Esophageal Squamous Epithelium Research Articles

NEDD4L mediates ITGB4 ubiquitination and degradation to suppress esophageal carcinoma progression

The ubiquitination-mediated protein degradation exerts a vital role in the progression of multiple tumors. NEDD4L, which belongs to the E3 ubiquitin ligase NEDD4 family, is related to tumor genesis, metastasis and drug resistance. However, the anti-tumor role of NEDD4L in esophageal carcinoma, and the potential specific recognition substrate remain unclear. Based on public esophageal carcinoma database and clinical sample data, it was discovered in this study that the expression of NEDD4L in esophageal carcinoma was apparently lower than that in atypical hyperplastic esophageal tissue and esophageal squamous epithelium. Besides, patients with high expression of NEDD4L in esophageal carcinoma tissue had longer progression-free survival than those with low expression. Experiments in vivo and in vitro also verified that NEDD4L suppressed the growth and metastasis of esophageal carcinoma. Based on co-immunoprecipitation and proteome analysis, the NEDD4L ubiquitination-degraded protein ITGB4 was obtained. In terms of the mechanism, the HECT domain of NEDD4L specifically bound to the Galx-β domain of ITGB4, which modified the K915 site of ITGB4 in an ubiquitination manner, and promoted the ubiquitination degradation of ITGB4, thus suppressing the malignant phenotype of esophageal carcinoma.

Prognostic Significance of NQO1 Expression in Non-neoplastic Esophageal Squamous Epithelium for Patients With Esophageal Cancer.

NAD(P)H dehydrogenase [quinone] 1 (NQO1), an antioxidant enzyme, confers resistance to anticancer agents. NQO1 C609T is a single-nucleotide polymorphism associated with reduced protein expression in the non-neoplastic esophageal squamous epithelium (ESE). This study aimed to investigate immunohistochemical NQO1 expression in non-neoplastic ESE and to elucidate its prognostic significance in patients with esophageal squamous cell carcinoma (ESCC) undergoing neoadjuvant therapy followed by esophagectomy. NQO1 expression in non-neoplastic ESE was determined in surgical specimens from 83 patients with ESCC using immunohistochemistry. The association between NQO1 expression and clinicopathological factors, and the prognostic significance of NQO1 expression for relapse-free survival (RFS) were statistically evaluated. Patients with complete loss or weak NQO1 expression and patients with moderate or strong NQO1 expression were classified into the NQO1-negative (n=29) and NQO1-positive (n=54) groups, respectively. The downstaging of T classification status after neoadjuvant therapy was significantly more frequent in the NQO1-negative group than in the NQO1-positive group (59% vs. 33%; p=0.036). The NQO1-negative group had significantly more favorable RFS than the NQO1-positive group (p=0.035). Multivariate survival analysis demonstrated that NQO1 negative expression had a favorable prognostic impact on RFS (HR=0.332; 95%CI=0.136-0.812; p=0.016). Immunohistochemical evaluation of NQO1 expression in non-neoplastic ESE has clinical utility for predicting patient prognosis after neoadjuvant therapy followed by esophagectomy and might be helpful for selecting candidates for adjuvant therapy to treat ESCC.

Esophageal squamous cell carcinoma with pagetoid spread: a clinicopathologic study.

Pagetoid spread in esophageal squamous epithelium associated with underlying esophageal adenocarcinoma (EAC) has been well studied. Case reports describing pagetoid spread of esophageal squamous cell carcinomas (ESCC) also exist in the literature. The latter, however, has not been systematically studied. In this study, we report seven cases of pagetoid spread associated with ESCC. The clinical, morphologic, and immunophenotypic profiles of pagetoid spread in the context of ESCC and EAC are compared. Cases of pagetoid spread of ESCC were identified through computerized search of pathology archives at five institutions. Additional cases were identified through manual review of surgical resection cases of treatment naive ESCC in Mass General Brigham (MGB) pathology archive. Clinical history was collected via chart review. Immunohistochemistry for CK7, CK20, CDX2, p53, p63, and p40 was performed on selected cases. A computerized search of pathology archives of five institutions revealed only two cases. A manual review of 76 resected untreated ESCC revealed five additional cases with unequivocal pagetoid spread of ESCC, indicating the condition was not uncommon but rarely reported. Patient age ranged from 54 to 78years (median, 65). There were six women and one man. One case had in situ disease, five had pT1 (1 pT1a and 4 pT1b), and one had pT3 disease. One of the patients with pT1 tumor had a positive lymph node, while the remaining six patients were all N0. Four tumors were in the proximal to mid esophagus, and three in the distal esophagus. Patient survival ranged from 25months to more than 288months. The pagetoid tumor cells demonstrated enlarged, hyperchromatic nuclei with variable amounts of eosinophilic cytoplasm. The cytoplasm was often condensed to the perinuclear area, creating peripheral clearing. By immunohistochemistry, the pagetoid cells were positive for p40 (6/6) and p63 (7/7) and negative for CDX2 (7/7). The tumor cells showed mutant-type staining for p53 in five of seven cases. One of the patients had pagetoid tumor cells at the resection margin and subsequently had recurrent disease 2years later. All other patients had negative resection margins and did not have local recurrence. Four cases of pagetoid spread in the context of EAC were used as a comparison group. Previously published studies were also analyzed. These tumors were all located in the distal esophagus or gastroesophageal junction. All cases were associated with underlying invasive EAC. Pagetoid spread associated with EAC often had cytoplasmic vacuoles or mucin. They were more frequently positive for CK7 than pagetoid ESCC (p = 0.01). Both ESCC and EAC may give rise to pagetoid spread of tumor cells within surface squamous epithelium. Pagetoid spread from ESCC and EAC have overlapping morphologic features. P40 and p63 immunostains can facilitate the distinction between ESCC and EAC. P53 immunostain can aid in confirmation of malignancy. Understanding their overlapping pathologic features will help pathologists avoid pitfalls and diagnose these lesions correctly on biopsy specimens.

