Prognostic gene expression signature for high-grade serous ovarian cancer.

BACKGROUND: Median overall survival (OS) for women with high-grade serous ovarian cancer (HGSOC) is ∼4 years, yet survival varies widely between patients. There are no well-established, gene expression signatures associated with prognosis. The aim of this study was to develop a robust prognostic signature for OS in patients with HGSOC. PATIENTS AND METHODS: Expression of 513 genes, selected from a meta-analysis of 1455 tumours and other candidates, was measured using NanoString technology from formalin-fixed paraffin-embedded tumour tissue collected from 3769 women with HGSOC from multiple studies. Elastic net regularization for survival analysis was applied to develop a prognostic model for 5-year OS, trained on 2702 tumours from 15 studies and evaluated on an independent set of 1067 tumours from six studies. RESULTS: Expression levels of 276 genes were associated with OS (false discovery rate < 0.05) in covariate-adjusted single-gene analyses. The top five genes were TAP1, ZFHX4, CXCL9, FBN1 and PTGER3 (P < 0.001). The best performing prognostic signature included 101 genes enriched in pathways with treatment implications. Each gain of one standard deviation in the gene expression score conferred a greater than twofold increase in risk of death [hazard ratio (HR) 2.35, 95% confidence interval (CI) 2.02-2.71; P < 0.001]. Median survival [HR (95% CI)] by gene expression score quintile was 9.5 (8.3 to -), 5.4 (4.6-7.0), 3.8 (3.3-4.6), 3.2 (2.9-3.7) and 2.3 (2.1-2.6) years. CONCLUSION: The OTTA-SPOT (Ovarian Tumor Tissue Analysis consortium - Stratified Prognosis of Ovarian Tumours) gene expression signature may improve risk stratification in clinical trials by identifying patients who are least likely to achieve 5-year survival. The identified novel genes associated with the outcome may also yield opportunities for the development of targeted therapeutic approaches.

Histopathological growth patterns of colorectal liver metastasis exhibit little heterogeneity and can be determined with a high diagnostic accuracy.

Colorectal liver metastases (CRLM) exhibit distinct histopathological growth patterns (HGPs) that are indicative of prognosis following surgical treatment. This study aims to assess the reliability and replicability of this histological biomarker. Within and between metastasis HGP concordance was analysed in patients who underwent surgery for CRLM. An independent cohort was used for external validation. Within metastasis concordance was assessed in CRLM with ≥ 2 tissue blocks. Similarly, concordance amongst multiple metastases was determined in patients with ≥ 2 resected CRLM. Diagnostic accuracy [expressed in area under the curve (AUC)] was compared by number of blocks and number of metastases scored. Interobserver agreement (Cohen's k) compared to the gold standard was determined for a pathologist and a PhD candidate without experience in HGP assessment after one and two training sessions. Both the within (95%, n = 825) and the between metastasis (90%, n = 363) HGP concordance was high. These results could be replicated in the external validation cohort with a within and between metastasis concordance of 97% and 94%, respectively. Diagnostic accuracy improved when scoring 2 versus 1 blocks(s) or CRLM (AUC = 95.9 vs. 97.7 [p = 0.039] and AUC = 96.5 vs. 93.3 [p = 0.026], respectively), but not when scoring 3 versus 2 blocks or CRLM (both p > 0.2). After two training sessions the interobserver agreement for both the pathologist and the PhD candidate were excellent (k = 0.953 and k = 0.951, respectively). The histopathological growth patterns of colorectal liver metastasis exhibit little heterogeneity and can be determined with a high diagnostic accuracy, making them a reliable and replicable histological biomarker.

[Merging different biobanks under one roof : Benefits and constraints on the way to a centralized biobank using the example of the BMBH]. / Alle unter einem Dach : Erfolge und Herausforderungen auf dem Weg zu einer zentralisierten Biobank am Beispiel der BMBH.

Founded in 1386, Heidelberg University is Germany's oldest and one of Europe's most reputable universities. As a scientific hub in Germany, Heidelberg is home to several internationally renowned medical research facilities that have an enormous demand for biomaterial samples and data-especially in the field of translational and cancer research.The main objective of the BMBF-funded project "BioMaterialBank Heidelberg" (BMBH) was the harmonization of local biobanking under the same administrative roof through the implementation of common and standardized project, data, and quality management procedures.In the very beginning, existing structures and processes of the participating biobanks in Heidelberg were identified and a common administrative structure with central representatives for IT and quality management (QM) was established to coordinate all BMBH activities.Over time, implementation of consented structures and processes took place, also revealing organizational challenges that had to be solved concerning, for example, differences in sample handling and the definition of consistent access regulations.We will discuss below these challenges as well as the opportunities of building a centralized biobank and show how issues can be resolved using the example of the BMBH.

