TY - JOUR AB - BACKGROUND: We investigated the impact of SMAD2 linker phosphorylation (pSMAD2L) on overall and disease-free survival, signal transduction, as well as cancer-related processes in non-small cell lung cancer (NSCLC). METHODS: We generated A549 cells constitutively lacking pSMAD2L (A549Lsub) to gain mechanistic insights and stimulated NSCLC cell lines with inhibitors against cell cycle-associated kinases or TGFβ1. In addition, we analysed SMAD2 and pSMAD2L in alveolar epithelial cells type 2 from tumour-free lung tissue as well as in benign and malignant T cells by Western blotting. Furthermore, pSMAD2L-positive tumours and immune cells were analysed in an NSCLC patient cohort (n = 316) using multiplex immunofluorescence. RESULTS: In NSCLC cell lines and benign T cells, pSMAD2L was expressed in a mitosis-dependent manner. Loss of pSMAD2L (A549Lsub) had an anti-proliferative effect, slowed migration, and increased alternatively spliced short SMAD2 (SMAD2ΔE3). The gene signature in A549Lsub was associated with developmental and morphogenetic processes and redirected canonical TGFβ1-dependent signalling. By contrast, SMAD2ΔE3 was absent in benign T cells but present in malignant T lymphoblasts. NSCLC patients with low pSMAD2L+ tumour cell density had a poorer prognosis, whereas low pSMAD2L+ immune cell density favoured overall and disease-free survival. CONCLUSIONS: pSMAD2L antagonises anti-proliferative canonical TGFβ-signalling in NSCLC and redirects TGFβ1-dependent gene expression, whereas loss of pSMAD2L enhances SMAD2ΔE3 and affects pluripotency-associated proteins in vitro. In NSCLC patients, pSMAD2L cell density influences disease-free and overall survival in a spatially distinct manner. AU - Nitschkowski, D.* AU - Vierbuchen, T.* AU - Heine, H.* AU - Behrends, J.* AU - Reiling, N.* AU - Reck, M.* AU - Rabe, K.F.* AU - Kugler, C.* AU - Ammerpohl, O.* AU - Drömann, D.* AU - Muley, T.* AU - Kriegsmann, M.* AU - Stathopoulos, G.T. AU - Arendt, K.A.M. AU - Goldmann, T.* AU - Marwitz, S.* C1 - 74281 C2 - 57416 CY - Campus, 4 Crinan St, London, N1 9xw, England TI - SMAD2 linker phosphorylation impacts overall survival, proliferation, TGFβ1-dependent gene expression and pluripotency-related proteins in NSCLC. JO - Br. J. Cancer PB - Springernature PY - 2025 SN - 0007-0920 ER - TY - JOUR AB - BACKGROUND: Trastuzumab is the only first-line treatment targeted against the human epidermal growth factor receptor 2 (HER2) approved for patients with HER2-positive advanced gastric cancer. The impact of metabolic heterogeneity on trastuzumab treatment efficacy remains unclear. METHODS: Spatial metabolomics via high mass resolution imaging mass spectrometry was performed in pretherapeutic biopsies of patients with HER2-positive advanced gastric cancer in a prospective multicentre observational study. The mass spectra, representing the metabolic heterogeneity within tumour areas, were grouped by K-means clustering algorithm. Simpson's diversity index was applied to compare the metabolic heterogeneity level of individual patients. RESULTS: Clustering analysis revealed metabolic heterogeneity in HER2-positive gastric cancer patients and uncovered nine tumour subpopulations. High metabolic heterogeneity was shown as a factor indicating sensitivity to trastuzumab (p = 0.008) and favourable prognosis at trend level. Two of the nine tumour subpopulations associated with favourable prognosis and trastuzumab sensitivity, and one subpopulation associated with poor prognosis and trastuzumab resistance. CONCLUSIONS: This work revealed that tumour metabolic heterogeneity associated with prognosis and trastuzumab response based on tissue metabolomics of HER2-positive gastric cancer. Tumour metabolic subpopulations may provide an association with trastuzumab therapy efficacy. CLINICAL TRIAL REGISTRATION: The patient cohort was conducted from a multicentre observational study (VARIANZ;NCT02305043). AU - Wang, J. AU - Sun, N. AU - Kunzke, T. AU - Shen, J. AU - Feuchtinger, A. AU - Wang, Q. AU - Meixner, R. AU - Le Gleut, R. AU - Haffner, I.* AU - Luber, B.* AU - Lordick, F.* AU - Walch, A.K. C1 - 69867 C2 - 55293 CY - Campus, 4 Crinan St, London, N1 9xw, England SP - 1036-1045 TI - Metabolic heterogeneity affects trastuzumab response and survival in HER2-positive advanced gastric cancer. JO - Br. J. Cancer VL - 130 IS - 6 PB - Springernature PY - 2024 SN - 0007-0920 ER - TY - JOUR AB - Background: Risk factors for malignant tumours of the central nervous system (CNS) are largely unknown. Methods: We pooled six European cohorts (N = 302,493) and assessed the association between residential exposure to nitrogen dioxide (NO2), fine particles (PM2.5), black carbon (BC), ozone (O3) and eight elemental components of PM2.5 (copper, iron, potassium, nickel, sulfur, silicon, vanadium, and zinc) and malignant intracranial CNS tumours defined according to the International Classification of Diseases ICD-9/ICD-10 codes 192.1/C70.0, 191.0–191.9/C71.0–C71.9, 192.0/C72.2–C72.5. We applied Cox proportional hazards models adjusting for potential confounders at the individual and area-level. Results: During 5,497,514 person-years of follow-up (average 18.2 years), we observed 623 malignant CNS tumours. The results of the fully adjusted linear analyses showed a hazard ratio (95% confidence interval) of 1.07 (0.95, 1.21) per 10 μg/m³ NO2, 1.17 (0.96, 1.41) per 5 μg/m³ PM2.5, 1.10 (0.97, 1.25) per 0.5 10−5m−1 BC, and 0.99 (0.84, 1.17) per 10 μg/m³ O3. Conclusions: We observed indications of an association between exposure to NO2, PM2.5, and BC and tumours of the CNS. The PM elements were not consistently associated with CNS tumour incidence. AU - Hvidtfeldt, U.A.* AU - Chen, J.* AU - Rodopoulou, S.* AU - Strak, M.* AU - de Hoogh, K.* AU - Andersen, Z.J.* AU - Bellander, T.* AU - Brandt, J.* AU - Fecht, D.* AU - Forastiere, F.* AU - Gulliver, J.* AU - Hertel, O.* AU - Hoffmann, B.* AU - Katsouyanni, K.* AU - Ketzel, M.* AU - Leander, K.* AU - Magnusson, P.K.E.* AU - Nagel, G.* AU - Pershagen, G.* AU - Rizzuto, D.* AU - Samoli, E.* AU - So, R.* AU - Stafoggia, M.* AU - Tjønneland, A.* AU - Weinmayr, G.* AU - Wolf, K. AU - Zhang, J.* AU - Zitt, E.* AU - Brunekreef, B.* AU - Hoek, G.* AU - Raaschou-Nielsen, O.* C1 - 68426 C2 - 54617 CY - Campus, 4 Crinan St, London, N1 9xw, England SP - 656-664 TI - Long-term air pollution exposure and malignant intracranial tumours of the central nervous system: A pooled analysis of six European cohorts. JO - Br. J. Cancer VL - 129 IS - 4 PB - Springernature PY - 2023 SN - 0007-0920 ER - TY - JOUR AB - Background: In many situations, the therapeutic efficacy of CAR T cells is limited due to immune suppression and poor persistence. Immunostimulatory fusion protein (IFP) constructs have been advanced as a tool to convert suppressive signals into stimulation and thus promote the persistence of T cells, but no universal IFP design has been established so far. We now took advantage of a PD-1-CD28 IFP as a clinically relevant structure to define key determinants of IFP activity. Methods: We compared different PD-1-CD28 IFP variants in a human leukemia model to assess the impact of distinctive design choices on CAR T cell performance in vitro and a xenograft mouse model. Results: We observed that IFP constructs that putatively exceed the extracellular length of PD-1 induce T-cell response without CAR target recognition, rendering them unsuitable for tumour-specific therapy. IFP variants with physiological PD-1 length ameliorated CAR T cell effector function and proliferation in response to PD-L1+ tumour cells in vitro and prolonged survival in vivo. Transmembrane or extracellular CD28 domains were found to be replaceable by corresponding PD-1 domains for in vivo efficacy. Conclusion: PD-1-CD28 IFP constructs must mimic the physiological interaction of PD-1 with PD-L1 to retain selectivity and mediate CAR-conditional therapeutic activity. AU - Lorenzini, T.* AU - Cadilha, B.L.* AU - Obeck, H.* AU - Benmebarek, M.R.* AU - Märkl, F.* AU - Michaelides, S.* AU - Strzalkowski, T.* AU - Briukhovetska, D.* AU - Müller, P.J.* AU - Nandi, S.* AU - Winter, P.* AU - Majed, L.* AU - Grünmeier, R.* AU - Seifert, M.* AU - Rausch, S.* AU - Feuchtinger, T.