Gallbladder Carcinoma Patients Research Articles

ARRB1 Drives Gallbladder Cancer Progression by Facilitating TAK1/MAPK Signaling Activation.

Gallbladder carcinoma (GBC) is the most common malignancy of the biliary tract, with a dismal 5-year survival of 5%. Recently, ARRB1, as a molecular scaffold, has been proposed to participate in the progression of multiple malignancies. However, the effect and regulatory mechanisms of ARRB1 in GBC have not been investigated. Our study aimed to explore the biological functional status and the possible molecular mechanisms of ARRB1 with respect to GBC progression. The survey showed that human GBC tissues exhibited increased levels of ARRB1 compared with normal tissues, and the high expression of ARRB1 was associated with poor prognosis of GBC patients. A series of in vitro and in vivo functional experiments based on knockdown of ARRB1 uncovered that ARRB1 enhanced GBC cell proliferation, migration, and invasion. Furthermore, we reported that TAK1, a component of the TNF /MAPK pathway, is a vital downstream effector of ARRB1. In addition, siTAK1 could abolish the functional changes between ARRB1 overexpression GBC cells and control ones. Our data revealed that ARRB1 facilitated the carcinogenesis and development of GBC through TNF/TAK1/MAPK axis, suggesting that ARRB1 may be a promising biomarker and treatment target for GBC patients.

Prediction of Malignancy Using Skin Autofluorescence in Patients with Gallbladder Carcinoma

Introduction: Increased accumulation of advanced glycation end products (AGEs) may correlate with progression of numerous diseases, including some cancers. Skin autofluorescence (SAF) measurement in a non-invasive test that quantifies tissue AGEs. The aim of this study was to assess utility of SAF in prediction of malignancy in patients with gallbladder carcinoma (GBC). Methods: This prospective study comprised 29 patients (7 men and 22 women) with suspected or confirmed GBC who underwent surgery in the Department of General, Transplant and Liver Surgery of the Medical University of Warsaw between September 2018 and November 2019. SAF was based on 3 separate preoperative measurements on the anterior side of the forearm and expressed in arbitrary units (AU). Data on patients' weight, height, BMI, and diabetes were collected preoperatively. Results: Gallbladder carcinoma was diagnosed in 22 out of 29 patients (75.86%). Median SAF was 2.2 AU (IQR = 1.925 − 2.675) and 2.2 AU (IQR = 1.8 − 2.5) in GBC patients and individuals without cancer, respectively. There was no difference in SAF between patients with GBC and individuals without cancer (p=0.700). The lack of significant difference was unmodified by patient sex, presence of diabetes or BMI>25 (p=0.662, p=0.643, and p=0.629, respectively). Conclusions: Although SAF may have predictive value in some cancers, this study does not provide evidence for increased accumulation of AGEs in patients with GBC.

MiR-1231 enhances docetaxel sensitivity to gallbladder carcinoma cells by downregulating FOXC2.

To illustrate the role of microRNA-1231 (miR-1231) in regulating malignant proliferative potential and DTX sensitivity to gallbladder carcinoma (GBC) by regulating FOXC2 level. Expression levels of miR-1231 in GBC tissues and paracancerous ones were detected. The relationship between miR-1231 level and clinical parameters of GBC patients was analyzed. After overexpression of miR-1231, changes in proliferative and apoptotic potentials in GBC-SD and NOZ cells were examined by Cell Counting Kit-8 (CCK-8), colony formation assay and flow cytometry, respectively. Regulatory effects of miR-1231 on its downstream gene FOXC2 were determined by Luciferase assay. Finally, the role of miR-1231 in regulating DTX sensitivity to GBC cells was assessed. MiR-1231 was downregulated in GBC tissues compared to paracancerous ones. GBC patients expressing lower level of miR-1231 had worse tumor staging and larger tumor size. Overexpression of miR-1231 attenuated proliferative potential, and induced apoptosis in GBC cells. FOXC2 was upregulated in GBC and negatively linked to miR-1231. Luciferase activity confirmed that FOXC2 was the target gene binding miR-1231. DTX treatment dose-dependently suppressed viability in GBC cells and overexpression of miR-1231 could enhance DTX sensitivity in GBC. Notably, overexpression of FOXC2 abolished regulatory effects of overexpressed miR-1231 on proliferative and apoptotic potentials in GBC cells. MiR-1231 is downregulated in GBC species. Its level is closely linked to tumor staging and tumor size in GBC patients. By downregulating FOXC2, miR-1231 enhances DTX sensitivity to GBC cells and thus alleviates the malignant development of GBC.