On-chip Raman spectroscopy of live single cells for the staging of oesophageal adenocarcinoma progression.

The absence of early diagnosis contributes to oesophageal cancer being the sixth most common cause of global cancer-associated deaths, with a 5-year survival rate of < 20%. Barrett's oesophagus is the main pre-cancerous condition to adenocarcinoma development, characterised by the morphological transition of oesophageal squamous epithelium to metaplastic columnar epithelium. Early tracking and treatment of oesophageal adenocarcinoma could dramatically improve with diagnosis and monitoring of patients with Barrett's Oesophagus. Current diagnostic methods involve invasive techniques such as endoscopies and, with only a few identified biomarkers of disease progression, the detection of oesophageal adenocarcinoma is costly and challenging. In this work, single-cell Raman spectroscopy was combined with microfluidic techniques to characterise the development of oesophageal adenocarcinoma through the progression of healthy epithelial, Barrett's oesophagus and oesophageal adenocarcinoma cell lines. Principal component analysis and linear discriminant analysis were used to classify the different stages of cancer progression. with the ability to differentiate between healthy and cancerous cells with an accuracy of 97%. Whilst the approach could also separate the dysplastic stages from healthy or cancer with high accuracy-the intra-class separation was approximately 68%. Overall, these results highlight the potential for rapid and reliable diagnostic/prognostic screening of Barrett's Oesophagus patients.

Does radiofrequency ablation of the lower oesophagus allow for clonal expansion of highly mutated neosquamous epithelium?

ObjectiveIn Barrett’s oesophagus (BE), after radiofrequency ablation (RFA), the oesophagus can be repopulated with a stratified ‘neosquamous epithelium’ (NeoSE). While histologically normal, the origin and clonal make-up of this NeoSE...

How should extra-large Lugol-unstained lesions of the esophagus be treated? Results from a population-based cohort study.

Current guidelines recommend only severe dysplasia and above (SDA) lesions of the esophageal squamous epithelium for clinical intervention. However, the histopathologic diagnosis derived from tissue biopsies may be subject to underestimation of severity. 1073 participants from whom biopsies were taken at baseline chromoendoscopic examination in a population-based screening trial were enrolled in this study. The size of the Lugol-unstained lesions (LULs) was mainly analyzed. The outcome was defined as SDA lesions either identified at baseline screening, or during follow-up, collectively referred to as the cumulative risk of SDA. Multivariable logistic regression models were used to evaluate the cumulative risk of SDA. One hundred and forty-six SDA cases were identified in the study period. Participants with large LULs had a high cumulative incidence of SDA (cumulative incidence16-20mm : 55.88%; cumulative incidence>20mm : 76.92%) in the median of 7-year duration. LULs of large size were significantly associated with a higher cumulative risk of SDA, regardless of the pathologic diagnosis (adjusted OR16-20mmvs.≤5mm = 21.02, 95% CI: 7.56-58.47; adjusted OR>20mmvs.≤5mm = 33.62, 95% CI: 11.79-95.87). Results from this study suggest physician-patient shared decision-making regarding clinical treatment or intensive surveillance should be carried out for LULs >20 mm in the esophagus, regardless of the histologic diagnosis. For those with LULs of 16-20 mm, intensive surveillance would also best be considered.

350. FUNCTIONAL RESPONSES OF SWINE ESOPHAGUS TO FOCAL CRYOTHERAPY

Abstract Barrett’s esophagus is a premalignant condition where the squamous epithelium of the distal esophagus is replaced by columnar epithelium. Treatment involves endoscopic resection followed by ablation for complete eradication. Various ablative techniques exist including cryotherapy which utilizes a cryogen spray or balloon catheter. While efficacious, complications such as dysphagia have been reported. To better understand the underlying mechanism of how cryotherapy might damage the muscularis, isolated muscle strips were ablated and their function studied. Isolated muscle strips with the mucosa layers removed were prepared from freshly isolated swine esophagus and placed in tissue baths where they were electrically stimulated, and contractile forces were recorded. Contractile responses were allowed to stabilize before a given muscle strip was removed for focal ablation. Cryoablation was performed with a focal cardiac ablation catheter (Freezor, Medtronic). Following ablation, the given muscle strip was returned to the tissue baths. Regression analyses were performed using temperature data from a thermocouple integrated into the tip of the ablation catheter and contractile forces after the esophageal muscle strips had recovered for 3 hours. Ablations lasting less than 60 seconds were applied to 135 esophageal muscle strips: with an average mass of 94 mg. From the experimental data, a multiple linear regression model was generated: 71.93 + 0.4770x—0.5530y = z, where x is the minimum temperature (C), y is the time below 0 C, and z is the contractile force (normalized to preablative force, %). Our model has an adjusted R-squared value of 0.508. As expected, lower minimum temperatures and longer treatment times were associated with greater losses in stimulated muscle forces. We were able to reproducible ablate isolated swine esophageal strips that elicited contractile responses in vitro. Lower temperatures and longer treatment times were associated with greater losses in stimulated muscle forces. Yet, the adjusted R-squared value indicates that temperature and time only partially explain the response. Future studies could be improved by accounting for variabilities in sample sizes and additionally, regression diagnostics need to be performed to check for outliers and validate model assumptions.