* AU - Endres, S. AU - Kobold, S. C1 - 68455 C2 - 54632 CY - Campus, 4 Crinan St, London, N1 9xw, England SP - 696-705 TI - Rational design of PD-1-CD28 immunostimulatory fusion proteins for CAR T cell therapy. JO - Br. J. Cancer VL - 129 IS - 4 PB - Springernature PY - 2023 SN - 0007-0920 ER - TY - JOUR AB - BACKGROUND: Fast and accurate diagnostics are key for personalised medicine. Particularly in cancer, precise diagnosis is a prerequisite for targeted therapies, which can prolong lives. In this work, we focus on the automatic identification of gastroesophageal adenocarcinoma (GEA) patients that qualify for a personalised therapy targeting epidermal growth factor receptor 2 (HER2). We present a deep-learning method for scoring microscopy images of GEA for the presence of HER2 overexpression. METHODS: Our method is based on convolutional neural networks (CNNs) trained on a rich dataset of 1602 patient samples and tested on an independent set of 307 patient samples. We additionally verified the CNN's generalisation capabilities with an independent dataset with 653 samples from a separate clinical centre. We incorporated an attention mechanism in the network architecture to identify the tissue regions, which are important for the prediction outcome. Our solution allows for direct automated detection of HER2 in immunohistochemistry-stained tissue slides without the need for manual assessment and additional costly in situ hybridisation (ISH) tests. RESULTS: We show accuracy of 0.94, precision of 0.97, and recall of 0.95. Importantly, our approach offers accurate predictions in cases that pathologists cannot resolve and that require additional ISH testing. We confirmed our findings in an independent dataset collected in a different clinical centre. The attention-based CNN exploits morphological information in microscopy images and is superior to a predictive model based on the staining intensity only. CONCLUSIONS: We demonstrate that our approach not only automates an important diagnostic process for GEA patients but also paves the way for the discovery of new morphological features that were previously unknown for GEA pathology. AU - Pisula, J.I.* AU - Datta, R.R.* AU - Valdez, L.B.* AU - Avemarg, J.R.* AU - Jung, J.O.* AU - Plum, P.* AU - Löser, H.* AU - Lohneis, P.* AU - Meuschke, M.* AU - Dos Santos, D.P.* AU - Gebauer, F.* AU - Quaas, A.* AU - Walch, A.K. AU - Bruns, C.J.* AU - Lawonn, K.* AU - Popp, F.C.* AU - Bozek, K.* C1 - 67334 C2 - 54175 CY - Campus, 4 Crinan St, London, N1 9xw, England SP - 1369-1376 TI - Predicting the HER2 status in oesophageal cancer from tissue microarrays using convolutional neural networks. JO - Br. J. Cancer VL - 128 IS - 7 PB - Springernature PY - 2023 SN - 0007-0920 ER - TY - JOUR AB - BACKGROUND: The evidence linking ambient air pollution to bladder cancer is limited and mixed. METHODS: We assessed the associations of bladder cancer incidence with residential exposure to fine particles (PM2.5), nitrogen dioxide (NO2), black carbon (BC), warm season ozone (O3) and eight PM2.5 elemental components (copper, iron, potassium, nickel, sulfur, silicon, vanadium, and zinc) in a pooled cohort (N = 302,493). Exposures were primarily assessed based on 2010 measurements and back-extrapolated to the baseline years. We applied Cox proportional hazard models adjusting for individual- and area-level potential confounders. RESULTS: During an average of 18.2 years follow-up, 967 bladder cancer cases occurred. We observed a positive though statistically non-significant association between PM2.5 and bladder cancer incidence. Hazard Ratios (HR) were 1.09 (95% confidence interval (CI): 0.93-1.27) per 5 µg/m3 for 2010 exposure and 1.06 (95% CI: 0.99-1.14) for baseline exposure. Effect estimates for NO2, BC and O3 were close to unity. A positive association was observed with PM2.5 zinc (HR 1.08; 95% CI: 1.00-1.16 per 10 ng/m3). CONCLUSIONS: We found suggestive evidence of an association between long-term PM2.5 mass exposure and bladder cancer, strengthening the evidence from the few previous studies. The association with zinc in PM2.5 suggests the importance of industrial emissions. AU - Chen, J.* AU - Rodopoulou, S.* AU - Strak, M.* AU - de Hoogh, K.* AU - Taj, T.* AU - Poulsen, A.H.* AU - Andersen, Z.J.* AU - Bellander, T.* AU - Brandt, J.* AU - Zitt, E.* AU - Fecht, D.* AU - Forastiere, F.* AU - Gulliver, J.* AU - Hertel, O.* AU - Hoffmann, B.* AU - Hvidtfeldt, U.A.* AU - Verschuren, W.M.M.* AU - Jørgensen, J.T.* AU - Katsouyanni, K.* AU - Ketzel, M.* AU - Lager, A.* AU - Leander, K.* AU - Liu, S.* AU - Ljungman, P.* AU - Severi, G.* AU - Boutron-Ruault, M.C.* AU - Magnusson, P.K.E.* AU - Nagel, G.* AU - Pershagen, G.* AU - Peters, A. AU - Rizzuto, D.* AU - van der Schouw, Y.T.* AU - Samoli, E.* AU - Sørensen, M.* AU - Stafoggia, M.* AU - Tjønneland, A.* AU - Weinmayr, G.* AU - Wolf, K. AU - Brunekreef, B.* AU - Raaschou-Nielsen, O.* AU - Hoek, G.* C1 - 64357 C2 - 52200 CY - Campus, 4 Crinan St, London, N1 9xw, England SP - 1499-1507 TI - Long-term exposure to ambient air pollution and bladder cancer incidence in a pooled European cohort: The ELAPSE project. JO - Br. J. Cancer VL - 126 IS - 10 PB - Springernature PY - 2022 SN - 0007-0920 ER - TY - JOUR AB - BACKGROUND: Head and neck squamous cell carcinoma (HNSCC) remain a substantial burden to global health. Cell-free circulating tumour DNA (ctDNA) is an emerging biomarker but has not been studied sufficiently in HNSCC. METHODS: We conducted a single-centre prospective cohort study to investigate ctDNA in patients with p16-negative HNSCC who received curative-intent primary surgical treatment. Whole-exome sequencing was performed on formalin-fixed paraffin-embedded (FFPE) tumour tissue. We utilised RaDaRTM, a highly sensitive personalised assay using deep sequencing for tumour-specific variants, to analyse serial pre- and post-operative plasma samples for evidence of minimal residual disease and recurrence. RESULTS: In 17 patients analysed, personalised panels were designed to detect 34 to 52 somatic variants. Data show ctDNA detection in baseline samples taken prior to surgery in 17 of 17 patients. In post-surgery samples, ctDNA could be detected at levels as low as 0.0006% variant allele frequency. In all cases with clinical recurrence to date, ctDNA was detected prior to progression, with lead times ranging from 108 to 253 days. CONCLUSIONS: This study illustrates the potential of ctDNA as a biomarker for detecting minimal residual disease and recurrence in HNSCC and demonstrates the feasibility of personalised ctDNA assays for the detection of disease prior to clinical recurrence. AU - Flach, S.* AU - Howarth, K.* AU - Hackinger, S.* AU - Pipinikas, C.* AU - Ellis, P.* AU - McLay, K.* AU - Marsico, G.* AU - Forshew, T.* AU - Walz, C.* AU - Reichel, C.A.* AU - Gires, O. AU - Canis, M. AU - Baumeister, P. C1 - 64240 C2 - 52118 CY - Campus, 4 Crinan St, London, N1 9xw, England SP - 1186–1195 TI - Liquid BIOpsy for MiNimal RESidual DiSease Detection in Head and Neck Squamous Cell Carcinoma (LIONESS)-a personalised circulating tumour DNA analysis in head and neck squamous cell carcinoma. JO - Br. J. Cancer VL - 126 PB - Springernature PY - 2022 SN - 0007-0920 ER - TY - JOUR AB - BACKGROUND: CA125 is the best available yet insufficiently sensitive biomarker for early detection of ovarian cancer. There is a need to identify novel biomarkers, which individually or in combination with CA125 can achieve adequate sensitivity and specificity for the detection of earlier-stage ovarian cancer. METHODS: In the European Prospective Investigation into Cancer and Nutrition (EPIC) cohort, we measured serum levels of 92 preselected proteins for 91 women who had blood sampled ≤18 months prior to ovarian cancer diagnosis, and 182 matched controls. We evaluated the discriminatory performance of the proteins as potential early diagnostic biomarkers of ovarian cancer. RESULTS: Nine of the 92 markers; CA125, HE4, FOLR1, KLK11, WISP1, MDK, CXCL13, MSLN and ADAM8 showed an area under the ROC curve (AUC) of ≥0.70 for discriminating between women diagnosed with ovarian cancer and women who remained cancer-free. All, except ADAM8, had shown at least equal discrimination in previous case-control comparisons. The discrimination of the biomarkers, however, was low for the lag-time of >9-18 months and paired combinations of CA125 with any of the 8 markers did not improve discrimination compared to CA125 alone. CONCLUSION: Using pre-diagnostic serum samples, this study identified markers with good discrimination for the lag-time of 0-9 months. However, the discrimination was low in blood samples collected more than 9 months prior to diagnosis, and none of the markers showed major improvement in discrimination when added to CA125. AU - Mukama, T.