Immunoproteomics approach revealed elevated autoantibody levels against ANXA1 in early stage gallbladder carcinoma

BackgroundEarly diagnosis is important for the timely treatment of gallbladder carcinoma (GBC) patients and may lead to increased survival outcomes. Here, we have applied serological proteome analysis (SERPA), an immunoproteomics approach, for the detection of ‘tumor-associated antigens (TAAs) that elicit humoral response’ in early stage GBC patients.MethodsTotal protein from pooled tumor tissue of GBC patients (n = 7) was resolved by two-dimensional gel electrophoresis (2-DE) followed by immunoblotting using pooled blood plasma from healthy volunteers (n = 11) or gallstone disease (GSD) cases (n = 11) or early stage GBC (Stage I and II) (n = 5) or GBC stage IIIA (n = 9). 2-D gel and immunoblot images were acquired and analyzed using PDQuest software to identify immunoreactive spots in GBC cases in comparison to controls. Proteins from immunoreactive spots were identified by liquid chromatography- tandem mass spectrometric analysis (LC-MS/MS). Autoantibody levels for two of the functionally relevant proteins were investigated in individual plasma samples (52 cases and 89 controls) by dot blot assay using recombinant proteins.ResultsImage analysis using PDQuest software identified 25 protein spots with significantly high or specific immunoreactivity in GBC cases. Mass spectrometric analysis of 8 corresponding protein spots showing intense immunoreactivity (based on densitometric analysis) in early stage GBC or GBC stage IIIA cases led to the identification of 27 proteins. Some of the identified proteins include ANXA1, HSPD1, CA1, CA2, ALDOA and CTSD. Among the two proteins, namely ANXA1 and HSPD1 verified using a cohort of samples, significantly elevated autoantibody levels against ANXA1 were observed in early stage GBC cases in comparison to healthy volunteers or GSD cases (unpaired t-test, p < 0.05). Receiver operating characteristic (ROC) curve analysis for ANXA1 showed an Area under the Curve (AUC) of 0.69, with 41.7% sensitivity against a specificity of 89.9% for early stage GBC. IHC analysis for ANXA1 protein showed ‘high’ expression levels in 72% of GBC cases whereas all the controls showed ‘low’ expression levels.ConclusionsThe study suggests that the ANXA1 autoantibody levels against ANXA1 may be potentially employed for early stage detection of GBC patients. Other proteins could also be explored and verified in a large cohort of clinical samples.

Modification of the 8th American Joint Committee on Cancer staging system for gallbladder carcinoma to improve prognostic precision

BackgroundThe 8th edition of the American Joint Committee on Cancer (AJCC) staging system for gallbladder carcinoma (GBC) came into force since 2018. However, the prognostic precision of this staging system has not been properly assessed. This study aimed to evaluate the latest staging system and suggest modifications to improve its prognostic precision.MethodsData of patients with GBC was included from the Surveillance, Epidemiology and End Results (SEER) database (2004–2015) and multicenter database (2010–2017). Baseline clinicopathologic characteristics were recorded including age, sex, race, grade, T category, N category, M category and stage. The Kaplan-Meier method was used to plot survival functions. The prediction power of the AJCC 8th edition and its modified version were evaluated using the concordance index (C-index).ResultsA total of 2779 GBC patients were included in the SEER database and 591 were collected from multicenter database. While no significant difference in survival of patients was observed between stages IVA and IVB using the 8th AJCC staging system (p > 0.05), the prognosis of stage IIIA showed a slightly better outcome than stage IIIB (p = 0.046) in the SEER database. In the multicenter database, there was no significant difference between stage IIIA and stage IIIB (p > 0.05). Similarly, no significant difference in the survival of patients between stages IIIA and IIIB was observed when M0 patients with at least 6 lymph nodes (LNs) were analyzed (p > 0.05) for both SEER and multicenter database. On the other hand, a modified staging system was able to stratify patients from stage IIIA, stage IIIB and stage IV (p < 0.001). For the SEER database, the C-indexes of 8th AJCC staging system and that of its modified version were 0.709 and 0.742, respectively. For the multicenter database, the C-index of 8th AJCC staging system and that of our modified version were 0.635 and 0.679, respectively.ConclusionsThe modified 8th staging system proposed in this study can improve the prognostic precision of the 8th AJCC staging system for GBC. We therefore suggest including these modifications in the next update of AJCC staging system for GBC.