Atrial Flutter After Radiofrequency Ablation for Barrett's Esophagus: A Case Report.

BACKGROUND Barrett's esophagus (BE) is a metaplastic change in the normal esophageal squamous epithelium and is a well-recognized precursor of esophageal adenocarcinoma (EAC). Nowadays, focal radiofrequency ablation is a valid technique for BE treatment by inducing a superficial and focal thermic destruction of metaplastic tissues. According to the literature, the most frequent patient-related adverse events of this procedure are esophageal iatrogenic stenosis, mucosal laceration or perforation of the esophagus, chest pain, and odynophagia/dysphagia. Postoperative heart rhythm abnormalities have been reported very rarely. CASE REPORT A 74-year-old patient with HE was treated by radiofrequency ablation (RFA) with the Barrx™ catheter system. He had 2 symptomatic episodes of atrial flutter in the immediate postoperative period requiring an external electrical cardioversion to induce a return to sinus cardiac rhythm. After atrial flutter ablation, 2 more radiofrequency procedures were performed, without adverse events. A laparoscopic Nissen fundoplication was carried out with complete endoscopic and histologic eradication of BE after 12-month follow-up. To the best of our knowledge, this is the first reported case of atrial flutter after esophageal RFA. Different mechanisms acting on an anatomic predisposing substrate can potentially play a role in starting atrial flutter, and include inflammation, autonomic activation, and myocardial injury. CONCLUSIONS The occurrence of this new type of adverse effect could potentially modify indications and postoperative monitoring of RFA treatment for BE. Endoscopists should know the possibility of this procedural complication in high-risk patients and they should propose alternative techniques or implement close cardiac monitoring in the postoperative period.

IL-1β transgenic mouse model of inflammation driven esophageal and oral squamous cell carcinoma

Chronic inflammation is integral to the development of esophageal adenocarcinoma (EAC) and esophageal squamous cell carcinoma (ESCC), although the latter has not been associated with reflux esophagitis. The L2-IL-1β transgenic mice, expressing human interleukin (IL)-1β in the oral, esophageal and forestomach squamous epithelia feature chronic inflammation and a stepwise development of Barrett’s esophagus-like metaplasia, dysplasia and adenocarcinoma at the squamo-columnar junction. However, the functional consequences of IL-1β-mediated chronic inflammation in the oral and esophageal squamous epithelia remain elusive. We report for the first time that in addition to the previously described Barrett’s esophagus-like metaplasia, the L2-IL-1β mice also develop squamous epithelial dysplasia with progression to squamous cell carcinoma (SCC) in the esophagus and the tongue. L2-IL-1β showed age-dependent progression of squamous dysplasia to SCC with approximately 40% (n = 49) and 23.5% (n = 17) incidence rates for esophageal and tongue invasive SCC respectively, by 12–15 months of age. Interestingly, SCC development and progression in L2-IL-1β was similar in both Germ Free (GF) and Specific Pathogen Free (SPF) conditions. Immunohistochemistry revealed a T cell predominant inflammatory profile with enhanced expression of Ki67, Sox2 and the DNA double-strand break marker, γ-H2AX, in the dysplastic squamous epithelia of L2-IL-1β mice. Pro-inflammatory cytokines, immunomodulatory players, chemoattractants for inflammatory cells (T cells, neutrophils, eosinophils, and macrophages) and oxidative damage marker, iNOS, were significantly increased in the esophageal and tongue tissues of L2-IL-1β mice. Our recent findings have expanded the translational utility of the IL-1β mouse model to aid in further characterization of the key pathways of inflammation driven BE and EAC as well as ESCC and Oral SCC.

Blockage of retinoic acid signaling via RARγ suppressed the proliferation of pancreatic cancer cells by arresting the cell cycle progression of the G1-S phase

BackgroundOur study and several studies have reported that in some cancers, including pancreatic ductal adenocarcinoma (PDAC), the expression of squamous lineage markers, such as esophagus-tissue-specific genes, correlated with a poor prognosis. However, the mechanism by which the acquisition of squamous lineage phenotypes leads to a poor prognosis remains unclear. We previously reported that retinoic acid signaling via retinoic acid receptor γ (RARγ signaling) determines the differentiation lineage into the esophageal squamous epithelium. These findings hypothesized that the activation of RARγ signaling contributed to acquiring squamous lineage phenotypes and malignant behavior in PDAC.MethodsThis study utilized public databases and immunostaining of surgical specimens to examine RARγ expression in PDAC. We evaluated the function of RARγ signaling by inhibitors and siRNA knockdown using a PDAC cell line and patient-derived PDAC organoids. The mechanism of the tumor-suppressive effects by blocking RARγ signaling was examined by a cell cycle analysis, apoptosis assays, RNA sequencing and Western blotting.ResultsRARγ expression in pancreatic intraepithelial neoplasia (PanIN) and PDAC was higher than that in the normal pancreatic duct. Its expression correlated with a poor patient prognosis in PDAC. In PDAC cell lines, blockade of RARγ signaling suppressed cell proliferation by inducing cell cycle arrest in the G1 phase without causing apoptosis. We demonstrated that blocking RARγ signaling upregulated p21 and p27 and downregulated many cell cycle genes, including cyclin-dependent kinase 2 (CDK2), CDK4 and CDK6. Furthermore, using patient-derived PDAC organoids, we confirmed the tumor-suppressive effect of RARγ inhibition and indicated the synergistic effects of RARγ inhibition with gemcitabine.ConclusionsThis study clarified the function of RARγ signaling in PDAC progression and demonstrated the tumor-suppressive effect of selective blockade of RARγ signaling against PDAC. These results suggest that RARγ signaling might be a new therapeutic target for PDAC.