* AU - Fortner, R.T.* AU - Katzke, V.* AU - Hynes, L.C.* AU - Petrera, A. AU - Hauck, S.M. AU - Johnson, T.* AU - Schulze, M.* AU - Schiborn, C.* AU - Rostgaard-Hansen, A.L.* AU - Tjønneland, A.* AU - Overvad, K.* AU - Pérez, M.J.S.* AU - Crous-Bou, M.* AU - Chirlaque, M.D.* AU - Amiano, P.* AU - Ardanaz, E.* AU - Watts, E.L.* AU - Travis, R.C.* AU - Sacerdote, C.* AU - Grioni, S.* AU - Masala, G.* AU - Signoriello, S.* AU - Tumino, R.* AU - Gram, I.T.* AU - Sandanger, T.M.* AU - Sartor, H.* AU - Lundin, E.* AU - Idahl, A.* AU - Heath, A.K.* AU - Dossus, L.* AU - Weiderpass, E.* AU - Kaaks, R.* C1 - 64058 C2 - 52051 CY - Campus, 4 Crinan St, London, N1 9xw, England TI - Prospective evaluation of 92 serum protein biomarkers for early detection of ovarian cancer. JO - Br. J. Cancer PB - Springernature PY - 2022 SN - 0007-0920 ER - TY - JOUR AB - BACKGROUND: Chimeric antigen receptor (CAR) T cell therapy has been successfully translated to clinical practice for the treatment of B cell malignancies. The suppressive microenvironment of many malignancies is a bottleneck preventing treatment success of CAR T cells in a broader range of tumours. Among others, the immunosuppressive metabolite adenosine is present in high concentrations within many tumours and dampens anti-tumour function of immune cells and consequently therapeutic response. METHODS: Here, we present the impact of the selective adenosine A2A and A2B receptor antagonist AB928/etrumadenant on CAR T cell cytokine secretion, proliferation, and cytotoxicity. Using phosphorylation-specific flow cytometry, we evaluated the capability of AB928 to shield CAR T cells from adenosine-mediated signalling. The effect of orally administered AB928 on CAR T cells was assessed in a syngeneic mouse model of colon carcinoma. RESULTS: We found that immunosuppressive signalling in CAR T cells in response to adenosine was fully blocked by the small molecule inhibitor. AB928 treatment enhanced CAR T cell cytokine secretion and proliferation, granted efficient cytolysis of tumour cells in vitro and augmented CAR T cell activation in vivo. CONCLUSIONS: Together our results suggest that combination therapy with AB928 represents a promising approach to improve adoptive cell therapy. AU - Seifert, M.* AU - Benmebarek, M.R.* AU - Briukhovetska, D.* AU - Märkl, F.* AU - Dörr, J.* AU - Cadilha, B.L.* AU - Jobst, J.* AU - Stock, S.* AU - Andreu-Sanz, D.* AU - Lorenzini, T.* AU - Grünmeier, R.* AU - Oner, A.* AU - Obeck, H.* AU - Majed, L.* AU - Dhoqina, D.* AU - Feinendegen, M.* AU - Gottschlich, A.* AU - Zhang, J.* AU - Schindler, U.* AU - Endres, S.* AU - Kobold, S. C1 - 66472 C2 - 52831 CY - Campus, 4 Crinan St, London, N1 9xw, England SP - 2175-2185 TI - Impact of the selective A2AR and A2BR dual antagonist AB928/etrumadenant on CAR T cell function. JO - Br. J. Cancer VL - 127 IS - 12 PB - Springernature PY - 2022 SN - 0007-0920 ER - TY - JOUR AB - Background Pre-operative treatment planning in head and neck squamous cell carcinoma (HNSCC) is mainly dictated by clinical staging, which has major shortcomings. Histologic grading is irrelevant due to its lack of prognostic impact. Recently, a novel grading termed Cellular Dissociation Grade (CDG) based on Tumour Budding and Cell Nest Size was shown to be highly prognostic for resected HNSCC. We aimed to probe the predictive and prognostic impact of CDG in the pre-operative biopsies of HNSCC. Methods We evaluated CDG in n = 160 pre-therapeutic biopsies from patients who received standardised treatment following German guidelines, and correlated the results with pre- and post-therapeutic staging data and clinical outcome. Results Pre-operative CDG was highly predictive of post-operative tumour stage, including the prediction of occult lymph node metastasis. Uni- and multivariate analysis revealed CDG to be an independent prognosticator of overall, disease-specific and disease-free survival (p < 0.001). Hazard ratio for disease-specific survival was 6.1 (11.1) for nG2 (nG3) compared with nG1 tumours. Conclusions CDG is a strong outcome predictor in the pre-treatment scenario of HNSCC and identifies patients with nodal-negative disease. CDG is a purely histology-based prognosticator in the pre-therapeutic setting that supplements clinical staging and may aide therapeutic stratification of HNSCC patients. AU - Jesinghaus, M.* AU - Steiger, K.* AU - Stögbauer, F.* AU - Haller, B.* AU - Kolk, A.* AU - Straßen, U.* AU - Pickhard, A.* AU - Wirth, M.* AU - Silva, M.* AU - Budczies, J.* AU - Becker von Rose, A.* AU - Konukiewitz, B.* AU - Kuhn, P.* AU - Klinghammer, K.* AU - Dapper, H.* AU - Münch, S.* AU - Combs, S.E. AU - Weichert, W.* AU - Boxberg, M.* C1 - 57837 C2 - 48129 CY - Macmillan Building, 4 Crinan St, London N1 9xw, England SP - 835-846 TI - Pre-operative cellular dissociation grading in biopsies is highly predictive of post-operative tumour stage and patient outcome in head and neck squamous cell carcinoma. JO - Br. J. Cancer VL - 122 IS - 6 PB - Nature Publishing Group PY - 2020 SN - 0007-0920 ER - TY - JOUR AB - BACKGROUND: Cellular Dissociation Grade (CDG) composed of tumour budding and cell nest size has been shown to independently predict prognosis in pre-therapeutic biopsies and primary resections of oesophageal squamous cell carcinoma (ESCC). Here, we aimed to evaluate the prognostic impact of CDG in ESCC after neoadjuvant therapy.METHODS: We evaluated cell nest size and tumour budding activity in 122 post-neoadjuvant ESCC resections, correlated the results with tumour regression groups and patient survival and compared the results with data from primary resected cases as well as pre-therapeutic biopsies.RESULTS: CDG remained stable when results from pre-therapeutic biopsies and post-therapeutic resections from the same patient were compared. CDG was associated with therapy response and a strong predictor of overall, disease-specific (DSS) and disease-free (DFS) survival in univariate analysis and-besides metastasis-remained the only significant survival predictor for DSS and DFS in multivariate analysis. Multivariate DFS hazard ratios reached 3.3 for CDG-G2 and 4.9 for CDG-G3 neoplasms compared with CDG-G1 carcinomas (p = 0.016).CONCLUSIONS: CDG is the only morphology-based grading algorithm published to date, which in concert with regression grading, is able to contribute relevant prognostic information in the post-neoadjuvant setting of ESCC. AU - Jesinghaus, M.* AU - Boxberg, M.* AU - Wilhelm, D.* AU - Münch, S.* AU - Dapper, H.* AU - Quante, M.* AU - Schlag, C.* AU - Lange, S.* AU - Budczies, J.* AU - Konukiewitz, B.* AU - Mollenhauer, M.* AU - Schlitter, A.M.* AU - Becker, K.F.* AU - Feith, M.* AU - Friess, H.* AU - Steiger, K.* AU - Combs, S.E. AU - Weichert, W.* C1 - 57269 C2 - 47676 CY - Macmillan Building, 4 Crinan St, London N1 9xw, England SP - 1050-1057 TI - Post-neoadjuvant cellular dissociation grading based on tumour budding and cell nest size is associated with therapy response and survival in oesophageal squamous cell carcinoma. JO - Br. J. Cancer VL - 121 IS - 12 PB - Nature Publishing Group PY - 2019 SN - 0007-0920 ER - TY - JOUR AB - BACKGROUND: The prediction of therapy response in head and neck squamous cell cancer (HNSCC) requires biomarkers, which are also a prerequisite for personalised therapy concepts. The current study aimed to identify therapy-responsive microRNAs (miRNAs) in the circulation that can serve as minimally invasive prognostic markers for HNSCC patients undergoing radiotherapy. METHODS: We screened plasma miRNAs in a discovery cohort of HNSCC patients before therapy and after treatment. We further compared the plasma miRNAs of the patients to age- and sex-matched healthy controls. All miRNAs identified as biomarker candidates were then confirmed in an independent validation cohort of HNSCC patients and tested for correlation with the clinical outcome. RESULTS: We identified a signature of eight plasma miRNAs that differentiated significantly (P=0.003) between HNSCC patients and healthy donors. MiR-186-5p demonstrated the highest sensitivity and specificity to classify HNSCC patients and healthy individuals. All therapy-responsive and patient-specific miRNAs in plasma were also detectable in tumour tissues derived from the same patients. High expression of miR-142-3p, miR-186-5p, miR-195-5p, miR-374b-5p and miR-574-3p in the plasma correlated with worse prognosis. CONCLUSIONS: Circulating miR-142-3p, miR-186-5p, miR-195-5p, miR-374b-5p and miR-574-3p represent the most promising markers for prognosis and therapy monitoring in the plasma of HNSCC patients. We found strong evidence that the circulating therapy-responsive miRNAs are tumour related and were able to validate them in an independent cohort of HNSCC patients. AU - Summerer, I. AU - Unger, K. AU - Braselmann, H. AU - Schuettrumpf, L. AU - Maihoefer, C. AU - Baumeister, P. AU - Kirchner, T.