Influence of adjuvant therapy on pattern of failure and survival in curatively resected gallbladder carcinoma.

The study was done to evaluate the role of adjuvant therapy in curatively resected Stage II and III gallbladder carcinoma (GBC). This was a retrospective analysis of patients of GBC registered between 2008 and 2017 in outpatient department of a tertiary cancer hospital in India. Patients who had any of the following adjuvant treatment after radical surgery: (a) external beam radiotherapy (RT) alone, (b) chemotherapy (CT) alone, and (c) RT with CT (CRT) were considered for the study. A total of fifty patients could meet the selection criteria. It was seen that seven patients were treated with RT, 20 with CT, and 23 with CRT. Median follow-up for patients who were alive was 26.7 months. Nineteen patients had locoregional failure while eight had distant failure. Patients treated with CRT had a significantly better mean overall survival compared to those treated with RT or CT (44.0 months, 12.5 months, and 15.1 months, respectively; P = 0.003). Similarly, mean disease-free survival was superior in CRT arm compared to RT and CT arms (43.6 months, 9.6 months, and 12.4 months, respectively; P = 0.002). Adjuvant CRT had better survival outcome compared to patients treated with either RT or CT with Stage II and III disease after curative cholecystectomy.

The Impacts of Systemic Immune-Inflammation Index on Clinical Outcomes in Gallbladder Carcinoma.

Background: Systemic immune-inflammation index (SII) is considered to be a prognostic marker in several cancers. However, the prognostic value of baseline pre-operative SII in gallbladder carcinoma (GBC) has not been evaluated. This study aimed to determine the prognostic significance of SII and generate a predictive nomogram.Methods: We retrospectively studied 142 GBC patients who underwent surgical resection at the Peking Union Medical College Hospital between 2003 and 2017. SII, neutrophil-to-lymphocyte ratio (NLR), and lymphocyte-to-monocyte ratio (LMR) were evaluated for their prognostic values. A multivariate Cox proportional hazards model was used for the recognition of significant factors. Then, the cohort was randomly divided into the training and the validation set. A nomogram was constructed using SII and other selected indicators in the training set. C-index, calibration plots, and decision curve analysis were performed to assess the nomogram's clinical utility in both the training and the validation set.Results: The predictive accuracy of SII (Harrell's concordance index [C-index]: 0.624), NLR (C-index: 0.626), and LMR (C-index: 0.622) was evaluated. The multivariate Cox model showed that SII was a superior independent predictor than NLR and LMR. SII level (≥600) (hazard ratio [HR]: 1.694, 95% confidence interval [CI]: 1.069–2.684, p = 0.024), carbohydrate antigen (CA) 19-9 level (≥37 U/ml) (HR: 2.407, 95% CI: 1.472–3.933, p < 0.001), and TNM stage (p = 0.026) were selected to construct a nomogram for predicting overall survival (OS). The predictive ability of this model was assessed by C-index (0.755 in the training set, 0.754 in the validation set). Good performance was demonstrated by the calibration plot. A high net benefit was proven by decision curve analysis (DCA).Conclusion: SII is an independent prognostic indicator in GBC patients after surgical resection, and the nomogram based on it is a useful tool for predicting OS.

Long non-coding RNA Linc00261 as a novel potential diagnostic and prognostic biomarker for gallbladder cancer.