A screened predictive model for esophageal squamous cell carcinoma based on salivary flora data.

Esophageal squamous cell carcinoma (ESCC) is a malignant tumor of the digestive system in the esophageal squamous epithelium. Many studies have linked esophageal cancer (EC) to the imbalance of oral microecology. In this work, different machine learning (ML) models including Random Forest (RF), Gaussian mixture model (GMM), K-nearest neighbor (KNN), logistic regression (LR), support vector machine (SVM) and extreme gradient boosting (XGBoost) based on Genetic Algorithm (GA) optimization was developed to predict the relationship between salivary flora and ESCC by combining the relative abundance data of Bacteroides, Firmicutes, Proteobacteria, Fusobacteria and Actinobacteria in the saliva of patients with ESCC and healthy control. The results showed that the XGBoost model without parameter optimization performed best on the entire dataset for ESCC diagnosis by cross-validation (Accuracy = 73.50%). Accuracy and the other evaluation indicators, including Precision, Recall, F1-score and the area under curve (AUC) of the receiver operating characteristic (ROC), revealed XGBoost optimized by the GA (GA-XGBoost) achieved the best outcome on the testing set (Accuracy = 89.88%, Precision = 89.43%, Recall = 90.75%, F1-score = 90.09%, AUC = 0.97). The predictive ability of GA-XGBoost was validated in phylum-level salivary microbiota data from ESCC patients and controls in an external cohort. The results obtained in this validation (Accuracy = 70.60%, Precision = 46.00%, Recall = 90.55%, F1-score = 61.01%) illustrate the reliability of the predictive performance of the model. The feature importance rankings obtained by XGBoost indicate that Bacteroides and Actinobacteria are the two most important factors in predicting ESCC. Based on these results, GA-XGBoost can predict and diagnose ESCC according to the relative abundance of salivary flora, providing an effective tool for the non-invasive prediction of esophageal malignancies.

Abstract A013: Induced pluripotent stem cells derived from normal human esophageal epithelial cells identify transcription factor networks contributing to esophageal carcinogenesis

Abstract Esophageal cancer (EsC) is the sixth leading cause of cancer-related mortality worldwide. Esophageal squamous cell cancers (ESCC) typically arise in the upper and mid-esophagus, whereas esophageal adenocarcinomas (EAC) arise in the distal esophagus and gastroesophageal junction. Presently, epigenetic mechanisms contributing to initiation, progression, and dissemination of EsC have not been fully elucidated. The present study was undertaken to determine if induced pluripotent stem cells (iPSCs) derived from normal esophageal epithelial cells (Eso-iPSCs), EsC cell lines cultured in stemness enriched conditions, and cancer stem-like cells isolated from primary EsC specimens could be used to identify unique primitive stem-like transcription factor networks associated with epigenetic plasticity, chemoresistance, and metastatic potential of EsC cells. RNA-seq analysis demonstrated that genes differentially expressed in Eso-iPSC overlapped with transcriptome signatures in lung-iPSC (Lu-iPSC) which we previously generated to identify novel therapeutic targets in lung cancers; similar to Lu-iPSC, Eso-iPSC transcriptome signatures overlapped considerably more with small cell lung cancers (SCLC) compared to non-small cell lung cancers (NSCLC), possibly reflecting greater stemness in SCLC. We next compared transcriptome signatures of Eso-iPSC with stem-like-EAC enrichment models (cell lines and PDX). 6% and 8% of differentially expressed genes in Eso-iPSC uniquely overlapped with EAC or ESCC lines, respectively, whereas 24% were common to Eso-iPSC, EAC cells, and ESCC cells. Culturing of EAC cells on matrigel or propagation of EAC as primary patient derived xenografts increased commonalities of Eso-iPSC and EAC transcriptome signatures. Lastly, we compared transcriptomes of Eso-iPSC with Barrett’s esophagus (BE; a premalignant condition in which the squamous epithelium of the distal esophagus is replaced by metaplastic columnar epithelial cells) and EAC samples. By combining and stringently filtering transcriptome, ATAC-seq, DNA methylome, and ChIP-seq datasets for tissue- and disease-specific signatures, we identified several potentially druggable, master TFs mediating stemness/plasticity, chemoresistance, and metastatic potential which are shared between EsC lines and primary tumor specimens. Enrichment techniques promoted acquisition of more stem-like states in cultured EAC cells as reflected in multi-omic signatures of these cells relative to Eso-iPSC and primary EAC specimens. Collectively, these findings establish proof of principal for our experimental approach which is now being used to further characterize epigenetic mechanisms contributing to esophageal carcinogenesis and to identify novel targets for EsC therapy. Citation Format: Haitao Wang, Ruihong Wang, Katherine P. Prothro, Sichuan Xi, Weilong Hou, Xinwei Wu, Tuana Tolunay, Vivek Shukla, Mary R. Zhang, Stephanie J. Shiffka, Sudheer Gara, David S. Schrump. Induced pluripotent stem cells derived from normal human esophageal epithelial cells identify transcription factor networks contributing to esophageal carcinogenesis. [abstract]. In: Proceedings of the AACR Special Conference: Cancer Epigenomics; 2022 Oct 6-8; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2022;82(23 Suppl_2):Abstract nr A013.