* AU - Niyazi, M. AU - Sage, E.H.* AU - Specht, H.M.* AU - Multhoff, G. AU - Mörtl, S. AU - Belka, C. AU - Zitzelsberger, H. C1 - 45131 C2 - 37245 CY - London SP - 76-82 TI - Circulating microRNAs as prognostic therapy biomarkers in head and neck cancer patients. JO - Br. J. Cancer VL - 113 IS - 1 PB - Nature Publishing Group PY - 2015 SN - 0007-0920 ER - TY - JOUR AB - Background:Patients with UICC/AJCC stage II colon cancer have a high 5-year overall survival rate after surgery. Nevertheless, a significant subgroup of patients develops tumour recurrence. Currently, there are no clinically established biomarkers available to identify this patient group. We applied reverse-phase protein arrays (RPPA) for phosphatidylinositide-3-kinase pathway activation mapping to stratify patients according to their risk of tumour recurrence after surgery.Methods:Full-length proteins were extracted from formalin-fixed, paraffin-embedded tissue samples of 118 patients who underwent curative resection. RPPA technology was used to analyse expression and/or phosphorylation levels of six major factors of the phosphatidylinositide-3-kinase pathway. Oncogenic mutations of KRAS and BRAF, and DNA microsatellite status, currently discussed as prognostic markers, were analysed in parallel.Results:Expression of phospho-AKT (HR=3.52; P=0.032), S6RP (HR=6.3; P=0.044), and phospho-4E-BP1 (HR=4.12; P=0.011) were prognostic factors for disease-free survival. None of the molecular genetic alterations were significantly associated with prognosis.Conclusions:Our data indicate that activation of the PI3K/AKT pathway evidenced on the protein level might be a valuable prognostic marker to stratify patients for their risk of tumour recurrence. Beside adjuvant chemotherapy targeting of upregulated PI3K/AKT signalling may be an attractive strategy for treatment of high-risk patients. AU - Malinowsky, K.* AU - Nitsche, U.* AU - Janssen, K.P.* AU - Bader, F.G.* AU - Späth, C.* AU - Drecoll, E.* AU - Keller, G.* AU - Höfler, H. AU - Slotta-Huspenina, J.* AU - Becker, K.F.* C1 - 30873 C2 - 33984 CY - London SP - 2081-2089 TI - Activation of the PI3K/AKT pathway correlates with prognosis in stage II colon cancer. JO - Br. J. Cancer VL - 110 IS - 8 PB - Nature Publishing Group PY - 2014 SN - 0007-0920 ER - TY - JOUR AB - Background:The mechanisms of brain metastasis in renal cell cancer (RCC) patients are poorly understood. Chemokine and chemokine receptor expression may contribute to the predilection of RCC for brain metastasis by recruitment of monocytes/macrophages and by control or induction of vascular permeability of the blood-brain barrier.Methods:Frequency and patterns of brain metastasis were determined in 246 patients with metastatic RCC at autopsy. Expression of CXCR4, CCL7 (MCP-3), CCR2 and CD68+ tumour-associated macrophages (TAMs) were analysed in a separate series of 333 primary RCC and in 48 brain metastases using immunohistochemistry.Results:Fifteen percent of 246 patients with metastasising RCC had brain metastasis. High CXCR4 expression levels were found in primary RCC and brain metastases (85.7% and 91.7%, respectively). CCR2 (52.1%) and CCL7 expression (75%) in cancer cells of brain metastases was more frequent compared with primary tumours (15.5% and 16.7%, respectively; P<0.0001 each). The density of CD68+ TAMs was similar in primary RCC and brain metastases. However, TAMs were more frequently CCR2-positive in brain metastases than in primary RCC (P<0.001).Conclusion:Our data demonstrate that the monocyte-specific chemokine CCL7 and its receptor CCR2 are expressed in tumour cells of RCC. We conclude that monocyte recruitment by CCR2 contributes to brain metastasis of RCC. AU - Wyler, L.* AU - Napoli, C.U.* AU - Ingold, B.* AU - Sulser, T.* AU - Heikenwälder, M. AU - Schraml, P.* AU - Moch, H.* C1 - 29053 C2 - 33605 CY - London SP - 686-694 TI - Brain metastasis in renal cancer patients: Metastatic pattern, tumour-associated macrophages and chemokine/chemoreceptor expression. JO - Br. J. Cancer VL - 110 IS - 3 PB - Nature Publishing Group PY - 2014 SN - 0007-0920 ER - TY - JOUR AB - Background: Most of the heritable risk of glioma is presently unaccounted for by mutations in known genes. In addition to rare inactivating germline mutations in TP53 causing glioma in the context of the Li-Fraumeni syndrome, polymorphic variation in TP53 may also contribute to the risk of developing glioma. Methods: To comprehensively evaluate the impact of variation in TP53 on risk, we analysed 23 tagSNPs and imputed 2377 unobserved genotypes in four series totaling 4147 glioma cases and 7435 controls. Results: The strongest validated association signal was shown by the imputed single-nucleotide polymorphism (SNP) rs78378222 (P = 6.86 x 10(-24), minor allele frequency similar to 0.013). Confirmatory genotyping confirmed the high quality of the imputation. The association between rs78378222 and risk was seen for both glioblastoma multiforme (GBM) and non- GBM tumours. We comprehensively examined the relationship between rs78378222 and overall survival in two of the case series totaling 1699 individuals. Despite employing statistical tests sensitive to the detection of differences in early survival, no association was shown. Conclusion: Our data provided strong validation of rs78378222 as a risk factor for glioma but do not support the tenet that the polymorphism being a clinically useful prognostic marker. Acquired TP53 inactivation is a common feature of glioma. As rs78378222 changes the polyadenylation signal of TP53 leading to impaired 3'-end processing of TP53 mRNA, the SNP has strong plausibility for being directly functional contributing to the aetiological basis of glioma. AU - Enciso-Mora, V.* AU - Hosking, F.J.* AU - di Stefano, A.L.* AU - Zelenika, D.* AU - Shete, S.* AU - Broderick, P.* AU - Idbaih, A.* AU - Delattre, J.Y.* AU - Hoang-Xuan, K.* AU - Marie, Y.* AU - Labussière, M.* AU - Alentorn, A.* AU - Ciccarino, P.* AU - Rossetto, M.* AU - Armstrong, G.* AU - Liu, Y.* AU - Gousias, K.* AU - Schramm, J.* AU - Lau, C.* AU - Hepworth, S.J.* AU - Schoemaker, M.* AU - Strauch, K. AU - Müller-Nurasyid, M. AU - Schreiber, S.* AU - Franke, A.* AU - Moebus, S.* AU - Eisele, L.* AU - Swerdlow, A.* AU - Simon, M.* AU - Bondy, M.* AU - Lathrop, M* AU - Sanson, M.* AU - Houlston, R.S.* C1 - 25513 C2 - 31873 SP - 2178-2185 TI - Low penetrance susceptibility to glioma is caused by the TP53 variant rs78378222. JO - Br. J. Cancer VL - 108 IS - 10 PB - Nature Publishing PY - 2013 SN - 0007-0920 ER - TY - JOUR AB - Background:MiR-221/-222 are frequently overexpressed in breast cancer and are associated with increased malignancy. The specific modification of microRNAs (miRNAs) expression could be a promising strategy in breast cancer therapy, leading to the suppression of tumourigenic processes in tumour cells.Methods:MiR-221/-222 expressions were analysed in 86 breast cancer tissues by quantitative RT-PCR and tested for correlation with immunohistochemistry data and clinical follow-up. In vitro assays were conducted using human breast cancer cell lines with lentiviral overexpression of miR-221/-222.Results:In tumour tissues, miR-221/-222 were associated with the occurrence of distant metastases. In particular, high levels of miR-221 were revealed to have a high prognostic impact for the identification of significantly different groups with advanced tumours. MiR-221/-222 overexpression strongly increased cell proliferation and invasion in vitro. Following miR-221/-222 overexpression an increased uPAR expression and cell invasion were observed.Conclusion:This study demonstrates a significant role for highly expressed miR-221/-222 in advanced breast cancers allowing for the identification of significantly different prognostic groups, particularly for HER2-positive and lymph-node-positive breast cancers. Considering that miR-221/-222 are strongly involved in cell invasion, these miRNAs may be promising markers for breast cancer prognosis and therapy. AU - Falkenberg, N. AU - Anastasov, N. AU - Rappl, K. AU - Braselmann, H. AU - Auer, G.