BackgroundLinc00261 is a lncRNA that plays key roles in tumor suppression. While gallbladder carcinoma (GBC) is one of the most common cancer of the bile duct. However, the study about Linc00261’s correlation with the clinicopathological characteristics and postoperative outcomes of the GBC patients is few. Therefore, we want to explore Linc00261 in GBC and assess its potential of clinical diagnosis.MethodsQuantitative real-time PCR (qRT-PCR) was used to detect the expression of Linc00261 in specimens of GBC and adjacent tissues as well as cell lines. Chi-square test has been used to research the correlation of the Linc00261 expression in GBC with the clinicopathological features. The Cox model was used to assess the value of Linc00261 in predicting the prognosis of GBC patients. ROC curve analysis was used to test the specificity and sensitivity of diagnostic method of serum Linc00261 expression.ResultsThe expression level of Linc00261 in GBC was significantly lower than normal tissues’ and it was also up-regulated after surgery. The Linc00261 expression was significantly correlated with large tumor size (P<0.0001), late TNM stage (P=0.008), negative liver metastasis (P=0.027) and well differentiated phenotype (P=0.017). The patients with lower Linc00261 expression had significantly worse outcomes in terms of overall survival (P=0.0188) and progression-free survival (P=0.0029), and the low expression of Linc00261 was identified as an independent risk factor affecting postoperative survival rate of the patients (P<0.01). The expression of Linc00261 in serum was down-regulated of GBC patients and increased in the patients after operation. Linc00261 expressed in serum was also positively associated with its expression in GBC tissue of patients (P<0.0001). The GBC diagnosis efficacy of using the serum Linc00261 level to identify the GBC has high specificity and sensitivity (AUC 0.805).ConclusionsLinc00261 could be identified a novel gene associated with GBC development and progression. It also may serve as a new diagnostic and prognostic biomarker for patients with GBC.

Loss of desmoglein-2 promotes gallbladder carcinoma progression and resistance to EGFR-targeted therapy through Src kinase activation

Gallbladder carcinoma (GBC) exhibits poor prognosis due to local recurrence, metastasis, and resistance to targeted therapies. Using clinicopathological analyses of GBC patients along with molecular in vitro and tumor in vivo analysis of GBC cells, we showed that reduction of Dsg2 expression was highly associated with higher T stage, more perineural, and lymphatic invasion. Dsg2-depleted GBC cells exhibited significantly enhanced proliferation, migration, and invasiveness in vitro and tumor growth and metastasis in vivo through Src-mediated signaling activation. Interestingly, Dsg2 binding inhibited Src activation, whereas its loss activated cSrc-mediated EGFR plasma membrane clearance and cytoplasmic localization, which was associated with acquired EGFR-targeted therapy resistance and decreased overall survival. Inhibition of Src activity by dasatinib enhanced therapeutic response to anti-EGFR therapy. Dsg2 status can help stratify predicted patient response to anti-EGFR therapy and Src inhibition could be a promising strategy to improve the clinical efficacy of EGFR-targeted therapy.

γ-glutamyl transferase-to-platelet ratio based nomogram predicting overall survival of gallbladder carcinoma.

BACKGROUNDGallbladder carcinoma (GBC) carries a poor prognosis and requires a prediction method. Gamma-glutamyl transferase–to–platelet ratio (GPR) is a recently reported cancer prognostic factor. Although the mechanism for the relationship between GPR and poor cancer prognosis remains unclear, studies have demonstrated the clinical effect of both gamma-glutamyl transferase and platelet count on GBC and related gallbladder diseases.AIMTo assess the prognostic value of GPR and to design a prognostic nomogram for GBC.METHODSThe analysis involved 130 GBC patients who underwent surgery at Peking Union Medical College Hospital from December 2003 to April 2017. The patients were stratified into a high- or low-GPR group. The predictive ability of GPR was evaluated by Kaplan–Meier analysis and a Cox regression model. We developed a nomogram based on GPR, which we verified using calibration curves. The nomogram and other prognosis prediction models were compared using time-dependent receiver operating characteristic curves and the concordance index.RESULTSPatients in the high-GPR group had a higher risk of jaundice, were older, and had higher carbohydrate antigen 19-9 levels and worse postoperative outcomes. Univariate analysis revealed that GPR, age, body mass index, tumor–node–metastasis (TNM) stage, jaundice, cancer cell differentiation degree, and carcinoembryonic antigen and carbohydrate antigen 19-9 levels were related to overall survival (OS). Multivariate analysis confirmed that GPR, body mass index, age, and TNM stage were independent predictors of poor OS. Calibration curves were highly consistent with actual observations. Comparisons of time-dependent receiver operating characteristic curves and the concordance index showed advantages for the nomogram over TNM staging.CONCLUSIONGPR is an independent predictor of GBC prognosis, and nomogram-integrated GPR is a promising predictive model for OS in GBC.