[Preanalytics and biobanking : Influence of preanalytical factors on tissue sample quality]. / Präanalytik und Biobanking : Einfluss präanalytischer Faktoren auf die Gewebeprobenqualität.

Access to well-characterized human biosamples is one of the most important prerequisites for modern biomedical research. Biobanks play a decisive role here, as they provide corresponding biosamples for planned analyses. Many interfering factors influencing the quality of biosamples have to be taken into account. In addition to logistical, ethical, and data protection aspects, preanalytical variables in the context of sample acquisition, storage, and processing should be mentioned in particular. In this paper, therefore, the most important preanalytical influencing factors are presented systematically and an overview of current national and international activities for the standardized recording of these factors is provided with the goal of being able to better understand their influence on results and to minimize them in the near future.

Challenges for quality management in implementation, maintenance, and sustainability of research tissue biobanks.

Availability of high-quality human tissue samples and access to associated histopathological and clinical data is essential for basic and translational biomedical research, especially in areas of personalized medicine, drug, and biomarker development and mechanistically oriented biomedical research projects. Therefore, it is pivotal to establish and maintain quality-assured tissue biobanks that provide high-quality biomaterial to research thereby increasing the impact and reliability of scientific results. Quality concerns do not only address the biomaterial specimen itself but include all biobanking-related procedures. Tissue biobanks thus face essential challenges that encompass the implementation of adequate structural components, documentation of tissue sample collection and storage (procedures), as well as data and project management and IT. An integral and indispensable component of tissue biobanks is expert-driven evaluation (entry and exit controls) of tissue specimen to guarantee provision of high-quality assured biomaterials.

[Maintainance of a research tissue bank. (Infra)structural and quality aspects]. / Aufbau und Betrieb einer Gewebebank. (Infra)strukturelle und qualitative Herausforderungen.

The availability of high quality human tissue samples and access to associated histopathological and clinical data are essential for biomedical research. Therefore, it is necessary to establish quality assured tissue biobanks that provide high quality tissue samples for research purposes. This entails quality concerns referring not only to the biomaterial specimen itself but encompassing all procedures related to biobanking, including the implementation of structural components, e.g. ethical and legal guidelines, quality management documentation as well as data and project management and information technology (IT) administration. Moreover, an integral aspect of tissue biobanks is the quality assured evaluation of every tissue specimen that is stored in a tissue biobank and used for projects to guarantee high quality assured biomaterial.

[Structure of biobanks for urological research]. / Struktur von Biobanken für die urologische Forschung.

Biomedical research plays an important role in the development of novel diagnostic procedures, drugs and treatment strategies with regard to cancerous and chronic inflammatory diseases. Biobanks are essential tools in this process. The complex structures and benefits of biobanks are presented in this article.

Frequency of therapy-relevant staging shifts in colorectal cancer through the introduction of pN1c in the 7th TNM edition.

BACKGROUND: pN1c is a novel N-category introduced for colorectal cancer (CRC) in current TNM (Tumour, Node, Metastasis) classification. It represents cancers displaying tumour deposits (TDs) in the fat but no involvement of lymph nodes. pN1c is integrated into the UICC (International Union Against Cancer) staging system and shifts previous stage II cancers (6th edition) to stage III. We investigated the frequency of upstaging and TD prognostic significance. METHODS: 414 CRCs, consecutively collected during a population-based epidemiological study, TNM classified and UICC staged according to the 6th TNM edition were reinvestigated for TD presence. The association with survival was investigated after a median follow-up time of 5years in multivariate analyses among nodal negative and positive cases. RESULTS: TDs were found in 103 (24.9%) cancers and were strongly associated with T-, N- and M-stages (p<0.0001, each). Upstaging of previous stage II cancers by the presence of TDs (pN1c) was found in six of 140 cases (4.3% of stage II, 1.4% of all tumours). For stage III CRC, strongly reduced overall, CRC-specific and recurrence-free survival were observed with the presence of TDs (hazard ratios (HR) 2.29, 95% confidence interval 1.27-4.10, HR 2.51, 1.27-4.98, and HR 2.43, 1.32-4.48, respectively). CONCLUSIONS: Upstaging of CRCs through the introduction of pN1c occurs in less than 5% of previous stage II and less than 2% of all cancers. Given the biologic relevance of TDs, integration into the UICC staging relevant N-category is justified. The high prognostic impact of TDs, however, is not reflected in nodal positive cancers in both the TNM and UICC staging systems.