* AU - Walch, A.K. AU - Huber, M. AU - Höfig, I. AU - Schmitt, M.* AU - Höfler, H. AU - Atkinson, M.J. AU - Aubele, M. C1 - 27951 C2 - 32874 SP - 2714-2723 TI - MiR-221/-222 differentiate prognostic groups in advanced breast cancers and influence cell invasion. JO - Br. J. Cancer VL - 109 IS - 10 PB - Nature Publishing PY - 2013 SN - 0007-0920 ER - TY - JOUR AB - Background:Genomic rearrangements at the fragile site FRA1E may disrupt the dihydropyrimidine dehydrogenase gene (DPYD) gene which is involved in 5-fluorouracil (5-FU) catabolism. In triple-negative breast cancer (TNBC), a subtype of breast cancer frequently deficient in DNA repair, we have investigated the susceptibility to acquire copy number variations (CNVs) in DPYD and evaluated their impact on standard adjuvant treatment.Methods:DPYD CNVs were analysed in 106 TNBC tumour specimens using multiplex ligation-dependent probe amplification (MLPA) analysis. Dihydropyrimidine dehydrogenase (DPD) expression was determined by immunohistochemistry in 146 tumour tissues.Results:In TNBC, we detected 43 (41%) tumour specimens with genomic deletions and/or duplications within DPYD which were associated with higher histological grade (P=0.006) and with rearrangements in the DNA repair gene BRCA1 (P=0.007). Immunohistochemical analysis revealed low, moderate and high DPD expression in 64%, 29% and 7% of all TNBCs, and in 40%, 53% and 7% of TNBCs with DPYD CNVs, respectively. Irrespective of DPD protein levels, the presence of CNVs was significantly related to longer time to progression in patients who had received 5-FU- and/or anthracycline-based polychemotherapy (hazard ratio=0.26 (95% CI: 0.07-0.91), log-rank P=0.023; adjusted for tumour stage: P=0.037).Conclusion:Genomic rearrangements in DPYD, rather than aberrant DPD protein levels, reflect a distinct tumour profile associated with prolonged time to progression upon first-line chemotherapy in TNBC. AU - Gross, E.* AU - Meul, C.* AU - Raab, S.* AU - Propping, C.* AU - Avril, S.* AU - Aubele, M. AU - Gkazepis, A.* AU - Schuster, T.* AU - Grebenchtchikov, N.* AU - Schmitt, M.* AU - Kiechle, M.* AU - Meijer, J.* AU - Vijzelaar, R.* AU - Meindl, A.* AU - van Kuilenburg, A.B.* C1 - 27927 C2 - 32862 SP - 2347-2355 TI - Somatic copy number changes in DPYD are associated with lower risk of recurrence in triple-negative breast cancers. JO - Br. J. Cancer VL - 109 IS - 9 PB - Nature Publishing PY - 2013 SN - 0007-0920 ER - TY - JOUR AB - Background: Smoking is not associated with prostate cancer incidence in most studies, but associations between smoking and fatal prostate cancer have been reported. Methods: During 1992 and 2000, lifestyle information was assessed via questionnaires and personal interview in a cohort of 145112 European men. Until 2009, 4623 incident cases of prostate cancer were identified, including 1517 cases of low-grade, 396 cases of high grade, 1516 cases of localised, 808 cases of advanced disease, and 432 fatal cases. Multivariable Cox proportional hazards regression models were used to examine the association of smoking status, smoking intensity, and smoking duration with the risk of incident and fatal prostate cancer. Results: Compared with never smokers, current smokers had a reduced risk of prostate cancer (RR = 0.90, 95% CI: 0.83-0.97), which was statistically significant for localised and low-grade disease, but not for advanced or high-grade disease. In contrast, heavy smokers (25+ cigarettes per day) and men who had smoked for a long time (40+ years) had a higher risk of prostate cancer death (RR = 1.81, 95% CI: 1.11-2.93; RR = 1.38, 95% CI: 1.01-1.87, respectively). Conclusion: The observation of an increased prostate cancer mortality among heavy smokers confirms the results of previous prospective studies. AU - Rohrmann, S.* AU - Linseisen, J. AU - Allen, N.* AU - Bueno-de-Mesquita, H.B.* AU - Johnsen, N.F.* AU - Tjonneland, A.* AU - Overvad, K.* AU - Kaaks, R.* AU - Teucher, B.* AU - Boeing, H.* AU - Pischon, T.* AU - Lagiou, P.* AU - Trichopoulou, A.* AU - Trichopoulos, D.* AU - Palli, D.* AU - Krogh, V.* AU - Tunnino, R.* AU - Ricceri, F.* AU - Suarez, M.V.A.* AU - Agudo, A.* AU - Sánchez, M.J.* AU - Chirlaque, M.D.* AU - Barricarte, A.* AU - Larrañaga, N.* AU - Boshuizen, H.* AU - van Kranen, H.J.* AU - Stettin, P.* AU - Johansson, M.* AU - Bjartell, A.* AU - Ulmert, D.* AU - Khaw, K.T.* AU - Wareham, N.J.* AU - Ferrari, P.* AU - Romieux, I.* AU - Gunter, M.J.R.* AU - Riboli, E.* AU - Key, T.J.* C1 - 24112 C2 - 31325 SP - 708-714 TI - Smoking and the risk of prostate cancer in the European Prospective Investigation into Cancer and Nutrition. JO - Br. J. Cancer VL - 108 IS - 3 PB - Nature Publishing Group PY - 2013 SN - 0007-0920 ER - TY - JOUR AB - Background:Oesophageal adenocarcinomas often show resistances to chemotherapy (CTX), therefore, it would be of high interest to better understand the mechanisms of resistance. We examined the expression of heat-shock proteins (HSPs) and glucose-regulated proteins (GRPs) in pretherapeutic biopsies of oesophageal adenocarcinomas to assess their potential role in CTX response.Methods:Ninety biopsies of locally advanced adenocarcinomas before platin/5-fluorouracil (FU)-based CTX were investigated by reverse phase protein arrays (RPPAs), immunohistochemistry (IHC) and quantitative RT-PCR.Results:CTX response strongly correlated with survival (P=0.001). Two groups of tumours with specific protein expression patterns were identified by RPPA: Group A was characterised by low expression of HSP90, HSP27 and p-HSP27((Ser15, Ser78, Ser82)) and high expression of GRP78, GRP94, HSP70 and HSP60; Group B exhibited the inverse pattern. Tumours of Group A were more likely to respond to CTX, resulting in histopathological tumour regression (P=0.041) and post-therapeutic down-categorisation from cT3 to ypT0-T2 (P=0.040). High HSP60 protein (IHC) and mRNA expression were also associated with tumour down-categorisation (P=0.016 and P=0.004).Conclusion:Our findings may enhance the understanding of CTX response mechanisms, might be helpful to predict CTX response and might have translational relevance as they highlight the role of potentially targetable cellular stress proteins in the context of CTX response. AU - Slotta-Huspenina, J.* AU - Wolff, C.* AU - Drecoll, E.* AU - Feith, M.* AU - Bettstetter, M.* AU - Malinowsky, K.* AU - Bauer, L.* AU - Becker, K.* AU - Ott, K.* AU - Höfler, H. AU - Becker, K.F.* AU - Langer, R.* C1 - 26242 C2 - 32139 SP - 370-378 TI - A specific expression profile of heat-shock proteins and glucose-regulated proteins is associated with response to neoadjuvant chemotherapy in oesophageal adenocarcinomas. JO - Br. J. Cancer VL - 109 IS - 2 PB - Nature Publishing PY - 2013 SN - 0007-0920 ER - TY - JOUR AB - BACKGROUND: Lignans - oestrogenic substances present in various foods - are associated with postmenopausal breast cancer risk, but not much is known regarding their effects on survival. METHODS: In a follow-up study of 2653 postmenopausal breast cancer patients diagnosed between 2001 and 2005, vital status and causes of death were verified through end of 2009. Hazard ratios (HRs) and 95% confidence intervals (CIs) for estimated enterolignans, lignan-rich foods, and dietary fibre in relation to overall survival (OS) and breast cancer-specific survival (BCSS) were assessed using Cox proportional hazards models stratified by age at diagnosis and adjusted for prognostic/confounding factors. RESULTS: Median follow-up time was 6.4 years, and 321 women died, 235 with breast cancer. High estimated enterolactone and enterodiol levels were associated with significantly lower overall mortality (highest quintile, HR=0.60, 95% CI=0.40-0.89, P(Trend)=0.02 and HR=0.63, 95% CI=0.42-0.95, P(Trend)=0.02, respectively). Fibre intake was also associated with a significantly lower overall mortality. Differentiated by median fibre intake, associations with estimated enterolignans were still evident at low but not high fibre intake. There was no effect modification by oestrogen receptor status and menopausal hormone therapy. CONCLUSION: Postmenopausal breast cancer patients with high estimated enterolignans may have a better survival. AU - Buck, K.