Effect of preoperative jaundice on long-term prognosis of gallbladder carcinoma with radical resection

PurposesThis study was designed to evaluate the effect of preoperative jaundice on long-term prognosis of gallbladder carcinoma (GBC) after radical resection (R0).MethodsA total of 267 GBC patients who underwent R0 resection from January 2004 to December 2014 were enrolled, including 54 patients with preoperative jaundice and 213 patients without jaundice. The clinicopathological parameters between the two groups were compared, and the correlation between preoperative jaundice and the long-term prognosis was furtherly analyzed.ResultsUnilateral and multivariate analyses of 267 GBC patients showed that the depth of tumor invasion (pT stage), lymphatic metastasis, and hepatic invasion were independent prognostic factors. The univariate and multivariate analysis of 54 GBC patients with preoperative jaundice showed that only pT stage was an independent factor for prognosis. Furthermore, the intraoperative blood transfusion and pT stage were significant different between long-term survival (survive for more than 3 years) and those who died within 3 years (P < 0.05).ConclusionPreoperative jaundice was not the independent factor resulting in the poor long-term prognosis of gallbladder carcinoma after R0 resection. The pT stage was the only long-term prognostic factor in all GBC patients regardless of preoperative jaundice.

Raised CA19\u20139 and CEA have prognostic relevance in gallbladder carcinoma

BackgroundRole of tumor markers in gall bladder carcinoma (GBC) is not well established. We evaluated the prognostic value of carbohydrate antigen 19–9 (CA19–9) and carcinoma embryonic antigen (CEA) in patients with GBC.MethodsOf the 225 patients of GBC enrolled,176 patients were included in the study (excluded 49 patients with jaundice). Patients were divided into 3 groups; resectable n = 92, unresectable n = 17, metastatic n = 67. The clinico-pathological characteristics, tumor markers and survival data were analysed. The cutoff values of CA19–9 & CEA for predicting metastases were computed using receiver operating characteristic curve. Kaplan Meir survival and Cox regression analysis were done for factors predicting survival and recurrence.ResultsThe median value of Ca19–9 was significantly higher in metastatic group [resectable: 21.3, unresectable: 53.9 and metastatic: 79; p < 0.001] but not for CEA [3.5, 7.8 and 5 ng/ml (p = 0.20)]. A cutoff value of 72 IU/ml for CA19–9, 5 ng/ml for CEA had a sensitivity and specificity of 52 and 80%, 51 and 72% respectively for detection of metastatic disease. Median, 3-year & 5-year survival were significantly lower in patients with CEA > 4 (p = 0.041), Ca19.9 > 37 (p = 0.019), T3/T4 (p = 0.001), node positive (p = 0.001) and presence of perineural invasion (p = 0.001). However, on multivariate analysis, only Ca19.9 > 37 predicted recurrence (p = 0.002, HR 5.8).ConclusionsRaised CA19.9 and CEA predict metastatic disease in patients with GBC without jaundice with a high specificity and may help in prognostication of the patient. CA19–9 was better than CEA in prediction of tumor burden and in predicting recurrence.

Circular RNA circ-MTO1 serves as a novel potential diagnostic and prognostic biomarker for gallbladder cancer.

Primary gallbladder carcinoma (GBC) is one of the most common biliary malignancies in the gastrointestinal tract. In this work, we examined the roles of circular-mitochondrial translation optimization 1 (circ-MTO1) in GBC tissues and patient plasma. Circ-MTO1 expression in GBC tissues and patient plasma was evaluated by quantitative Real Time-PCR (qRT-PCR). The relationships between circ-MTO1 expression and the pathological characteristics of GBC were analyzed. Kaplan-Meier survival curve was applied to calculate overall survival (OS) and progression-free survival (PFS) in GBC patients with different circ-MTO1 expression. The univariate COX regression curve analysis method was employed to analyze the potential relationships between high circ-MTO1 expression and OS and PFS. At last, we assessed the diagnostic value of the circ-MTO1 level in GBC patient plasma. Circ-MTO1 expression was significantly upregulated in tumor tissues and plasma in GBC patients. In addition, circ-MTO1 expression was associated with clinical-pathological characteristics in GBC. High circ-MTO1 expression served as an independent prognostic factor for poor OS and PFS in GBC patients. Moreover, upregulated plasma circ-MTO1 level was significantly associated with tumor development. Circ-MTO1 is a potential early diagnostic and prognostic biomarker for patients with gallbladder cancer. Thus, our present work might provide a new understanding of the diagnosis and treatment of GBC.