Expression of oestrogen receptor ß and prognosis of colorectal cancer.

BACKGROUND: Previous studies suggest that sex steroids influence colorectal cancer (CRC) carcinogenesis. The oestrogen receptor ß (ERß) is the predominantly expressed ER in the colon and loss of ERß in CRC has been associated with advanced cancer stages. METHODS: Information on vital status by the end of 2009 was obtained for 1262 CRC patients recruited between 2003 and 2007. The ERß expression was immunohistochemically measured and associations of ERß scores with overall survival (OS), disease-specific survival (DSS) and disease-free survival (DFS) were evaluated using Cox proportional hazard models adjusted for prognostic factors, such as tumour stage and second primary tumours. RESULTS: Of the 1101 tumour samples with successful measurement, 535 were ERß negative (48.6%), 381 (34.6%) showed moderate and 185 (16.8%) showed high ERß expression. Compared with high ERß expression, lack of ERß was associated with higher cancer stages as well as greater tumour extent. In multivariate analyses, ERß negativity was associated with an increased hazard ratio for death (HR=1.61, 95% CI 1.09-2.40, P=0.02), death attributed to CRC (HR=1.54, 95% CI 0.99-2.39, P=0.06) as well as a poorer DFS (DFS HR=1.64, 95% CI 1.23-3.36, P=0.04). The associations were stronger in stage I-III patients (OS HR=2.20, 95% CI 1.28-4.06, P=0.007, DSS HR=2.38, 95% CI 1.20-5.39, P=0.02, respectively). CONCLUSIONS: Lack of ERß expression is associated with advanced cancer stages and independently associated with poor survival.

Structural requirements of research tissue banks derived from standardized project surveillance.

Tissue banks constitute decisive and rate-limiting resource and technology platforms for basic and translational biomedical research, notably in the area of cancer. Thus, it is essential to plan and structure tissue banking and allocate resources according to research needs, but essential requirements are still incompletely defined. The tissue bank of the National Center of Tumor Diseases Heidelberg (NCT) was founded with the intention to provide tissues of optimal quality and to prioritize the realization of research projects. We analysed its structure and prospective project management registration as well as tracking records for all projects of the NCT tissue bank as of its start in 2005 in order to obtain information that may be relevant for tissue bank planning. All project proposals submitted to the NCT tissue bank (n = 681) were included in the study. For a detailed evaluation of provided services, only projects that were completed until July 2011 (n = 605) were analysed. For these 605 projects, NCT tissue bank provided 769 specific services. In all projects/services, we recorded project leader, type and amount of material provided, type of research (basic/translational), work load of project and project completion. Furthermore, all completed projects were tracked after 90 days according to a standard protocol to determine principal investigators' (PI) satisfaction and quality of the provided material. Until July 2011, 605 projects had been successfully completed as documented by material transfer agreement. Of the projects, 72.7 % addressed basic research, 22.3 % were translational research projects and 3 % concerned epidemiological research; 91 % (n = 546) concerned a single PI and the NTC tissue bank. For these projects, 769 specific services were provided. Of these services, 288 concerned providing formalin-fixed and paraffin-embedded (FFPE) tissue (extracts, full size sections), 126 providing fresh frozen materials (including fresh frozen sections), 137 providing tissue micro-array (TMA)-based sections and 199 providing immunohistochemical services. Project tracking demonstrated that all projects had started within 90 days after reception of the material by the PIs, and PI satisfaction with provided material exceeded 97 %. Standardized registration and tracking provides valuable structural information for planning and financing of tissue banks and allocation of resources. The high number of completed projects as well as high user satisfaction demonstrates that structuring of tissue banks should be preferably research-oriented and highly efficient. The comparable number of requests for FFPE and fresh frozen tissue as well as TMA-based services underpins the need for a broad approach in terms of methods and material types in order to fulfil research needs.

Epigenetic screen of human DNA repair genes identifies aberrant promoter methylation of NEIL1 in head and neck squamous cell carcinoma.

