression trends of these markers were scrutinized by immunofluorescence (IF) in a subset of specimens/biomarkers to confirm mass spec results. Results: A total of 159 tissues were analyzed via LC-MS. In 55 EAC tissues - HER2, PD-L1 and EGFR were overexpressed in 16.3%, 0%, and 0% of patients, respectively. New markers DAD1 and S100P were noted to have high (7,634.0 and 27,222.6 attomoles/ug) and consistent (98.1% and 94.5%) overexpression in EAC samples. Expression of HER2, S100P and DAD1 were significantly higher in EAC compared to 23 normal esophageal mucosa specimens. There was no statistical difference in the expression of PD-L1 and EGFR between EAC and healthy mucosal samples. 45 BE biopsies progressed to EAC within a mean of 380 days, while the remaining 36 BE specimens did not progress over a 10-year period. 8 novel biomarkers were expressed at a higher level in BE tissue that progressed to cancer when compared to esophageal tissue that remained stable over 10 years. The expression trends observed via LC/MS were confirmed via IF in DAD1, S100P and two other novel markers. Discussion: LC-MS revealed low prevalence of target proteins in currently approved EAC therapies and high levels of newly discovered disease drivers that should be explored as possible therapy targets or diagnostic markers. The differential overexpression of the novel markers in progressive and non-progressive BE tissues, as well as the consistent overexpression of these markers in EAC tumors, demonstrates the potential for early detection of esophageal carcinogenesis and/or reveals potential therapeutic programs for an indication that lacks highly efficacious targeted therapy options. Citation Format: Christopher Hartley, Ajay Bansal, Catherine Hagen, Olivia Driscoll, Seyun Oh, Joe Abdo, Sumeet K. Mittal. Mass spectrometry detects lower frequency of routine markers and consistent overexpression of novel disease drivers in Barrett's-related esophageal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2805.

Quantity and Distribution of Eosinophils in Esophageal Specimens of Adults: An Iranian Population-Based Study.

Background & Objective:Eosinophils are normally found in different parts of the gastrointestinal tract and with less prevalence in the esophagus. Eosinophilic infiltration is increased as part of inflammatory reactions in various diseases. The aim of this study was to determine the count and distribution of eosinophils in esophageal specimens obtained for different causes. Methods: Endoscopy and pathology reports of esophageal specimens in Shahid Beheshti University related hospitals, Tehran, Iran, were extracted from 2016 to 2019. The prevalence of gastroesophageal reflux disease (GERD), malignancy, eosinophilic esophagitis, and asymptomatic patients were determined as the percentages of total resection and biopsy specimens. Each group was calculated and randomly selected according to the inclusion criteria. All data were analyzed statistically using SPSS software.Results:A total of 258 biopsy and resection specimens were evaluated in this study. Fourty three cases (16.7%) diagnosed as normal esophageal mucosa , 42 cases (16.3%) as non-specific esophagitis, 155 cases (60.1%) diagnosed as gastroesophageal reflux disease, 4 cases (1.6%) showed malignancy and other diagnoses were recorded for 14 cases (5.4%). The numbers of eosinophils in the epithelium and lamina propria in the normal group were 0.1±0.5 and 2.08±2.33, respectively. The eosinophil count in different groups and its relation to different histopathologic findings were diverse. Conclusion:The number of eosinophils within the lamina propria was significantly higher than those found within other layers. . The highest mean eosinophil count was observed in the epithelium and the lamina propria of cases diagnosed as GERD.

Paired box 5 increases the chemosensitivity of esophageal squamous cell cancer cells by promoting p53 signaling activity.

Background:Gene promoter methylation is a major epigenetic change in cancers, which plays critical roles in carcinogenesis. As a crucial regulator in the early stages of B-cell differentiation and embryonic neurodevelopment, the paired box 5 (PAX5) gene is downregulated by methylation in several kinds of tumors and the role of this downregulation in esophageal squamous cell carcinoma (ESCC) pathogenesis remains unclear.Methods:To elucidate the role of PAX5 in ESCC, eight ESCC cell lines, 51 primary ESCC tissue samples, and eight normal esophageal mucosa samples were studied and The Cancer Genome Atlas (TCGA) was queried. PAX5 expression was examined by reverse transcription-polymerase chain reaction and western blotting. Cell apoptosis, proliferation, and chemosensitivity were detected by flow cytometry, colony formation assays, and 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide assays in ESCC cell lines with PAX5 overexpression or silencing. Tumor xenograft models were established for in vivo verification.Results:PAX5 methylation was found in 37.3% (19/51) of primary ESCC samples, which was significantly associated with age (P = 0.007) and tumor-node-metastasis stage (P = 0.014). TCGA data analysis indicated that PAX5 expression was inversely correlated with promoter region methylation (r = −0.189, P = 0.011 for cg00464519 and r = −0.228, P = 0.002 for cg02538199). Restoration of PAX5 expression suppressed cell proliferation, promoted apoptosis, and inhibited tumor growth of ESCC cell lines, which was verified in xenografted mice. Ectopic PAX5 expression significantly increased p53 reporter luciferase activity and increased p53 messenger RNA and protein levels. A direct interaction of PAX5 with the p53 promoter region was confirmed by chromatin immunoprecipitation assays. Re-expression of PAX5 sensitized ESCC cell lines KYSE150 and KYSE30 to fluorouracil and docetaxel. Silencing of PAX5 induced resistance of KYSE450 cells to these drugs.Conclusions:As a tumor suppressor gene regulated by promoter region methylation in human ESCC, PAX5 inhibits proliferation, promotes apoptosis, and induces activation of p53 signaling. PAX5 may serve as a chemosensitive marker of ESCC.