Prognostic value of preoperative lymphocyte-to-monocyte ratio in gallbladder carcinoma patients and the establishment of a prognostic nomogram.

The purpose of this study was to investigate the potential prognostic value of preoperative lymphocyte-to-monocyte ratio (LMR) and establishment of a prognostic nomogram in post surgical patients with gallbladder carcinoma (GBC).Receiver operating characteristic curve analysis was performed to determine the optimal cut-off value of LMR. The correlation between preoperative LMR and overall survival (OS) was analyzed using univariate and multivariate Cox regression analyses. A relevant prognostic nomogram was established.Three hundred fifteen GBC patients were retrospectively enrolled. Based on receiver operating characteristic curve analysis, the optimal cutoff value of LMR was 2.685. Patients were categorized into high-LMR group (n = 143) or low-LMR group (n = 172). Low-LMR value was significantly associated with elderly age, advanced tumor, and the performance of a palliative cholecystectomy. The results of the univariate and multivariate analyses eliminated the degree of tumor differentiation, tumor-node-metastasis stages, surgery types, and LMR as independent predictors of OS. Based on those independent predictors, a predictive nomogram for OS was generated with an accuracy of 0.848.Based on our findings, the predictive nomogram should be included in the routine assessment of GBC patients.

Survival prediction for gallbladder carcinoma after curative resection: Comparison of nomogram and Bayesian network models

BackgroundIn this study, we developed a nomogram and a Bayesian network (BN) model for prediction of survival in gallbladder carcinoma (GBC) patients following surgery and compared the performance of the two models. MethodsSurvival prediction models were established and validated using data from 698 patients with GBC who underwent curative-intent resection between 2008 and 2017 at one of six Chinese tertiary hospitals. Model construction and internal validation were performed using data from 381 patients at one hepatobiliary center, and external validation was then performed using data from 317 patients at the other five centers. A BN model and a nomogram model were constructed based on the independent prognostic variables. Performance of the BN and nomogram models was compared based on area under receiver operating characteristic curves (AUC), model accuracy, and a confusion matrix. ResultsIndependent prognostic variables included age, pathological grade, liver infiltration, T stage, N stage, and margin. In internal validation, AUC was 84.14% and 78.22% for the BN and nomogram, respectively, and model accuracy was 75.65% and 72.17%, respectively. In external validation, AUC was 76.46% and 70.19% for the BN and nomogram, respectively, with model accuracy of 66.88% and 60.25%, respectively. Based on the confusion matrix, the nomogram had a higher true positive rate but a substantially lower true negative rate compared to the BN. ConclusionA BN model was more accurate than a Cox regression-based nomogram for prediction of survival in GBC patients undergoing curative-intent resection.

Abstract A10: Gallbladder cancer disparities in New Mexico: Examining the role of environmental heavy metal exposures as a driver of gallbladder epithelial signaling dysfunction