Safety and efficacy of liquid nitrogen spray cryotherapy in Barrett's neoplasia - a comprehensive review and meta-analysis.

Background and study aims Barrett's esophagus (BE) is a precursor condition to esophageal adenocarcinoma (EAC), resulting in transformation of the squamous epithelium of distal esophagus to columnar-lined epithelium with intestinal metaplasia (IM). Liquid nitrogen spray cryotherapy (LNSC) is a non-contact method of BE eradication and has been used both as primary and salvage therapy. We conducted a systematic review and meta-analysis to assess the safety and efficacy of LNSC. Methods We searched multiple databases from inception through December 2021 to identify studies on use of LNSC for Barrett's neoplasia. Pooled estimates were calculated using random-effects model and results were expressed in terms of pooled proportions with relevant 95 % confidence intervals (CIs) of complete eradication (CE) of dysplasia(D), high grade dysplasia (HGD) and IM. Results Fourteen studies with 707 patients were included in our final analysis. Overall pooled rates of CE-D, CE-HGD and CE-IM were 80.8 % (CI 77.4-83.8; I 2 62), 90.3 % (CI 85.2-93.7; I 2 33) and 55.8 % (CI 51.7-59.8; I 2 73) with follow up ranging from 4.25 months to 69.7 months. In patients with follow up beyond 24 months, the rates of CE-D and CE-IM were 83.6 % (CI 77.6-88.2; I 2 60) and 54.7 % (CI 47.6-61.6; I 2 81). Among LNSC naïve patients with prior history of endoscopic resection, the rates were 79.9 % (CI 73.3-85.2; I 2 50) and 67.1 % (CI 59.5-73.8; I 2 0). Pooled rate of therapeutic failures, defined as lack of response to LNSC therapy, was 23.6 % (CI 19.4-28.3; I 2 73). Post LNSC strictures and perforation pooled rates were 4 % and 0.8 %, respectively, which are similar to those previously reported for RFA. Conclusions Our analysis suggests that liquid nitrogen spray cryotherapy is an acceptable treatment for BE in both ablation naïve and experienced patients.

High Serum Squamous Cell Carcinoma Antigen Level Associated with Remission of Mild/Moderate Dysplasia of the Esophagus: A Nested Case-Control Study.

Background The esophageal epithelial dysplasia is the precancerous lesion. This study aimed to investigate the association between the serum squamous cell carcinoma antigen (SCCA) and the remission of esophageal squamous mild or moderate dysplasia. Methods We performed a nested case–control study. Patients with mild/moderate dysplasia of the esophageal squamous epithelium were enrolled in this study during the years of 2013–2015 and received a follow-up endoscopy during 2017–2018. With the comparison between baseline and follow-up diagnosis, the patients were divided into regression/stable and progression groups. A predictive model for the outcome of dysplasia was comprised of the variables of SCCA, age, sex, education level, and baseline dysplasia grade. A receiver operating characteristic (ROC) curve was used to estimate the diagnostic efficacy of the regression status of dysplasia under the predictive model. Results There were 146 patients enrolled in this study. 100 patients experienced a regression or stable status of dysplasia and 46 patients had a progressed status. Increased age, low education level, and moderate dysplasia were the risk factors of progression. With an 0.1 μg/L increase, SCCA was associated with a 0.90-fold risk (95% CI 0.81, 0.99) of progression. In the predictive model, the area under ROC curve was 0.78. The cut-off values of predictive probability of combined factors for progression, were 0.40 and 0.32 for males and females, respectively. Conclusions Increased serum SCCA concentration was associated with regressed severity of mild and moderate dysplasia of the esophageal mucosa. Further studies were warranted and SCCA concentration was a potential biomarker for the dysplasia prognosis.

