[From panel diagnostics to comprehensive genomic analysis : Infobesity or empowerment?] / Von der Paneldiagnostik zu umfassenden genomischen Analysen : Informationsüberfluss oder Zugewinn?

Precision oncology is obtaining a central role in the therapy of malignant diseases. The indication for targeted therapy is based on the identification of molecular targets for which next-generation sequencing (NGS) is commonly used nowadays. All approved predictive biomarkers and molecular targets, including gene fusions and copy number alterations, can be identified depending on panel design and method applied. Some clinical scenarios, however, may require more holistic genomic approaches, such as whole-genome/whole-exome and transcriptome analysis, which must be embedded in a clinical trial. Here, key aspects and applications of each method are summarized and discussed.

Inhibition of caspases primes colon cancer cells for 5-fluorouracil-induced TNF-α-dependent necroptosis driven by RIP1 kinase and NF-κB.

Resistance towards the drug 5-fluorouracil (5-FU) is a key challenge in the adjuvant chemotherapy of colorectal cancer (CRC), and novel targeted approaches are required to improve the therapeutic outcome. Necroptosis is a recently discovered form of programmed cell death, which depends on receptor interacting protein 1 (RIP1) and particularly occurs under caspase-deficient conditions. The targeted induction of necroptosis represents a promising strategy to overcome apoptosis resistance in cancer. The aim of this study was to systematically explore the usage of pan-caspase inhibitors to sensitize resistant CRC cells for 5-FU. We found that pan-caspase inhibitors facilitated 5-FU-induced necroptosis, which was mediated by autocrine secretion of tumor necrosis factor α (TNF-α). TNF-α production was driven by nuclear factor κB (NF-κB) and required RIP1 kinase. In vivo xenograft experiments showed that the novel pan-caspase inhibitor IDN-7314 in combination with 5-FU synergistically blocked tumor growth. Ex vivo experiments with fresh human CRC tissue specimens further indicated that a subgroup of patients could benefit from combinatory treatment. Thereby, elevated levels of secreted TNF-α and expression of components of the necroptotic pathway might help to predict the sensitivity to pro-necroptotic therapies. Together, our results shed new light on the molecular regulation of necroptosis by NF-κB and RIP1. Moreover, we identify necroptotic cell death as an important effector mechanism of 5-FU-mediated anti-tumoral activity. On the basis of this study, we propose pan-caspase inhibitors as a novel approach in the adjuvant chemotherapy of CRC.

[Molecular pathology of sarcomas. Update on the research group "Molecular Diagnosis of Sarcomas"]. / Molekularpathologie von Sarkomen. Erste Ergebnisse des Sarkomforschungsverbundes KoSar.

To establish precise diagnostic algorithms and standardised treatment of sarcomas in specialized centers, the interdisciplinary research group KoSar (sarcoma competence network) has been funded by German Cancer Aid. A sarcoma tissue repository and a diagnostic reference center have been set up, presently containing about 1000 accurately diagnosed sarcomas of different entities. Significant gene expression profiles for synovial sarcomas, leiomyosarcomas, myxoid liposarcomas and a small profile for myxofibrosarcomas as well as a new classification of angiosarcomas were defined. We systematically searched for activated signal transduction pathways in sarcoma cell lines and xenograft transplant models and candidate targets for molecular therapies were identified. Based on these results first clinical studies have been initiated by the German Interdisciplinary Sarcoma Study Group (GISG).

[Tissue bank of the National Centre for Tumour Disease. An innovative platform for translational tumour]. / Gewebebank des Nationalen Zentrums für Tumorerkrankungen. Innovative Plattform für die translationale Tumorforschung.

