Thesis (M.S.)--University of Alabama at Birmingham, 2010. / Title from PDF title page (viewed on June 25, 2010). Includes bibliographical references (p. 31-37).

Bei ICCC (intrahepatischen cholangiozellulären Karzinomen) handelt es sich um eine seltene Erkrankung der Gallenwege. Die Erkrankung schreitet langsam voran und verläuft lange Zeit asymptomatisch oder verursacht nur unspezifische Symptome, sodass sich die meisten Patienten bei Diagnosestellung bereits in einem fortgeschrittenen, nicht-resezierbaren Tumorstadium befinden. Im Rahmen dieser retrospektiven Arbeit wurden Einflussfaktoren auf das Überleben und die Entwicklung eines Tumorrezidivs von Patienten untersucht, die aufgrund eines ICCC kurativ leberreseziert wurden. Hierbei wurden 42 Patienten eingeschlossen, die zwischen 2003 und 2015 am Universitätsklinikum Würzburg operiert wurden. Die Datenauswertung lieferte die Erkenntnis, dass sowohl das Vorhandensein multifokal lokalisierter ICCC als auch ein erhöhtes Lebensalter (60 Jahre und älter) zum Zeitpunkt der Operation mit einem geringeren perioperativen und langfristigen Überleben der Patienten assoziiert sind. Außerdem sind diese beiden Faktoren mit der Entstehung eines Tumorrezidivs, was die Überlebenszeit verkürzt, assoziiert. Hingegen konnte bei Auftreten von postoperativen Komplikationen und dem Vorhandensein einer höhergradigen ASA-Klassifikation (ASA III und ASA IV) eine Assoziation mit einem geringeren perioperativen nicht aber mit einem reduzierten langfristigen Überleben gezeigt werden. / ICCC (intrahepatic cholangiocarcinoma) is a rare disease of the biliary tract. The disease progresses slowly and is asymptomatic for a long time or causes only unspecific symptoms, so that most patients are already in an advanced, unresectable tumor stage. In the context of this retrospective study, factors influencing the survival and development of tumor recurrence in patients who were curatively liver resected due to ICCC were investigated. This included 42 patients who were operated between 2003 and 2015 at the Würzburg University Hospital. The data analysis revealed that both the presence of multifocal localized ICCC and an increased age (60 years and older) at the time of surgery are associated with lower perioperative and long-term survival of the patients. In addition, these two factors are associated with the development of tumor recurrence, which shortens survival time. On the other hand, postoperative complications and the existence of a higher-grade ASA classification (ASA III and ASA IV) are associated with a lower perioperative but not reduced long-term survival.

No description available.

Intrahepatic cholangiocarcinoma (ICC) is an aggressive malignancy with a dismal prognosis. Few patients present with disease amenable to resection and chemotherapy is not curative. The incidence of ICC is rising worldwide and new therapeutic approaches are urgently required. Notch signalling is critical for the embryological development and regeneration of the biliary tree in the mammalian liver. Dysregulation of Notch is known to drive tumorigenesis in a range of solid and haematological malignancies and the aim of this work was to define its contribution to the pathogenesis of ICC. Transgenic overexpression of Notch1 has been described to result in the formation of biliary lineage tumours in the liver. I have used resected human tissue, a chemically-induced model of ICC in rat and a novel transgenic murine model in which the tumour suppressor p53 is conditionally deleted from biliary epithelia, to demonstrate that endogenous Notch signalling is acting via the Notch3 receptor to drive tumorigenesis. I use multiple independent methods of Notch3 blockade to establish that Notch3 promotes epithelial cell survival and self-renewal in ICC and demonstrate that Notch3 inhibition significantly attenuates tumour growth in vivo. My data suggest that Notch3 promotes activity through the PI3K/AKT cell survival cascade via a mechanism independent of the effector of canonical Notch, RBPJκ. Given the significant toxicity associated with gamma-secretase inhibitors these findings offer a novel and specific target for further investigation and future therapeutic development in ICC.

No description available.

