The beta-catenin-target Fascin-1, altering hepatocyte differentiation, is a new marker of immature cells in hepatoblastomas

OBJECTIVE: Beta-catenin is a well-known effector of the Wnt pathway and a key player in cadherin-mediated cell adhesion. Oncogenic mutations of beta-catenin are highly frequent in pediatric liver primary tumors. Those mutations are mostly heterozygous allowing the co-expression of wild-type (WT) and mutated beta-catenins in tumor cells. We investigated the interplay between WT and mutated beta-catenins in liver tumor cells, and searched for new actors of the beta-catenin pathway. DESIGN: Using an RNAi strategy in beta-catenin-mutated hepatoblastoma (HB) cells, we dissociated the structural and transcriptional activities of beta-catenin, carried mainly by, respectively, WT and mutated proteins. Their impact was characterized using transcriptomic and functional analyses. We studied mice that develop liver tumors upon activation of beta-catenin in hepatocytes (APCKO and beta-catenin∆exon3 mice). We used transcriptomic data from mouse and human HB specimens and analyzed samples by immunohistochemistry. RESULTS: We highlighted an antagonist role of WT and mutated beta-catenins on hepatocyte differentiation as attested by alteration of hepatocyte markers expression and bile canaliculi formation. We characterized Fascin-1 as a target of beta-catenin involved in hepatocyte differentiation. Using mouse models, we found that Fascin-1 is highly expressed in undifferentiated tumors. Finally, we found that Fascin-1 is a specific marker of the embryonal component in human HBs. CONCLUSION: In mice and human, Fascin-1 expression is linked to loss of differentiation and polarity of hepatocytes. Thus, we highlighted Fascin-1 as a new player in the modulation of hepatocyte differentiation associated to beta-catenin pathway alteration in the liver.

The Significant Impact of Immunohistochemistry in the Classification of Lung Carcinoma on Small Biopsies

Background : There are limitations of histomorphology in the appropriate categorization of lung carcinoma where immunohistochemistry can confirm the morphological diagnosis and may add value in the poorly differentiated and undifferentiated tumors. The aim of the study was to assess the role of immunohistochemistry in classifying lung carcinoma on small biopsy samples. Methods and Material: A retrospective hospital based, observational study was conducted on cases of lung carcinoma diagnosed by core needle or bronchoscopic biopsies over a 3- year period. After evaluation of clinical findings and H&E sections, all biopsies were evaluated by immunohistochemical staining. The immunohistochemistry panel included cytokeratin cocktail, CK7, CK 20, TTF1, Napsin A, CK5/6, p40, synaptophysin, chromogranin, CD 56. Result: Out of 78 cases, the mean age was 58 +/- 11 years. Most prevalent malignancy type was adenocarcinoma (30, 38.1%). Adenocarcinoma cases were positive for CK7 (25/26, 96%), Napsin A (24/26, 92%), TTF1 (15/30, 50%) and negative for CK 20. Squamous cell carcinoma cases showed positivity for p40(18/22, 82%) and CK 5/6 (17/22.71%).Small cell carcinoma showed positivity for neuroendocrine markers synaptophysin (5/7,71%) and chromogranin(4/7, 57%) and CD 56 (6/7cases (85%).88% of small cell carcinomas,75% of adenocarcinomas and 72 % of squamous cell carcinomas were accurately diagnosed by morphology. Morphologic prediction was poor in the NSCC NOS group (0%) and poorly differentiated carcinomas (64%), which were finally, diagnosed by immunohistochemistry.In the morphologically diagnosed cases, immunohistochemistry confirmed the diagnosis. Conclusion: Thus, morphology added with immunohisto chemistry provides a crisp diagnosis thereby improving therapeutic efficacy

Relationship of breast volume, obesity and central obesity with different prognostic factors of breast cancer