Aberrant promoter methylation of different DNA repair genes has a critical role in the development and progression of various cancer types, including head and neck squamous cell carcinomas (HNSCCs). A systematic analysis of known human repair genes for promoter methylation is however missing. We generated quantitative promoter methylation profiles in single CpG units of 160 human DNA repair genes in a set of DNAs isolated from fresh frozen HNSCC and normal tissues using MassARRAY technology. Ninety-eight percent of these genes contained CpG islands (CGIs) in their promoter region; thus, DNA methylation is a potential regulatory mechanism. Methylation data were obtained for 145 genes, from which 15 genes exhibited more than a 20% difference in methylation levels between tumor and normal tissues, manifested either as hypermethylation or as hypomethylation. Analyses of promoter methylation with mRNA expression identified the DNA glycosylase NEIL1 (nei endonuclease VIII-like 1) as the most prominent candidate gene. NEIL1 promoter hypermethylation was confirmed in additional fresh frozen HNSCC samples, normal mucosa, HNSCC cell lines and primary human skin keratinocytes. The investigation of laser-microdissected tissues further substantiated increased methylation levels in tumor versus matched non-tumor cells. Immunohistological analysis revealed significantly less NEIL1 protein expression in tumor tissues. 5-Aza-2'-deoxycytidine treatment and DNMT1 knockdown resulted in the re-expression of NEIL1 in HNSCC cell lines, which initially carried hypermethylated promoter regions. In conclusion, our results suggest that DNA methylation contributes to the downregulation of NEIL1 expression and might thus have a role in modulating the response to therapies of HNSCC.

[S100 protein positive sustentacular cells in pulmonary carcinoids and thoracic paragangliomas: differential diagnostic and prognostic evaluation]. / S100-Protein-positive Sustentakularzellen in pulmonalen Karzinoiden und thorakalen Paragangliomen : Differenzialdiagnostische und prognostische Bedeutung.

Paragangliomas have a classical histomorphology comprising a so-called "Zellballen" or nesting pattern with surrounding S100 protein positive sustentacular cells (SC) which form a meshwork with a wire-fence appearance. In adrenal and extra-adrenal paragangliomas the prevalence of SC is inversely associated with the patients' outcome. In order to get more insight into the prevalence as well as the prognostic and differential diagnostic value of this cell population in pulmonary carcinoids, we investigated a panel of 26 tumorlets, 147 typical and atypical pulmonary carcinoids and ten thoracic paragangliomas immunohistochemically. We were able to demonstrate that S100 protein positive cells are similarly distributed in both thoracic paragangliomas and pulmonary carcinoids. Hence, the presence and distribution of these cells does not appear to represent a reliable criterion in differential diagnosis. Moreover, all pulmonary carcinoid patients with a worse outcome had low numbers of or no S100 protein positive cells in their tissue specimens. Thus, the prevalence of these cells may potentially aid in prognostic assessment of pulmonary carcinoids, especially in biopsies.

Identification of immunohistochemical prognostic markers for survival after resection of pulmonary metastases from colorectal carcinoma.

BACKGROUND: Although aggressive resection of pulmonary metastases prolongs the survival of patients with metastatic colorectal cancer, there is a need for predictive pathologic parameters to understand the key molecular events of metastatic progression. The aim of this study was to verify immunohistochemical markers in addition to established clinical parameters after surgery. METHODS: From our subset of patients undergoing resection of pulmonary metastases from metastatic colorectal carcinoma, we analyzed 39 patients (23 men and 16 women) between 2003 and 2007. Only patients who met the criteria for a potentially curative operation were included. All patients were analyzed with regard to age and sex, primary tumor location, stage of the primary tumor, history of hepatic metastases, number of pulmonary metastases, pre-thoracotomy carcinoembryonic (CEA) serum antigen level, and the presence of thoracic lymph node metastasis. Furthermore, we immunohistochemically investigated the expression of vascular endothelial growth factor (VEGF)-D, FBJ murine osteosarcoma viral oncogene homolog B (FOS-B), and melanoma antigen (MAGE)-A in the surgical specimens of pulmonary metastatic lesions. RESULTS: The overall 3-year survival was 50.6 %. A significantly longer survival was observed with multivariate analysis in patients with a pre-thoracotomy serum carcinoembryonic antigen level of no more than 4.2 ng/mL ( P = 0.001), and Dukes stage A or B primary tumor ( P = 0.001). A significantly longer recurrence-free survival was observed with multivariate analysis in patients without thoracic lymph node involvement compared to patients with pulmonary and/or mediastinal lymph node metastases ( P = 0.006). The stage of the primary tumor remained significant ( P = 0.029), and FOS-B expression in tumor cells showed a trend towards favorable recurrence-free survival after pulmonary metastasectomy ( P = 0.059). No statistically significant difference was found in the overall survival rate or recurrence-free survival rate of patients with expression of VEGF-D or MAGE-A antigen in pulmonary metastatic tumor cells. CONCLUSIONS: Our results suggest that in addition to clinically prognostic factors, FOS-B expression has a debatable impact on patient survival. We conclude that the evaluation of molecular and clinical prognostic parameters at the time of pulmonary metastasectomy offers a greater understanding of the metastatic process and provides important information for patient selection.