S2400 Granular Cell Tumor: A Rare Disease of Esophagus

Introduction: Granular cell tumors (GCTs) are rare, often benign tumors of neurogenic origin. GCTs are usually found incidentally but can present with dysphagia in symptomatic patients. They are diagnosed via biopsy and histology following an esophagogastroduodenoscopy( EGD). We present a case of a 46-year-old female who presented with dysphagia, GERD, and weight loss diagnosed with esophageal granular cell tumor following an EGD. Case Description/Methods: A 46 years old female with a past medical history of H. pylori gastritis presents with complaints of dysphagia for 3 months and GERD associated with weight loss. EGD reviewed a single 4 mm nodule found in the lower third of the esophagus with the appearance of a lipoma (Figure). Biopsy of esophagus nodule resulted in esophageal squamous epithelium with focal granular cell tumor; Immunohistochemistry tumor cells are positive for S100, CD56, CD68, negative for calretinin and AE1/3 supporting rare esophageal granular cell tumor. EUS demonstrated a 4.7 mm x 2.6 mm intramural (subepithelial) lesion appeared to originate from within the superficial mucosa (Layer 1), Patient underwent repeat EGD with en-bloc cap and band-assisted endoscopic mucosal resection (EMR) followed by 3 hemoclips placed for hemostasis. The patient was discharged home on a proton pump inhibitor. The patient reports clinical improvement on follow-up after 4 weeks. Discussion: GCTs are thought to originate from Schwann cells; the prevalence of GCTs is challenging to determine as there have been roughly 300 documented cases of GCTs discussed in the literature. Conversely, patients with larger diameter lesions typically present with indolent dysphagia and other less common symptoms such as gastro-esophageal reflux disease, dyspepsia, chest pain, cough, or nausea. GCTs on EGD appear sessile, yellow-white in color, and firm, with a negative pillow sign. After performing EGD, an endoscopic ultrasound (EUS) is diagnostically utilized to ascertain tumor invasion and provide a biopsy through fine needle aspiration (FNA). Histopathological diagnosis, including immunohistological staining, is essential for confirmation of GCT. Immunohistological staining is positive for S-100, PAS (Periodic Acid Schiff), neuron-specific enolase, nestin, and vimentin. Patients whose neoplasms are less than 1 cm in diameter are treated conservatively with endoscopic follow-up, with tumors greater than 1 cm in diameter, or having clinical symptoms that need endoscopic or surgical resection.Figure 1.: A: EUS showing 4.7 mm x 2.6 mm hypoechoic intramural (subepithelial) lesion was found in the lower third of the esophagus. It appeared to originate from within the superficial mucosa (Layer 1). A tissue diagnosis was obtained prior to this exam. B: EGD significant for sessile, yellow white in color, and firm mass followed by EMR. C: Histopathology with H&E showing clusters of polygonal cells with granular eosinophilic cytoplasm (top) and S100 stain (below).

S452 Safety and Efficacy of Liquid Nitrogen Spray Cryotherapy in Barrett’s Neoplasia: A Comprehensive Review and Meta-Analysis

Introduction: Barrett’s esophagus (BE) is a precursor condition to esophageal adenocarcinoma (EAC), resulting in transformation of the squamous epithelium of distal esophagus to columnar-lined epithelium with intestinal metaplasia (IM). Liquid nitrogen spray cryotherapy (LNSC) is a non-contact method of BE eradication and has been used both as primary and salvage therapy. We conducted a systematic review and meta-analysis to assess the safety and efficacy of LNSC. Methods: We searched multiple databases from inception through December 2021 to identify studies on use of LNSC for Barrett’s neoplasia. Pooled estimates were calculated using random-effects model and results were expressed in terms of pooled proportions with relevant 95% confidence intervals (CIs) of complete eradication (CE) of dysplasia(D), high grade dysplasia (HGD) and IM. We also assessed pooled rates of therapeutic failures as well as post procedure complications including strictures, pain and perforation events. Results: Fourteen studies with 666 patients were included in our final analysis. (Figure) Overall pooled rates of CE-D, CE-HGD and CE-IM were 80.8% (CI 77.4-83.8; I2 62), 87% (CI 82-90.8; I2 60) and 55.8% (CI 51.7-59.8; I2 73) with follow up ranging from 4.25 months to 69.7 months. Among LNSC naïve patients with prior history of endoscopic resection, the rates were 83% (CI 73.6-89.5; I2 46), 76% (CI 65.1-84.3; I2 38) and 71.8% (CI 60.9-80.7; I2 0). In patients with follow up beyond 24 months, the rates were 78.8% (CI 72.4-84.1; I2 84), 85.2% (CI 73.1-92.4; I2 1) and 54.7% (CI 47.6-61.6; I2 81). Pooled rate of therapeutic failures, defined as lack of response to LNSC therapy, was 22.1% (CI 18.3-36.6; I2 74). Post LNSC strictures and perforation pooled rates were 4% and 0.8%, respectively, which are lower than previously reported for RFA. Pooled rate of post procedure abdominal and/or chest pain was 13.6% (CI 11.3-16.3; I2 67). (Table) Conclusion: Our analysis suggests that liquid nitrogen spray cryotherapy is a durable and safe therapeutic modality for BE, in both ablation naïve and experienced patients.Figure 1.: Study analysis Table 1. - Study Outcomes Recurrence Adverse Events Failure F/u Time CE-D (CE-HGD, CE-LGD) CE-IM Dysplasia IM Pain Stricture Perforation Dysphagia Dumot 2009 CE-D 15/ 31 CE-IM 6/ 31 16/25 (LGD 6, HGD/IMCA/ACA, esophagectomy, or death 10) -- 10/31 Mild pain 7/31, severe pain 3/31 3/31 1/31 -- 9/31 12m (6-24) Greenwald 2010 CE-D 22/24 - 15/17 (HGD), 4/4 (ImCA), 3/3 (Barrett’s Carcinoma); CE-HGD - 23/24 CE-IM 13/24 - 9/17 (HGD), 3/4 (ImCA), 1/3 (Barrett’s Carcinoma) -- -- CP 57/323, Abdominal pain 14/323 (procedures) 3/77 1/77 43/323 (procedures) -- 9.3 (3,13) - 13.8 (10,18) Shaheen 2010 CE-D 52/60, CE-HGD 58/60 CE-IM 34/60 -- -- 2/98 3/98 0/98 - -- 10.5m (8.3) [Mean] Halsey 2011 CE-D and CE-HGD 25/36 -- 6/36 5/36 -- -- -- -- 3/36 24m Sengupta 2015 CE-D 12/ 16 CE-IM 5/ 16 after CE-D 0/12 -- -- 3/21 1/21 3/21 -- 7.5m (2-25m) Ghorbani 2016 CE-D 67/80 [HGD: CE-HGD 52/57, CE-D 46/57; LGD: CE-D 21/23] CE-IM 51/80 [HGD: CE-IM 37/57; LGD: CE-IM 14/23] -- -- 28/96. Abdo pain 6/96, CP 22/96 1/96 0/96 11/96 -- 21 m (12–24 m) Suchniak - Mussari 2017 CE-D 17/20 (CE-HGD 14/15, CE-LGD 3/5); CE-ImCA+D - 6/8 CE-IM 16/33 -- -- -- 5/45 0/45 -- 1/15 (with HGD), 2/8 (with ImCA) 27.6m (13.2) [Mean/SD] Ramay 2017 CE-D: 27/36 (CE-HGD 32/39) CE-IM 17/26 after CE-D: 7/39 (HGD) -- -- -- -- -- 13/40 (LGD 3, IM 10) 69.7 (13.8) [53.8-106.0] {Mean/SD} Trindade 2017 CE-D 13/18 (CE-LGD 3/4, CE-HGD 5/7) CE-IM 9/ 18 after CE-D 0/18 after CE-IM 0/18 0/18 0/18 -- -- -- 4.25m (3–11) [Median] Trindade 2018 CE-D 22/27 CE-IM 19/27 -- after CE-IM 9/ 27 0 0 0 0 8/27 (2 LGD, 3 IM, 3 cancer) 2 y [1-5.5] [Median] Thota 2018 CE-D 63/81 CE-IM 33/81 -- after CE-IM 9/63 -- -- -- -- 10/81 31.8m [12.6, 50.7] {Median} Spiceland 2019 CE-D 38/46 CE-IM 21/46 -- -- -- 3/46 -- -- 8/46 (1 cancer, 2 lost f/u, 5 undergoing rx) NR Kaul 2020 CE-D -51/52 CE-IM - 39/52; LGD 6/7; HGD 13/17; T1a 13/17; inv EAC 7/11 4/34 after CE-IM 3/34 0/57 0/57 1/57 0/57 20/39 4.8 yr Fasullo 2021 CE-D 44/62 CE-IM 41/62 6/44 after CE-IM 6/41 0/62 0/62 0/62 -- 11/62 NR