Le but de ce travail était de rechercher le rôle du microenvironnement dans la progression tumorale du cholangiocarcinome intrahépatique (CCIH) par une approche translationnelle, associant recherche fondamentale et clinique. Une étude transcriptomique du stroma tumoral a permis de mettre en évidence une signature spécifique de celui-ci, dont l’analyse non supervisée montrait un enrichissement dans les gènes de la matrice extracellulaire, du cycle cellulaire, de la voie TGFβ et des marqueurs de cellules souches. Ces résultats ont été validés au niveau protéique par immunohistochimie sur tissue microarrays à partir d’une cohorte indépendante. La corrélation de ces résultats avec les données cliniques a permis de démontrer que le niveau d’expression de l’Osteopontin dans le stroma était un facteur de risque indépendant de récidive et de survie. Par ailleurs, nous avons démontré que le taux sérique d’Osteopontin préopératoire des patients porteurs d’un CCIH était significativement supérieur à celui de sujets sains. Avec un seuil déterminé à 57,8 ng/ml, la sensibilité et spécificité de ce biomarqueur diagnostique était respectivement de 80 et 100%. De plus, nous avons apporté des arguments supplémentaires concernant le rôle des cellules souches cancéreuses dans la progression du CCIH, en mettant en évidence une corrélation entre le niveau d’expression de marqueurs souches tels qu’EpCAM et CD44 dans le stroma tumoral ainsi que dans le tissu fibreux du foie « sain » péri-lésionnel et le risque de récidive. Les résultats de notre étude ont confirmé le rôle central du microenvironnement dans la progression du CCIH, permis de mettre en évidence 2 nouveaux biomarqueurs pronostiques, et ouvert de nouvelles voies de recherche thérapeutiques. / The aim of this study was to specifically determine through a translational approach combining basic and clinical research, the role of the microenvironment in the tumor progression of intrahepatic cholangiocarcinoma (ICC). By gene expression profiling, we identified a signature that significantly discriminate the tumor stroma from non-tumor fibrous tissue, and the functional analysis of differentially expressed genes showed an enrichment in genes of the extracellular matrix , the cell cycle, the TGFb pathway and stem cell markers. Tissue microarray analysis using an independent cohort of ICC patients validated at a protein level the increased expression of selected candidate genes. Statistical analysis between basic and clinical data demonstrated that the stromal expression of Osteopontin was an independent prognostic marker for overall and disease-free survival. We also demonstrated that the preoperative serum level of Osteopontin was significantly higher in ICC patients than in healthy subjects. Our results identified the best diagnostic threshold to 57,8 ng/ml, associated with a sensitivity and specificity reaching to 80 and 100%, respectively. Moreover, we showed that level expression of stem cell markers such as EpCAM and CD44 in tumor stroma as well as in the fibrous non tumor liver tissue was correlated with recurrence, suggesting the pivotal role of cancer stem cells in ICC prognosis. In conclusion, our study confirmed the major involvement of the microenvironment in the progression of CCIH, allowed to identify two new prognostic tumor biomarkers, and highlighted new pathways for targeted therapeutics.

Background: Peroxisome proliferator-activated receptor δ (PPARδ) is a versatile regulator of distinct biological processes and overexpression of PPARδ in cancer may be partially related to its suppression of its own co-regulators. Aims: To determine whether recruited suppressor proteins bind to and regulate PPARδ expression, activity and PPARδ-dependent cholangiocarcinoma proliferation. Methods: Yeast two-hybrid assays were done using murine PPARδ as bait. PPARδ mRNA expression was determined by qPCR. Protein expression was measured by western blot. Immunohistochemistry and fluorescence microscopy were used to determine PPARδ expression and co-localization with NDP Kinase alpha (NM23-H2). Cell proliferation assays were performed to determine cell numbers. Results: Yeast two-hybrid screening identified NM23-H2 as a PPARδ binding protein and their interaction was confirmed. Overexpressed PPARδ or treatment with the agonist GW501516 resulted in increased cell proliferation. NM23-H2 siRNA activated PPARδ luciferase promoter activity, upregulated PPARδ RNA and protein expression and increased GW501516-stimulated CCA growth. Overexpression of NM23-H2 inhibited PPARδ luciferase promoter activity, downregulated PPARδ expression and AKT phosphorylation and reduced GW501516-stimulated CCA growth. Conclusions: We report the novel association of NM23-H2 with PPARδ and the negative regulation of PPARδ expression by NM23-H2 binding to the C-terminal region of PPARδ. These findings provide evidence that the metastasis suppressor NM23-H2 is involved in the regulation of PPARδ-mediated proliferation.

() is a commensal organism of the gastrointestinal tract. It rarely causes hepatobiliary infections; however, in the presence of underlying inflammation, immunosuppression, or malignancy, it can cause hepatobiliary infection via an ascending route. Herein, we report a case of pyogenic liver abscess secondary to associated with cholangiocarcinoma, which was treated with ceftriaxone and metronidazole.