* AU - Zaineddin, A.K.* AU - Vrieling, A.* AU - Heinz, J.* AU - Linseisen, J. AU - Flesch-Janys, D.* AU - Chang-Claude, J.* C1 - 6884 C2 - 29394 SP - 1151-1157 TI - Estimated enterolignans, lignan-rich foods, and fibre in relation to survival after postmenopausal breast cancer. JO - Br. J. Cancer VL - 105 IS - 8 PB - Nature Publishing Group PY - 2011 SN - 0007-0920 ER - TY - JOUR AB - BACKGROUND: Protein tyrosine kinase 6 (PTK6; breast tumour kinase) is overexpressed in up to 86% of the invasive breast cancers, and its association with the oncoprotein human epidermal growth factor receptor 2 (HER2) was shown in vitro by co-precipitation. Furthermore, expression of PTK6 in tumours is linked with the expression of HER2. METHOD AND RESULTS: In this study, we used the proximity ligation assay (PLA) technique on formalin-fixed paraffin sections from eighty invasive breast carcinoma tissue specimens to locate PTK6-HER2 protein-protein complexes. Proximity ligation assay signals from protein complexes were assessed quantitatively, and expression levels showed a statistically significant association with tumour size (P=0.015) and course of the cancer disease (P=0.012). CONCLUSION: Protein tyrosine kinase 6 forms protein complexes with HER2 in primary breast cancer tissues, which can be visualised by use of the PLA technique. Human epidermal growth factor receptor 2-PTK6 complexes are of prognostic relevance. AU - Aubele, M. AU - Spears, M.* AU - Ludyga, N. AU - Braselmann, H. AU - Feuchtinger, A. AU - Taylor, K.J.* AU - Lindner, K. AU - Auer, G.* AU - Stering, K. AU - Höfler, H. AU - Schmitt, M.* AU - Bartlett, J.M.S.* C1 - 5191 C2 - 27398 SP - 663-667 TI - In situ quantification of HER2-protein tyrosine kinase 6 (PTK6) protein-protein complexes in paraffin sections from breast cancer tissues. JO - Br. J. Cancer VL - 103 IS - 5 PB - Nature Publ. Group PY - 2010 SN - 0007-0920 ER - TY - JOUR AB - BACKGROUND: Poor reporting compromises the reliability and clinical value of prognostic tumour marker studies. We review articles to assess the reporting of patients and events using REMARK guidelines, at the time of guideline publication. METHODS: We sampled 50 prognostic tumour marker studies from higher impact cancer journals between 2006 and 2007. The inclusion criteria were cancer; focus on single biological tumour marker; survival analysis; multivariable analysis; and not gene array or proteomic data. Articles were assessed for the REMARK profile and other REMARK guideline items. We propose a reporting aid, the REMARK profile, motivated by the CONSORT flowchart. RESULTS: In 50 studies assessed for the REMARK profile, the number of eligible patients (56% of articles), excluded patients (54%) and patients in analyses (98%) was reported. Only 50% of articles reported the number of outcome events. In multivariable analyses, 54% and 30% of articles reported patient and event numbers for all variables. Of the studies, 66% used archival samples, indicating a potentially biased patient selection. Only 36% of studies reported clearly defined outcomes. CONCLUSIONS: Good reporting is critical for the interpretability and clinical applicability of prognostic studies. Current reporting of key information, such as the number of outcome events in all patients and subgroups, is poor. Use of the REMARK profile would greatly improve reporting and enhance prognostic research. AU - Mallett, S.* AU - Timmer, A. AU - Sauerbrei, W.* AU - Altman, D.G.* C1 - 929 C2 - 27141 SP - 173-180 TI - Reporting of prognostic studies of tumour markers: A review of published articles in relation to REMARK guidelines. JO - Br. J. Cancer VL - 102 IS - 1 PB - Nature Publ. Group PY - 2010 SN - 0007-0920 ER - TY - JOUR AB - Ras acts in signalling pathways regulating the activity of multiple cellular functions including cell proliferation, differentiation, and apoptosis. Amino-acid exchanges at position 12, 13, or 61 of the Kras gene convert the proto-oncogene into an activated oncogene. Until now, a direct comparison of genome-wide expression profiling studies of Kras overexpression and different Kras mutant forms in a single assay system has not been carried out. In our study, we focused on the direct comparison of global gene expression effects caused by mutations in codon 12 or 13 of the Kras gene and Kras overexpression in murine fibroblasts. We determined Kras cellular mRNA, Ras protein and activated Ras protein levels. Further, we compared our data to the proteome analysis of the same transfected cell lines. Both overexpression and mutations of Kras lead to common altered gene expression patterns. Only two genes, Lox and Col1a1, were reversely regulated in the Kras transfectants. They may contribute to the higher aggressiveness of the Kras codon 12 mutation in tumour progression. The functional annotation of differentially expressed genes revealed a high frequency of proteins involved in tumour growth and angiogenesis. These data further support the important role of these genes in tumour-associated angiogenesis. AU - Horsch, M. AU - Recktenwald, C.V.* AU - Schädler, S. AU - Hrabě de Angelis, M. AU - Seliger, B.* AU - Beckers, J. C1 - 691 C2 - 26200 SP - 656-662 TI - Overexpressed vs mutated Kras in murine fibroblasts: A molecular phenotyping study. JO - Br. J. Cancer VL - 100 IS - 4 PB - Nature Publ. Group PY - 2009 SN - 0007-0920 ER - TY - JOUR AB - Background: The aim of this study was to investigate the prognostic effect of tumour-infiltrating lymphocytes (TILs) in serous stage III ovarian carcinoma to determine TIL clonality and to correlate this to Her2/neu expression. Methods: Formalin-fixed and paraffin-embedded ovarian carcinomas were examined for CD20-, CD3-, CD4- and CD8-positive lymphocytes (n=100), and for Her2/neu-positive tumour cells (n=55/100) by immunohistochemistry. Clonality analysis was carried out by T-cell receptor italic gamma (TCRitalic gamma) gene rearrangements (n=93/100). Statistical analyses included experimental and clinico-pathological variables, as well as disease-free (DFS) and overall (OS) survival. Results: CD20-positive B lymphocytes were present in 57.7% (stromal)/33.0% (intraepithelial) and CD3-positive T lymphocytes in 99.0% (stromal)/90.2% (intraepithelial) of ovarian carcinomas. Intraepithelial CD3-positive T lymphocytes were correlated with improved DFS in optimally debulked patients (P=0.0402). Intraepithelial CD8-positive T lymphocytes were correlated with improved OS in all optimally debulked patients (P=0.0201) and in those undergoing paclitaxel/carboplatin therapy (P=0.0092). Finally, rarified and clonal TCRitalic gamma gene rearrangements were detected in 37 out of 93 (39.8%) and 15 out of 93 (16.1%) cases, respectively. This was marginally associated with improved DFS (P=0.0873). Despite a significant correlation of HER2/neu status and intraepithelial CD8-positive lymphocytes (P=0.0264), this was non-directional (R=-0.257; P=0.0626). Conclusion: Improved survival of ovarian cancer patients is related to the infiltration, clonal selection and intraepithelial persistence of T lymphocytes. AU - Stumpf, M.* AU - Hasenburg, A.* AU - Riener, M.O.* AU - Jütting, U. AU - Wang, C.* AU - Shen, Y.* AU - Orlowska-Volk, M.* AU - Fisch, P.* AU - Wang, Z.* AU - Gitsch, G.* AU - Werner, M.* AU - Lassmann, S.* C1 - 814 C2 - 26405 CY - London SP - 1513-1521 TI - Intraepithelial CD8-positive T lymphocytes predict survival for patients with serous stage III ovarian carcinomas: Relevance of clonal selection of T lymphocytes. JO - Br. J. Cancer VL - 101 IS - 9 PB - Nature Publ. Group PY - 2009 SN - 0007-0920 ER - TY - JOUR AB - The cytoplasmic tyrosine kinase PTK6 (BRK) shows elevated expression in approximately two-thirds of primary breast tumours, and is implicated in EGF receptor-dependent signalling and epithelial tumorigenesis. Using immunohistochemistry, we performed a retrospective study on 426 archival breast cancer samples from patients with long-term follow-up and compared the protein expression levels of PTK6, the HER receptors, Sam68 (a substrate of PTK6), and signalling proteins including MAP kinase (MAPK), phosphorylated MAPK (P-MAPK), and PTEN. We show that PTK6 expression is of significant prognostic value in the outcome of breast carcinomas. In multivariate analysis, the disease-free survival of patients of >or=240 months was directly associated with the protein expression level of PTK6 (P AU - Aubele, M. AU - Walch, A.K. AU - Ludyga, N. AU - Braselmann, H. AU - Atkinson, M.J. AU - Luber, B.