Abstract Introduction: Gallbladder carcinoma (GBC) is a rare, yet deadly, malignancy of the human gut (5-year survival &amp;lt;8%). NM is a global hotspot for GBC. Native Americans (NA) living in New Mexico (NM) have a 5- to 8-fold higher incidence of GBC compared to Caucasians in NM. The risk factors driving GBC incidence disparities in NM are unknown. We hypothesize that heavy metal exposures such as cadmium (Cd) and uranium (U) may play a key role in gallbladder carcinogenesis in NM due to the abundance of these metals in the local New Mexican environment. Methods and Procedures: We established a retrospective clinical cohort of 24 GBC patients in NM stratified by ethnicity. Archived FFPE samples were sequenced using DNA-seq approaches to identify key mutational patterns of GBC in NM. Key changes were validated via Immunohistochemistry and FISH approaches. Next, we used primary human GB cell lines to measure effects of Cd exposures on GB cell signaling activation. In particular, we focused on effects on Akt, MAPK, and COX-2 signaling. A combination of cell viability, quantitative reactive oxygen species (ROS), and Western blot-based molecular biology assays was used to understand Cd exposure effects on cell signaling dysregulation in a gallbladder epithelial cell model. Results: DNA-seq analysis results show the mutational spectrum of GBC samples in NM for the first time. We observed an elevated incidence of mutations in the Akt and MAPK gene pathways similar to results from earlier studies in GBC patients in China. TP53 was the commonest mutation seen in the NM GBC cohort (36%). Of importance, we observed that approximately 25% of NM GBC patients have a targetable HER2 amplification (n=6/24). In our GB epithelial cell studies, we observed a dose-dependent reduction of cell viability of gallbladder epithelial cells due to Cd exposures. The measured EC50 values of gallbladder primary cells range from 11-18 μM. Importantly, we observed activation of the Akt and MAPK signaling pathways via increased phosphorylation. The ROS activity in GB epithelial cells is elevated due to Cd exposures, which are responsible for activation of Akt and MAPK pathways. We also observed that Cd exposures led to elevation of the COX-2 expression, supporting its role in chronic GB inflammation that is characteristic of gallbladder carcinogenesis. Conclusions: While there is epidemiologic evidence for a role for heavy metal exposures in GBC, the basic research evidence supporting this hypothesis is lacking thus far. Our study shows, for the first time, a definitive role for cadmium exposures in GB cell signaling activation via the Akt and MAPK signaling pathways. Additionally, Cd exposure may also lead to COX-2 activation, which is a second key feature that explains the chronic inflammatory origins of GBC. Thus, our initial studies demonstrate a role for environmental heavy metal exposures (Cd) in gallbladder carcinogenesis for the first time. Future research will seek to strengthen the role of Cd and U in particular in GB carcinogenesis in NM. Citation Format: Rama R. Gullapalli, Trevar S. Caldwell, Megan N. Rivera, Priyanka Sharma. Gallbladder cancer disparities in New Mexico: Examining the role of environmental heavy metal exposures as a driver of gallbladder epithelial signaling dysfunction [abstract]. In: Proceedings of the AACR Special Conference on Environmental Carcinogenesis: Potential Pathway to Cancer Prevention; 2019 Jun 22-24; Charlotte, NC. Philadelphia (PA): AACR; Can Prev Res 2020;13(7 Suppl): Abstract nr A10.

Spectrum of computed tomography findings in gall bladder carcinoma patients: a retrospective observational study from tertiary care oncology setup

Background: Gallbladder carcinoma (CA) is the most common primary biliary carcinoma and fifth most common malignancy of the gastrointestinal tract. Patients are usually asymptomatic early in the course of the disease and as a result, therapeutic window is usually missed. Authors discuss spectrum of imaging findings in CA gall bladder with multidetector computed tomography (CT) as modality of choice.Methods: Retrospective cross-sectional study conducted at Shaukat Khanum memorial hospital and research centre. Data from January 2019 to January 2020 which included 70 patients was collected from HIS. Patients with imaging findings of CA gall bladder were examined who had undergone contrast enhanced CT. Institutional review board approved this study.Results: Out of 70 cases in this study, cholelithiasis was present in 60%. The commonest MDCT finding was mass replacing GB (78.6%), followed by eccentric/diffuse gall bladder wall thickening. Extension to porta hepatis was noted in 25 (35.7%) cases. Antroduodenal involvement was noted in 11 (15.8%), hepatic flexure involvement in 8 (11.4%). Vascular encasement/thrombosis of the portal vein noted in 6 (8.6%). Metastasis at presentation was seen in 16 patients i.e., 22.9%. Majority patients presented with stage III and IV disease.Conclusions: Single-center tertiary care oncology hospital study gives insight about imaging findings of carcinoma gallbladder. Authors conclude that multidetector CT is the diagnostic tool of choice in detection of gall bladder carcinoma, locoregional disease, distant metastasis and hence operability and non-operability of the disease.

Computer-aided assessment of the chemokine receptors CXCR3, CXCR4 and CXCR7 expression in gallbladder carcinoma.