The objective of this study was to investigate whether the BC tumor biology in women with larger breast volume, in obese women and especially in women with central adiposity at the moment of diagnosis of BC is more aggressive than in those women without these characteristics. 347 pre- and postmenopausal women with a recent diagnosis of BC were analyzed. In all patients, anthropometric measurements at the time of diagnosis was collected. In 103 of them, the breast volume was measured by the Archimedes method. The Breast volume, BMI, WHR and the menopausal status were related to different well-known pathological prognostic factors for BC. At the time of diagnosis, 35.4% were obese (BMI > 30 kg/m2), 60.2% had a WHR ≥ 0.85, 68.8% were postmenopausal and 44.7% had a breast volume considered "large" (> 600 cc). Between patients with a large breast volume, only a higher prevalence of ER (+) tumors was found (95.3% vs. 77.2%; p = 0.04) compared to those with small breast volumes. The obese BC patients showed significantly higher rates of large tumors (45.5% vs. 40.6%; p = 0.04), axillary invasion (53.6% vs. 38.8%; p = 0.04), undifferentiated tumors (38.2% vs. 23.2%) and unfavorable NPI (p = 0.04) than non-obese women. Those with WHR ≥ 0.85 presented higher postsurgical tumor stages (61.7% vs. 57.8%; p = 0.03), higher axillary invasion (39.9% vs. 36.0%; p = 0.004), more undifferentiated tumors (30.0% vs. 22.3%; p = 0.009), higher lymphovascular infiltration (6.5% vs. 1.6%; p = 0.02), and a higher NPI (3.6 ± 1.8 vs. 3.2 ± 1.8; p = 0.04). No statistically significant differences were found according to menopausal status. We conclude that obesity, but especially central obesity can be associated with a more aggressive tumour phenotype. No relation between breast volume and tumoral prognostic factors was found, except for a higher proportion of ER (+) tumor in women with higher breast volume.

Ganglioneuroma as the phenomenon of neuroblastoma maturation

Ganglioneuroma (GN) represents a mature, well-differentiated tumor arising from the sympathetic nervous system. Mostly developing de novo, GN can appear during the treatment course of poorly differentiated or undifferentiated tumors of the sympathetic nervous system, such as neuroblastoma, or as a result of their spontaneous maturation. In this article we report three clinical cases of spontaneous and induced maturation of neuroblastoma (primary tumor and metastatic lesion) to GN. Histological verification of long-lasting stable or progressing residual tumor mases in patients with neuroblastoma stratified to the observation group plays a pivotal role as it may significantly affect the treatment course. The patients' parents gave their consent to the use of their child's data, including photographs, for research purposes and in publications.

Probiotic-Treated Super-Charged NK Cells Efficiently Clear Poorly Differentiated Pancreatic Tumors in Hu-BLT Mice

Background and Aims: We have previously demonstrated that the stage of differentiation of tumors has profound effect on the function of NK cells, and that stem-like/poorly differentiated tumors were preferentially targeted by the NK cells. Therefore, in this study we determined the role of super-charged NK cells in immune mobilization, lysis, and differentiation of stem-like/undifferentiated tumors implanted in the pancreas of humanized-BLT (hu-BLT) mice fed with or without AJ2 probiotics. The phenotype, growth rate and metastatic potential of pancreatic tumors differentiated by the NK cells (NK-differentiated) or patient derived differentiated or stem-like/undifferentiated pancreatic tumors were investigated. Methods: Pancreatic tumor implantation was performed in NSG and hu-BLT mice. Stage of differentiation of tumors was determined using our published criteria for well-differentiated tumors exhibiting higher surface expression of MHC- class I, CD54, and PD-L1 (B7H1) and lower expression of CD44 receptors. The inverse was seen for poorly-differentiated tumors. Results: Stem-like/undifferentiated pancreatic tumors grew rapidly and formed large tumors and exhibited lower expression of above-mentioned differentiation antigens in the pancreas of NSG and hu-BLT mice. Unlike stem-like/undifferentiated tumors, NK-differentiated MP2 (MiaPaCa-2) tumors or patient-derived differentiated tumors were not able to grow or grew smaller tumors, and were unable to metastasize in NSG or hu-BLT mice, and they were susceptible to chemotherapeutic drugs. Stem-like/undifferentiated pancreatic tumors implanted in the pancreas of hu-BLT mice and injected with super-charged NK cells formed much smaller tumors, proliferated less, and exhibited differentiated phenotype. When differentiation of stem-like tumors by the NK cells was prevented by the addition of antibodies to IFN-γ and TNF-α, tumors grew rapidly and metastasized, and they remained resistant to chemotherapeutic drugs. Greater numbers of immune cells infiltrated the tumors of NK-injected and AJ2-probiotic bacteria-fed mice. Moreover, increased IFN-γ secretion in the presence of decreased IL-6 was seen in tumors resected and cultured from NK-injected and AJ2 fed mice. Tumor-induced decreases in NK cytotoxicity and IFN-γ secretion were restored/increased within PBMCs, spleen, and bone marrow when mice received NK cells and were fed with AJ2. Conclusion: NK cells prevent growth of pancreatic tumors through lysis and differentiation, thereby curtailing the growth and metastatic potential of stem-like/undifferentiated-tumors.