* AU - Auer, G.* AU - Tapio, S. AU - Cooke, T.* AU - Bartlett, J.M.* C1 - 3542 C2 - 25753 SP - 1089-1095 TI - Prognostic value of protein tyrosine kinase 6 (PTK6) for long-term survival of breast cancer patients. JO - Br. J. Cancer VL - 99 IS - 7 PB - Nature Publ. Group PY - 2008 SN - 0007-0920 ER - TY - JOUR AB - Epithelial ovarian cancer is the leading cause of death among female genital malignancies. Reduced expression of the cell adhesion molecule E-cadherin was previously shown to be associated with adverse prognostic features. The role of the E-cadherin repressor Snail in ovarian cancer progression remains to be elucidated. We analysed formalin-fixed and paraffin-embedded specimens of 48 primary ovarian tumours and corresponding metastases for expression of E-cadherin and Snail by immunohistochemistry. We found a significant correlation between E-cadherin expression in primary cancers and their corresponding metastases (P<0.001). This correlation was found for Snail expression as well (P<0.001). There was a significant (P=0.008) association of reduced E-cadherin expression in primary ovarian cancer with shorter overall survival. Similarly, Snail expression in corresponding metastases (P=0.047) was associated with reduced overall survival of the patients. Additionally, the group of patients showing reduced E-cadherin and increased Snail immunoreactivity in primary tumours and corresponding metastases, respectively, had a significantly higher risk of death (P=0.002 and 0.022, respectively) when compared to the patient group with the reference expression profile E-cadherin positive and Snail negative. Taken together, the results of our study show that the E-cadherin repressor Snail is associated with lower overall survival of ovarian cancer patients. AU - Blechschmidt, K.* AU - Sassen, S.* AU - Schmalfeldt, B.* AU - Schuster, T.* AU - Höfler, H. AU - Becker, K.F.* C1 - 3103 C2 - 25202 SP - 489-495 TI - The E-cadherin repressor Snail is associated with lower overall survival of ovarian cancer patients. JO - Br. J. Cancer VL - 98 IS - 2 PB - Nature Publ. Group PY - 2008 SN - 0007-0920 ER - TY - JOUR AB - Pancreatic cancer has an abysmal prognosis. Targets for early detection, prevention and therapy are desperately needed. Inflammatory pathways have an important impact on tumour growth and progression. Expression of BLT2 (the second leukotriene B(4) receptor) was investigated by real-time RT-PCR and immunohistochemistry. Cell proliferation was studied after stable transfection with BLT2, after treatment with siRNA and Compound A as an agonist. BLT2 is expressed in all pancreatic cancer cell lines. Results from real-time RT-PCR revealed significant overexpression of BLT2 in malignant intraductal papillary mucinous neoplasias (IPMNs) and pancreatic adenocarcinoma. Intense staining was evident in IPMNs, infiltrating tumour cells and advanced pancreatic intraepithelial neoplasias (PanINs) but not in normal ductal cells. Overexpression of BLT2 as well as stimulation of Colo357, Panc-1 and AsPC1 cells with Compound A caused a significant increase in tumour cell proliferation, an effect reversed after siRNA treatment. This study demonstrates for the first time the expression of BLT2 in the pancreas and overexpression in pancreatic cancers and malignant IPMNs in particular. Upregulation of BLT2 is already evident in precursor lesions (PanINs, IPMNs). Overexpression of this receptor leads to significant growth stimulation. Therefore, we suggest BLT2 as a new target for chemoprevention and therapy for pancreatic cancer. AU - Hennig, R.* AU - Osman, T.* AU - Esposito, I. AU - Giese, N.* AU - Rao, S.M.* AU - Ding, X.Z.* AU - Tong, W.G.* AU - Büchler, M.W.* AU - Yokomizo, T.* AU - Friess, H.* AU - Adrian, T.E.* C1 - 2335 C2 - 25756 SP - 1064-1073 TI - BLT2 is expressed in PanINs, IPMNs, pancreatic cancer and stimulates tumour cell proliferation. JO - Br. J. Cancer VL - 99 IS - 7 PB - Nature Publ. Group PY - 2008 SN - 0007-0920 ER - TY - JOUR AB - Altered histone deacetylase (HDAC) activity has been identified in several types of cancer. This study was designed to determine the safety and maximum tolerated dose (MTD) of valproic acid (VPA) as an HDAC inhibitor in cancer patients. Twenty-six pre-treated patients with progressing solid tumours were enrolled in dose-escalating three-patient cohorts, starting at a dose of VPA 30 mg kg(-1) day(-1). VPA was administered as an 1-h infusion daily for 5 consecutive days in a 21-day cycle. Neurocognitive impairment dominated the toxicity profile, with grade 3 or 4 neurological side effects occurring in 8 out of 26 patients. No grade 3 or 4 haematological toxicity was observed. The MTD of infusional VPA was 60 mg kg(-1) day(-1). Biomonitoring of peripheral blood lymphocytes demonstrated the induction of histone hyperacetylation in the majority of patients and downmodulation of HDAC2. Pharmacokinetic studies showed increased mean and maximum serum VPA concentrations >120 and >250 mg l(-1), respectively, in the 90 and 120 mg kg(-1) cohorts, correlating well with the incidence of dose-limiting toxicity (DLT). Neurotoxicity was the main DLT of infusional VPA, doses up to 60 mg kg(-1) day(-1) for 5 consecutive days are well tolerated and show detectable biological activity. Further investigations are warranted to evaluate the effectivity of VPA alone and in combination with other cytotoxic drugs. AU - Atmaca, A.* AU - Al-Batran, S.E.* AU - Maurer, A.* AU - Neumann, A.* AU - Heinzel, T. AU - Hentsch, B.* AU - Schwarz, S.E.* AU - Hövelmann, S.* AU - Göttlicher, M. AU - Knuth, A.* AU - Jäger, E.* C1 - 1153 C2 - 24563 SP - 177-182 TI - Valproic acid (VPA) in patients with refractory advanced cancer: A dose escalating phase I clinical trial. JO - Br. J. Cancer VL - 97 IS - 2 PB - Nature Publ. Group PY - 2007 SN - 0007-0920 ER - TY - JOUR AB - The HER receptors are of therapeutic and prognostic significance in breast cancer, and their function is modulated by cytoplasmic tyrosine kinases like PTK6 (brk). We performed a retrospective study on archival breast cancer samples from patients with long follow-up and compared the protein expression between individual HERs and between HERs and the PTK6. Univariate and multivariate analyses were used to study the prognostic value of parameters. Metastases-free survival of patients for longer than 240 months was inversely associated (P< or =0.05) with nodal status, tumour size, and oestrogen receptor status, but was also directly associated with high protein expression levels of HER4 and PTK6 in Kaplan-Meier analysis. In multivariate analysis for metastases-free survival of >240 months, the stepwise selected parameters were tumour size (relative risk 3.1), PTK6 expression (0.4), and number of positive lymph nodes (1.2). Furthermore, we demonstrated a timedependence of the prognostic value attributed to the parameters. The HER receptors (HER2,4), but not PTK6, were independent prognostic markers for metastases-free survival at 60 months, whereas at 240 months PTK6 is the strongest prognostic marker. We demonstrate that PTK6 is a prognostic marker of metastases-free survival in breast cancer, and is independent of the classical morphological and molecular markers of lymph node involvement, tumour size, and HER2 status. AU - Aubele, M. AU - Auer, G.* AU - Walch, A.K. AU - Munro, A.* AU - Atkinson, M.J. AU - Braselmann, H. AU - Fornander, T.* AU - Bartlett, J.M.S.