Gallbladder carcinoma (GBC) is a vicious and invasive disease. The major challenge in the clinical treatment of GBC is the lack of a suitable prognosis method. Chemokine receptors such as CXCR3, CXCR4 and CXCR7 play vital roles in the process of tumour progression and metastasis. Their expression levels and distribution are proven to be indicative of the progression of GBC, but are hard to be decoded by conventional pathological methods, and therefore, not commonly used in the prognosis of GBC. In this study, we developed a computer‐aided image analysis method, which we used to quantitatively measure the expression levels of CXCR3, CXCR4 and CXCR7 in the nuclei and cytoplasm of glandular and interstitial cells from a cohort of 55 GBC patients. We found that CXCR3, CXCR4 and CXCR7 expressions are associated with the clinicopathological variables of GBC. Cytoplasmic CXCR3, nuclear CXCR7 and cytoplasmic CXCR7 were significant predictive factors of histology invasion, whereas cytoplasmic CXCR4 and nuclear CXCR4 were significantly correlated with T and N stage and were associated with the overall survival and disease‐free survival. These results suggest that the quantification and localisation of CXCR3, CXCR4 and CXCR7 expressions in different cell types should be considered using computer‐aided assessment to improve the accuracy of prognosis in GBC.

Combination of CA19-9 and the Neutrophil-to-Lymphocyte Ratio for the Differential Diagnosis of Gallbladder Carcinoma.

PurposeThis study aimed to evaluate the efficiency of several parameters, including the neutrophil-to-lymphocyte ratio (NLR) obtained from preoperative routine blood examination, either alone or as an adjunct to the carbohydrate antigen 19-9 (CA19-9), for the diagnosis of gallbladder carcinoma (GBC).Patients and MethodsData from 123 patients with gallbladder cholesterol polyps (GCP), 80 with gallbladder adenoma (GA), and 103 with GBC were reviewed retrospectively. Receiver operating characteristic analysis was used to assess the sensitivity, specificity, and clinical value of the NLR, CA19-9, and their combination.ResultsValues of measured indicators were significantly higher in GBC patients than in GCP or GA patients but there were no significant differences between GCP and GA patients. The combination of NLR and CA19-9 had the best diagnostic efficiency for differentiating GBC from benign lesions with 74.8% sensitivity and 89.7% specificity. However, the NLR showed no significant difference between mid-to-advanced stage and early-to-mid stage GBC. The combination of NLR and CA19-9 (53.7% sensitivity and 88.9% specificity) did not reveal any advantages over CA19-9 alone (63.0% sensitivity and 89.0% specificity) in distinguishing different stages of GBC.ConclusionNLR and CA19-9, and their combination―parameters that are easily obtained preoperatively―have potential as diagnostic markers for GBC.

RNA Sequencing Revealed Signals of Evolution From Gallbladder Stone to Gallbladder Carcinoma.

Gallbladder stone is a major risk factor for gallbladder carcinoma (GBC), while there is still a controversy whether period of follow-up since newly diagnoses of asymptomatic gallstones increases the risk of GBC. In this study, 10 GBC patients and 30 patients with gallstones were admitted to our hospital. Patients with gallstones were divided into 3 groups according to the follow-up time, involving 10 patients with follow-up period of 1–3 years (GS3 group), 10 patients with follow-up period of 5–10 years (GS5 group), and 10 patients with follow-up period of more than 10 years (GS10 group). Tumor and para-tumor tissues of GBC patients, and gallbladder tissues of gallstone patients were collected. RNA sequencing was performed on the 50 samples. Besides, 1,704 differentially expressed genes (DEGs) were identified in tumors compared with para-tumor tissues of 10 GBC patients, which were enriched into some well-known cancer-related pathways, such as PI3K-Akt, mitogen-activated protein kinase (MAPK), Ras, and Wnt signaling pathways, and the most significant pathway was neuroactive ligand-receptor interaction. Patients with gallstones with periods of follow-up equal to 1–3 and > 10 years showed to have higher cancer risk than those with 5–10 years. ALPP and GPR87 are potential biomarkers for predicting cancer risk in patients with gallstones. The in vitro results revealed that GPR-87 can promote the proliferation, migration, and invasion of GBC cells. Herein, we explored the relationship between GBC patients and patients with gallstones with different periods of follow-up in transcriptome level.