[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.

[Endovascular repair of aspergilloma-induced arrosion bleeding of the subclavian artery]. / Endovaskuläre Therapie einer aspergilloseinduzierten septischen Arrosionsblutung der A. subclavia.

Invasive mycotic infections like aspergillosis are a major cause of death in patients with malignant haematologic diseases. Autopsy studies show that every fourth patient suffers from systemic mycotic infections. The most common cause of bleeding in aspergillosis is haemoptysis due to lung involvement, but also visceral organs like liver and kidneys are infested. We report a case of aspergillosis in which, apart from initial liver and kidney symptoms, septic arrosion of the subclavian artery also appeared. The case is described with special emphasis on the possibility of endovascular treatment as a bridging method for this fulminant complication.

An institutional study on thymomas and thymic carcinomas: experience in 77 patients.

BACKGROUND: Thymomas and thymic carcinomas are rare tumors of the anterior mediastinum. A WHO classification was introduced in 1999, which has been updated in 2004. Meanwhile, several retrospective studies have been carried out which have shown the prognostic significance of this classification together with Masaoka's staging system and the extent of surgery. PATIENTS AND METHODS: Between 1983 and 2000, 77 patients (37 male, 40 female) underwent resection of thymomas and thymic carcinomas in our institution. Complete resection was achieved in 57 patients. The median follow-up was 72.6 months. RESULTS: The overall 5-year survival rate was 71.4 %. The factors "histology" and "extent of resection" had the most important impact on survival. However, even among the patients with complete resection, 12 of them suffered a relapse. Among this patient group, the most important factors for disease-free survival were "tumor stage" and "histology". Patients with an incomplete resection had a 5-year survival rate of only 29 % in spite of adjuvant radiation and/or chemotherapy. Due to the high rate of relapse, the poor survival rate found in incompletely resected patients as well as the failure of classical chemotherapy regimens, especially in type B2 and type B3 thymomas and thymic carcinomas, the search for new chemotherapeutic schemes is mandatory. CONCLUSION: Our study shows that there are still encouraging therapeutic options for thymomas and thymic carinomas. Type B2, type B3 thymomas and thymic carcinomas have worse outcomes in spite of adjuvant chemo- and radiotherapies. Especially in patients with incomplete surgical resection the outcome remains poor.

Mediastinal angiomyolipomas in a male patient affected by tuberous sclerosis.

Classical angiomyolipomas are benign tumours composed of various tissues, including components of fat, abnormal blood vessels and smooth muscle cells. They are often found in association with tuberous sclerosis complex (TSC). The present study reports a male patient affected by TSC with intermittent, massive chylous pleural effusions, who developed recurrent mediastinal angiomyolipomas. The tumours were characterised via histological and immunohistochemical methods. Although angiomyolipomas frequently occur in the kidneys of TSC patients, this case is the first report of mediastinal angiomyolipomas associated with TSC. Besides lymphangioleiomyomatosis, this differential diagnosis has to be taken into account in the case of chylous pleural effusions and mediastinal masses in tuberous sclerosis complex patients.

Bilateral surgical resection in pulmonary epitheloid hemangioendothelioma.

Epitheloid hemangioendothelioma is a vascular tumour with an epitheloid appearance, originating from endothelial cells. Although it is a slow growing tumour, extensive pulmonary involvement, intrathoracic spread, and systemic spread have been documented. We present a case of epitheloid hemangioendothelioma of the lung in a patient with an initial diagnosis made by transthoracic biopsy. The prognosis is unpredictable, with life expectancy ranging from 1 to 20 years. There is no single effective treatment, though spontaneous regression and response to chemotherapy and interferon are reported. Our patient underwent pulmonary lobectomy of the right lower lobe and pulmonary wedge resection of the nodule located in the left lower lobe.