MiR-17-92 is an oncogenic miRNA cluster implicated in the development of several human cancers; however, it remains unknown whether miR-17-92 cluster is able to regulate hepatobiliary carcinogenesis. This study was designed to investigate the biological functions and molecular mechanisms of miR-17-92 cluster in primary liver cancer.<br>In-situ hybridization and qRT-PCR analysis showed that miR-17-92 cluster is highly expressed in human cholangiocarcinoma cells compared to the non-neoplastic biliary epithelial cells. Forced overexpression of the miR-17-92 cluster or its members, miR-92a and miR-19a, in cultured human cholangiocarcinoma cells enhanced tumor cell proliferation, colony formation and invasiveness, in vitro. Overexpression of miR-17-92 cluster or miR-92a also enhanced cholangiocarcinoma growth in vivo in SCID hairless outbred mice. The tumor suppressor PTEN was identified as a bona fide target of both miR-92a and miR-19a in cholangiocarcinoma cells. Accordingly, overexpression of PTEN open reading frame protein (devoid of 3’UTR) prevented miR-92a- or miR-19a-induced cholangiocarcinoma cell growth. Microarray analysis revealed additional targets of miR-17-92 cluster in human cholangiocarcinoma cells, including APAF-1 and PRDM2. Moreover, we observed that the expression of miR-17-92 cluster is regulated by IL-6/Stat3, a key oncogenic signaling pathway pivotal in cholangiocarcinogenesis. Taken together, our findings in this study disclose a novel IL-6/Stat3 miR-17-92 cluster PTEN signaling axis that is crucial for cholangiocarcinogenesis and tumor progression.<br>We also found the miR-17-92 is highly expressed in tumor tissue compared to non-tumor adjacent tissue in hepatocellular carcinoma patient tissue. Forced overexpression of the miR-17-92 cluster in cultured human hepatocellular carcinoma cells enhanced tumor growth in vitro; on contrast, inhibition of miR-17-92 cluster inhibited cell growth. MiR-17-92 cluster promote diethylnitrosamine-induced hepatocarcinogenesis in liver-specific miR-17-92 cluster transgenic mice. Binding sequence and mice whole genome microarray analysis revealed about 300 possible targets. RNA-sequencing data analysis showed both individual miRNAs and the host gene of miR-17-92 cluster was highly expressed in hepatocellular carcinoma patients and had negative correlation with several genes (CREBL2, PRRG1, and NTN4), among which, CREBL2 may play an important role in the hepatocarcinogenesis. / acase@tulane.edu

MicroRNAs (miRNAs) are a group of small, noncoding RNAs that modulate the translation of genes into proteins by binding to specific target sites in messenger RNAs. This study investigated the biological function and molecular mechanism of microRNA-21 (miR-21) in human cholangiocarcinoma. In situ hybridization analysis of human cholangiocarcinoma tissues showed increased miR-21 in cholangiocarcinoma cells compared to the noncancerous biliary epithelial cells. Forced overexpression of miR-21 by lentivirus transduction enhanced human cholangiocarcinoma cell growth and clonogenic efficiency in vitro, whereas inhibition of miR-21 decreased these parameters. MiR-21 overexpression also promoted cholangiocarcinoma growth in a tumor xenograft model. The NAD+-linked 15-hydroxyprostaglandin dehydrogenase (15-PGDH), a key enzyme that converts the pro-tumorigenic prostaglandin E2 (PGE2) to biologically inactive metabolite, was identified as a direct target of miR-21 in cholangiocarcinoma cells. In parallel, cyclooxygenase-2 (COX-2) overexpression and PGE2 treatment increased miR-21 expression and enhanced miR-21 promoter reporter activity in human cholangiocarcinoma cells. Our results disclose a novel cross-talk between COX-2/PGE2 and miR-21 signaling pathways that converges at 15-PGDH which is crucial in cholangiocarcinogenesis and tumor progression. The enhancer of zeste homolog 2 (EZH2) is the catalytic subunit in the PRC2 complex catalyzing the trimethylation of histone3 lysine27 (H3K27) and mediates gene silencing of the target genes. The biological function of EZH2 in cholangiocarcinoma was investigated in this study. Immunohistochemistry staining of EZH2 on human cholangiocarcinoma tissues showed increased EZH2 expression in cholangiocarcinoma cells. Pharmacologically inhibition of EZH2 by EZH2 inhibitors decreased cholangiocarcinoma growth and induced G1 arrest. The CD133, one of the putative cancer stem cell markers, was found to express in the cholangiocarcinoma cell lines we used. Inhibition of EZH2 decreased CD133+ population and the sphere forming ability of cancer cells. Our results indicate that EZH2 may represent a promising target for targeting the tumor-initiating cell population and future cholangiocarcinoma therapy. / acase@tulane.edu