LncRNAOIP5-AS1is overexpressed in undifferentiated oral tumors and integrated analysis identifies as a downstream effector of stemness-associated transcription factors

Long non-coding RNAs (lncRNAs) play an important role in the regulation of key cellular processes in early development and in cancer. LncRNAOip5-as1facilitates stem cell self-renewal in mouse by sponging mmu-miR-7 and modulating NANOG level, yet its role in cancer is less understood. We analyzedOIP5-AS1expression in oral tumors and in TCGA datasets. We observed overexpression ofOIP5-AS1in oral tumors (P<0.001) and in tumors of epithelial origin from TCGA.OIP5-AS1expression was strongly associated with undifferentiated tumors (P=0.0038).In silicoanalysis showed miR-7 binding site is conserved in mouse and humanOIP5-AS1. However, humanNANOG3’-UTR lost the binding site for hsa-miR-7a-3. Therefore, we screened for other miRNAs that can be sponged byOIP5-AS1and identified six potential miRNAs and their downstream target genes. Expression analysis showed downregulation of miRNAs and upregulation of downstream target genes, particularly in undifferentiated tumors with high-level ofOIP5-AS1suggesting thatOIP5-AS1could post-transcriptionally modulate the downstream target genes. Further, systematic epigenomic analysis ofOIP5-AS1promoter revealed binding motifs for MYC, NANOG and KLF4 suggesting thatOIP5-AS1could be transactivated by stemness-associated transcription factors in cancer. Overexpression of OIP5-AS1 in undifferentiated oral tumors may confer poor prognosis through maintenance of cancer stemness.

A population-based study of biliary tract cancers (BTCs) in Alberta, Canada: How do our patients fare?

246 Background: BTCs are poorly studied due to their rarity and heterogeneity. We explored demographics and outcomes of BTC pts over a 15 year period. Methods: All patients (pts) with biopsy-proven BTC (intrahepatic [IC] and extrahepatic [EC] cholangiocarcinomas, gallbladder cancers [GB], and ampulla of vater cancers [AV]) in Alberta were reviewed from January 1, 2000, to December 31, 2015. Demographic, pathologic, and survival data were extracted from electronic charts. Descriptive statistics were utilized. Overall survival (OS) was defined as the time from pathologic diagnosis to death date. Results: A total of 1,718 pts with BTCs were identified. Median age was 68 with 52% being male. The Table demonstrates OS breakdown based on tumour location and stage. Regardless of location of primary tumour, grade impacted survival (median OS in with well differentiated tumours vs undifferentiated tumors 26.6 vs 3.9 months). Pts who received standard of care palliative cisplatin/gemcitabine (Cis/Gem) chemotherapy (n = 123) had a median OS of 15.4 months. Conclusions: Patients with AV and IC have the best and worst prognosis, respectively. Shorter survival is observed with higher stage, grade, and unresectable disease. Pts who received palliative Cis/Gem had better OS than reported in the pivotal phase III trial. Further analysis of prognostic factors will be presented. [Table: see text]