The tissue bank of the National Centre for Tumour Diseases (NCT) in Heidelberg, Germany, was founded in 2005 by the University Hospital of Heidelberg and the German Cancer Research Centre as a section of the NCT. It is a nonprofit organization with a completely evaluated legal and ethical framework and supports the Comprehensive Cancer Centre concept. Its main aim is the acquisition and characterization of fresh-frozen and paraffin-embedded human tissues according to the standards of good scientific practice and the promotion of interdisciplinary tumour research of the comprehensive cancer centre and its cooperating partners. It also offers expert project assistance: a project leader can submit a short proposal, and the tissue collecting/preparing process will be performed in cooperation with a specialised pathologist and, if applicable, an experienced clinical researcher. The tissue bank is also a central platform for further developing of innovative technologies for tissue handling, e.g. multi-tissue-array and virtual microscopy, with links to digital image analysis and bioinformatics. Thus, the NCT tissue bank represents a model for innovative biobanking and for institutions with active interdisciplinary cancer research.

Vitamin A-induced cholestatic hepatitis: a case report.

We report a case of intrahepatic cholestasis due to chronic vitamin A supplementation. A 70-year-old woman was admitted to the hospital for jaundice and reduced nutritional and general status with a 2-month history of increasing cholestasis. Some years previously she had suffered from breast and ovarian cancer with subsequent surgery and chemotherapy. Chemotherapy was terminated one month before elevated serum transaminase activities and cholestatic serum markers were noted. Following the chemotherapy, supportive care included weekly vitamin A injections (100,000 IU per injection). Liver biopsy showed an acute toxic liver injury with focal parenchymal necrosis, sinusoidal lesions, inflammatory infiltrate (round cells, macrophages), and activation and proliferation of stellate cells. The hepatic vitamin A concentration was found to be significantly elevated. There were no signs of intrahepatic metastasis or liver cirrhosis. Treatment with ursodeoxycholic acid rapidly improved the cholestasis and led to a total recovery after three weeks.

Oval cell lines OC/CDE 6 and OC/CDE 22 give rise to cholangio-cellular and undifferentiated carcinomas after transformation.

BACKGROUND: There is compelling evidence for a parenchymal origin of the predominant cell lineage leading from preneoplastic clear and acidophilic glycogen storage foci through mixed and basophilic cell populations to hepatocellular carcinomas in the rat. However, a controversial question remains to be answered: Do the basophilic cell foci invariably originate from parenchymal cells or do oval cells also have the potential to give rise to this type of focus and progress to hepatocellular neoplasms? Oval cells are nonparenchymal epithelial cells with scant cytoplasm and ovoid nuclei that first appear in the periportal areas of the liver lobules and thereafter invade the whole parenchyma when animals are exposed to high doses of a wide range of chemical carcinogens. EXPERIMENTAL DESIGN: Two oval cell lines, OC/CDE 6 and OC/CDE 22, which had been established from rats fed a choline-deficient/DL-ethionine-supplemented diet for 6 or 22 weeks, were transformed either by leaving the cells in confluence for a long time period (OC/CDE 6) or by treating the cells with the alkylating agent N-methyl-N'-nitro-N-nitrosoguanidine. The transformed cells were injected subcutaneously in newborn rats and the tumors developing in these animals were analyzed histopathologically, ultrastructurally, and immunohistochemically. RESULTS: The two transformed oval cell lines gave rise to carcinomas, in which cholangiocellular, adenoid and solid tumor formations were observed. Subpopulations of these tumors expressed cytokeratins 7, 8, 18, and 19, but were albumin- and alpha-fetoprotein-negative. Areas within the carcinomas derived from transformed OC/CDE 22 cells representing undifferentiated liver tumor formations were also identified. Cells within these areas had lower nucleus/cytoplasm ratios than cells in the solid growing tumor formations, stained positive for cytokeratins 8 and 18 and were cytokeratin 7- and 19-, albumin- and alpha-fetoprotein-negative. Ultrastructurally, these cells did not resemble those of differentiated hepatocellular carcinomas. CONCLUSIONS: It has been shown that oval cells are precursor cells of carcinomas containing cholangiocellular, adenoid and solid formations which may be largely undifferentiated. However, the transformed OC/CDE 6 or OC/CDE 22 cells do not serve as precursor cells of differentiated hepatocellular carcinomas.