Quantitative proteomic profiling of esophageal adenocarcinoma tumors to assess prevalence of approved targets and elucidate novel biomarkers.

343 Background: Esophageal adenocarcinoma (EAC) continues to have extremely poor prognosis despite advances in surgical and oncological regimens. Targeted and immune therapy hold promise to improve outcomes in solid tumors including EAC. At present HER-2 and anti-PD-L1 therapies are approved by the FDA for EAC. Additionally, off-label use of EGFR directed therapy has been reported. The aim of this study is to measure expression of 80+ oncoproteins in EAC using multiplexed mass spectrometry to elucidate expression of currently approved-drug targets and explore promising new targets for drug development. Methods: After Institutional Review Board approval (Mayo Clinic and KUMC), EAC patients whose tumors were resected via esophagectomy before chemotherapy and/or radiation were identified. Normal esophageal mucosa samples with no history of EAC were also retrieved for analysis. Tissue sections were microdissected to exclude stroma from adenocarcinoma cells. Isolated EAC cells were solubilized for mass spectrometry-based quantitation of more than 80 clinically relevant tumor markers. Results: 55 EAC tissues were analyzed. HER2, PD-L1 and EGFR were overexpressed only in 16.3%, 0%, and 0% of patients, respectively. New oncoproteins #2 and #6 were noted to have high (7,634.0 and 27,222.6 attomoles/ug) and consistent (98.1% and 94.5%) overexpression in EAC samples. Expression of HER2, oncoproteins #2 and #6 were significantly higher in EAC compared to normal esophageal mucosa. There was no statistical difference in the expression of PD-L1 and EGFR between EAC and normal samples. Conclusions: Mass spectrometry revealed a very low prevalence of target oncoproteins in EAC for currently used drugs. Our study potentially reveals alternate markers which are near universally present at high levels and should be further explored for targeted treatment.[Table: see text]

STLA101 assay for the detection of cancerous progression in Barrett’s esophagus: A multi-institutional study.

249 Background: Since 1975, Esophageal adenocarcinoma (EAC) incidence has increased more than any other cancer (̃700%). Pre-malignant metaplastic tissue, or Barrett's Esophagus (BE), is the only known precursor for EAC. Periodic histological assessment of endoscopic biopsy along dysplasia-carcinoma sequence is used to assess progression risk. Current surveillance protocols have clinical limitations as 80% of patients still present with stage II-IV EAC. The aim of this study is to compare novel proteomic expression patterns to distinguish stable BE patients from those who developed EAC using the STLA101 assay. Methods: After Institutional Review Board approval patients with BE/GERD were identified. Two cohorts were selected: progressors (progressed to EAC at subsequent interval of 0.5-3 years) and non–progressors (̃10 yrs of f/u surveillance without progression along dysplasia/carcinoma path). 24 normal esophageal mucosa samples with no h/o of BE were also retrieved for baseline expression levels. Tissue sections were microdissected to isolate pure BE cells from surrounding stromal cells. BE cells underwent a multi-step process where tissue was heated and digested to produce a liquid biopsy for mass spec. Isotopically labeled control peptides representing the STLA101 panel were spiked in with digested tissue samples to quantify all of the markers simultaneously using a Triple-quad mass spectrometer. Results: A total of 95 tissues were analyzed. 29 BE biopsies progressed to EAC within a mean of 380 days while the remaining 28 BE and 14 GERD specimens did not progress over a 10-yr period. All 8 biomarkers were expressed at a higher level in BE tissue that progressed to cancer when compared to BE/GERD tissue that remained stable over 10 years. The assay had a 100% technical success rate as no samples yielded “non-detectable” data points. Conclusions: The differential overexpression of the STLA101 panel on progressive and non-progressive BE tissues demonstrates the clinical utility of STLA101 as a predictive biomarker panel for early detection of esophageal carcinogenesis. Non-progressive BE tissues were similar to normal mucosa, therefore, patients whose BE tissue samples resemble this profile may receive a reduced screening cadence. On the other hand, patients with overexpression of the STLA101 panel may be eligible to receive more frequent screening or therapeutic intervention.