S2353 Esophageal Hyperkeratosis Complicated by Recurrent Dysphagia

Introduction: Esophageal hyperkeratosis (EH) is a rare condition characterized by keratinization of the esophageal epithelium. The pathophysiology is poorly understood, and risk factors remain unclear. We present a case of severe long segment EH leading to severe esophageal stenosis. Case Description/Methods: A 70-year-old male with a long history of gastroesophageal reflux disorder (GERD) presented to the ED with acute food impaction. Prior to his presentation, he reported worsening dysphagia and ongoing heartburn for several months. He had been lost to follow-up and was not on any antacids at the time of presentation. Emergent upper endoscopy (EGD) was performed to remove the food impaction. Additional findings on EGD showed esophagitis, esophageal stenosis of 13cm in length (7mm in diameter), and white plaques. Esophageal biopsies were negative for eosinophils but confirmed candida and were suggestive of esophageal hyperkeratosis. Dilation was deferred until the esophagitis was treated with PPI and itraconazole. Repeat EGD one month later showed ongoing stenosis, healed esophagitis, as well as significantly thickened esophageal mucosa extending 13 cm proximally from the GE junction (Figure A, B). Balloon dilation of the stenotic lesion was performed. Repeat biopsies confirmed esophageal hyperkeratotic squamous epithelium with parakeratosis and no dysplasia (Figure C). Due to the severity of his stenosis, he subsequently underwent multiple EGDs with balloon dilations over the next several months to obtain a maximum esophageal lumen diameter of 19mm. Dilation of his esophagus was particularly difficult due to the length of hyperkeratosis, degree of stenosis, and loss of elasticity of his esophageal mucosa. Tearing of the tissue and bleeding were common even with small interval increases during each dilation session. In addition, healing of the tissue between sessions often led to a restricturing of his esophagus. He is currently symptom-free on pantoprazole 40mg twice a day, a soft diet, and undergoing surveillance endoscopies every 6 months with balloon dilations. Discussion: Esophageal hyperkeratosis is a rare disorder with thickening of the stratum corneum and occurs in 2% of esophageal biopsies. While the prevalence and pathophysiology are unclear, it is thought to be associated with GERD, vitamin A deficiency, zinc deficiency, and a low fiber/low residue diet. There is also some evidence to suggest it may be associated with an increased risk of squamous cell carcinoma.Figure 1.: 1: a and b) EGD of middle and lower esophagus showing irregular mucosa with furrowing and white exudate, pre-dilation. 1c) Magnification 400x. Histopathology demonstrates esophageal squamous mucosa with dyskeratosis and extensive hyperkeratosis (black arrow).

420. IDENTIFICATION AND CLASSIFICATION OF TUMOR CELLS IN PATIENTS WITH BARRETT’S CARCINOMA BY HYPERSPECTRAL IMAGING (HSI)

Abstract Hyperspectral imaging (HSI), as recently applied in medicine, is a novel technology combining imaging with spectroscopy. It might be used to identify, classify and discriminate malignant and non-malignant cells of histopathologic specimens. HSI allows the determination of a spectrum between the visual and near-infrared light (500-1000 nm). After surgical resection, specimens (n = 96) of Barrett’s cancer were fixed in 4% formaldehyde and slices were conducted (3 μm), which were stained with hematoxylin and eosin (HE). Differences in the absorbance of squamous epithelium and esophageal adenocarcinoma (EAC) cells were determined at eosin’s and hematoxylin’s maximal absorption of 530 nm and 590 nm. A classification algorithm, which combines a multilayer perceptron (MLP) with a Gaussian filter and data reduction based on principal component analysis (PCA) was used for discrimination. For the first time in literature, we were able to analyze esophageal adenocarcinoma, tumor stroma and squamous epithelium cells by HSI. For both, the squamous epithelium and the esophageal adenocarcinoma cells, the intragroup variances were quite low. A set of 20 specimens was used in a testing phase, which showed MLP by using reflectance data to provide the best results. Leave-one-patient-out-cross validation for all 96 specimens showed an accuracy of 77% with a sensitivity of 87% and a specificity of 72% for EAC determination and visualization. Squamous epithelium and EAC cells show specific spectral alterations due to their HE-staining, when measured by HSI. However, the training algorithms need further validation to foster a semi-automatic decision-making process in histopathological tumor cell identification.

428. CHARACTERIZATION OF CD82/KAI1 IN ESOPHAGEAL ADENOCARCINOMA

Abstract The incidence of esophageal adencocarcinoma (EAC) has increased in last decades. Regardless to improved perioperative chemotherapies and surgical approaches, the prognosis for EAC in advanced stages is poor. CD82 is a tumor suppressor gene, and its downregulation correlates with invasive growth, metastases formation and advanced clinical stages. CD82 might influence the Wnt/ß-catenin signaling pathway. We used an in vitro Barrett’s esophagus cell culture model, represented by esophageal squamous cell epithelium (EPC1, EPC2), metaplasia (CP-A), dysplasia (CP-B) and esophageal adenocarcinoma (OE33, OE19, SKGT4, FLO1). Antisense LNA Oligonucleotides (GapmeRs, Qiagen) were used to achieve a transient knockdown of CD82 in OE33 and OE19 cells. mRNA and proteins were analyzed by q-RT-PCR and western blot analyses, respectively. A fluorescence antibody specific to CD82 was used for flow cytometry analysis. Proliferation assay, colony formation assay and boyden chamber assay were performed after CD82-knockdown to further analyze cell proliferation, cell growth and survival, colony forming and cell migration. CD82 was expressed in all investigated cell lines by flow cytometry with a significant overexpression in OE19 and FLO1 cells. A stable downregulation of CD82 was achieved in OE33 and OE19 cells. Proliferation assays showed a significantly higher proliferation rate at 72 hours after CD82-knockdown in OE33 cells. A significantly higher mRNA expression of vimentin was observed after CD82-knockdown in OE33. Colony formation assay showed a significantly higher colony count after CD82-knockdown in both cell lines. Cell migration was increased after CD82-downregulated in OE33 and OE19 cells. CD82 might influence the expression of mesenchymal marker proteins, cell migration abilities and the process of colony forming and thus enhance the metastatic potential of OE33 and OE19 cells. Low expression of CD82 could alter metastasis and cancer progression but further molecular characterizations are needed to elucidate the impact on carcinogenesis of EAC.

Identifying LATS2 as a prognostic biomarker relevant to immune infiltrates in human esophageal squamous cell carcinoma.

According to the TIMER database, large tumor suppressor 2 (LATS2) is differentially expressed in various tumors. However, the correlation between LATS2 and esophageal squamous cell carcinoma (ESCC) and the association between LATS2 and immune infiltration in ESCC remain unclear. Our synthetic research on LATS2 in ESCC revealed that the expression was low in esophageal squamous epithelium tissues, revealing the pernicious and adverse prognosis of ESCC. The Kaplan–Meier survival investigation pointed out that low LATS2 expression would result in an adverse prognosis. Biological investigation indicated that LATS2 was engaged in cell migration, adhesion, and junction. To further explore the relationship between LATS2 and tumor immunity, we utilized CIBERSORT to assess immune infiltration. The findings revealed that specimens with lower LATS2 expression showed higher immune infiltration, including T-cell follicular helper cells, M0 macrophages, M1 macrophages, and myeloid dendritic cell resting. An association investigation indicated that LATS2 was negatively relevant to immune checkpoints that restrain operative antitumor immune reactions. We also conducted immunohistochemical staining to explore the link between LATS2 expression and immunophenotype. The indicated association between low LATS2 expression and an immunophenotype is conducive to our understanding of ESCC mini-environments and might offer new indications for enhancing new therapeutic targets.