* C1 - 3442 C2 - 24372 SP - 801-807 TI - PTK (protein tyrosine kinase)-6 and HER2 and 4, but not HER1 and 3 predict long-term survival in breast carcinomas. JO - Br. J. Cancer VL - 96 IS - 5 PB - Nature Publ. Group PY - 2007 SN - 0007-0920 ER - TY - JOUR AB - Although epithelial cell adhesion/activating molecule (EpCAM/CD326) is one of the first tumour-associated antigens identified, it has never received the same level of attention as other target proteins for therapy of cancer. It is also striking that ever since its discovery in the late 1970s the actual contribution of EpCAM to carcinogenesis remained unexplored until very recently. With a First International Symposium on EpCAM Biology and Clinical Application this is now changing. Key topics discussed at the meeting were the frequency and level of EpCAM expression on various cancers and its prognostic potential, the role of EpCAM as an oncogenic signalling molecule for cancer cells, recent progress on EpCAM-directed immunotherapeutic approaches in clinical development and the interaction of EpCAM with other proteins, which may provide a basis for a therapeutic window and repression of its growth-promoting signalling in carcinoma. Future research on EpCAM may benefit from a unified nomenclature and more frequent exchange among those who have been working on this cancer target during the past 30 years and will do so in the future. AU - Baeuerle, P.A.* AU - Gires, O. C1 - 3455 C2 - 24657 SP - 417-423 TI - EpCAM (CD326) finding its role in cancer. JO - Br. J. Cancer VL - 96 IS - 3 PB - Nature Publ. Group PY - 2007 SN - 0007-0920 ER - TY - JOUR AB - Epithelial cell adhesion molecule EpCAM is a transmembrane glycoprotein that is frequently overexpressed in a variety of carcinomas. This pan-carcinoma antigen has served as the target for a plethora of immunotherapies. Innovative therapeutic approaches include the use of trifunctional antibodies (trAbs) that recruit and activate different types of immune effector cells at the tumour site. The trAb catumaxomab has dual specificity for EpCAM and CD3. In patients with malignant ascites, catumaxomab significantly increased the paracentesis-free interval, corroborating the high efficacy of this therapeutic antibody. Here, we characterised the monoclonal antibody (mAb) HO-3, that is, the EpCAM-binding arm of catumaxomab. Peptide mapping indicated that HO-3 recognises a discontinuous epitope, having three binding sites in the extracellular region of EpCAM. Studies with glycosylation-deficient mutants showed that mAb HO-3 recognised EpCAM independently of its glycosylation status. High-affinity binding was not only detected for mAb HO-3, but also for the monovalent EpCAM-binding arm of catumaxomab with an excellent K(D) of 5.6 x 10(-10) M. Furthermore, trAb catumaxomab was at least a 1000-fold more effective in eliciting the eradication of tumour cells by effector peripheral blood mononuclear cells compared with mAb HO-3. These findings suggest the great therapeutic potential of trAbs and clearly speak in favour of EpCAM-directed cancer immunotherapies. AU - Ruf, P.* AU - Gires, O. AU - Jäger, M.* AU - Fellinger, K. AU - Atz, J.* AU - Lindhofer, H.* C1 - 4158 C2 - 25093 SP - 315-321 TI - Characterisation of the new EpCAM-specific antibody HO-3: Implications for trifunctional antibody immunotherapy of cancer. JO - Br. J. Cancer VL - 97 IS - 3 PB - Nature Publ. Group PY - 2007 SN - 0007-0920 ER - TY - JOUR AB - Tissue samples from 13 post-Chernobyl childhood thyroid tumours that occurred within a short period of time (4-8 years) after the Chernobyl accident have been investigated by interphase FISH analysis for rearrangements of RET. In all, 77% of cases showed RET/PTC rearrangements and a distinct intratumoural genetic heterogeneity. The data were compared to findings on 32 post-Chernobyl PTCs that occurred after a longer period of time (9-12 years) after the accident. In none of the cases from either group were 100% of cells positive for RET rearrangement. In addition, the pattern of RET-positive cells was different in the two groups (short vs longer latency). A significant clustering of aberrant cells could be detected in the long-latency subgroup, whereas the aberrant cells were more homogeneously distributed among the short-latency tumours. The findings suggest that oligoclonal tumour development occurs in post-Chernobyl PTCs. This pattern of different clones within the tumour appears to become more discrete in cases with longer latencies, suggesting either outgrowth of individual clones or development of later subclones with time. AU - Unger, K. AU - Zurnadzhy, L.* AU - Walch, A.K. AU - Mall, M. AU - Bogdanova, T.* AU - Braselmann, H. AU - Hieber, L. AU - Tronko, N.* AU - Hutzler, P. AU - Jeremiah, S.* AU - Thomas, G.* C1 - 4508 C2 - 23966 SP - 1472-1477 TI - RET rearrangements in post-Chernobyl papillary thyroid carcinomas with a short latency analysed by interphase FISH. JO - Br. J. Cancer VL - 94 IS - 10 PY - 2006 SN - 0007-0920 ER - TY - JOUR AB - Recent analyses based on UK data indicate that people who stop smoking, even well into middle age, avoid most of their subsequent risk of lung cancer. We investigated whether similar absolute risks of lung cancer in men are found in other European countries with different smoking patterns and at different stages of their lung cancer epidemic. Using data for men from a multicentre case–control study of lung cancer in the UK, Germany, Italy and Sweden, and including 6523 lung cancer cases and 9468 controls, we combined odds ratio estimates with estimates of national lung cancer incidence rates to calculate the cumulative risk of lung cancer among men by age 75. Lung cancer cumulative risks by age 75 among continuing smokers were similar for the UK, Germany and Italy at 15.7, 14.3 and 13.8% respectively, whereas the cumulative risk among Swedish male smokers was 6.6%. The proportion of the risk of lung cancer avoided by quitting smoking before the age of 40 was comparable between the four countries, at 80% in Italy and 91% in the UK, Germany and Sweden. Similarly, the proportion of the excess risk avoided by quitting before the age of 50 ranged from 57% in Italy to 69% in Germany. Our results support the important conclusion that for long-term smokers, giving up smoking in middle age avoids most of the subsequent risk of lung cancer, and that lung cancer mortality in European men over the next three decades will be determined by the extent to which current smokers can successfully quit smoking. AU - Crispo, A.* AU - Brennan, P.* AU - Köckel, K.-H.* AU - Schaffrath Rosario, A. AU - Wichmann, H.-E. AU - Nyberg, F.* AU - Simonato, L.* AU - Merletti, F.* AU - Forastiere, F.* AU - Boffetta, P.* AU - Darby, S.* C1 - 3181 C2 - 22177 SP - 1280-1286 TI - The cumulative risk of lung cancer among current, ex- and never-smokers in European men. JO - Br. J. Cancer VL - 91 PY - 2004 SN - 0007-0920 ER - TY - JOUR AB - Epidemiological studies of lung cancer among nonsmoking men are few, This case-control study was conducted among lifetime nonsmoking men between 1990 and 1996 in Germany to examine lung cancer risk in relation to occupation, environmental tobacco smoke, residential radon, family history of cancer and previous lung disease. A total of 58 male cases with confirmed primary lung cancer and 803 male population controls who had never smoked more than 400 cigarettes in their lifetime were personally interviewed by a standardized questionnaire. In addition, 1-year radon measurements in the living and bedroom of the subjects' last dwelling were carried out. Unconditional logistic regression was used to calculate odds ratios (OR) and 95% confidence intervals (CI). Having ever worked in a job with known lung carcinogens was associated with a two-fold significantly increased lung cancer risk (OR = 2.2; CI = 1.0-5.0), adjusted for age and region. The linear trend test for lung-cancer risk associated with radon exposure was close to statistical significance, demonstrating an excess relative risk for an increase in exposure of 100 Bq m(-3) of 0.43 (P= 0.052). Nonsignificantly elevated effects of exposure to environmental tobacco smoke in public transportation and in social settings were observed, No associations with a family history of cancer or previous lung diseases were found. Our results indicate that occupational carcinogens and indoor radon may play a role in some lung cancers in nonsmoking men. AU - Kreuzer, M.* AU - Gerken, M. AU - Kreienbrock, L. AU - Wellmann, J. AU - Wichmann, H.-E. C1 - 10324 C2 - 19876 SP - 134-140 TI - Lung cancer in lifetime nonsmoking men : Results of a case-control study in Germany. JO - Br. J. Cancer VL - 84 IS - 1 PB - Nature Publishing Group PY - 2001 SN - 0007-0920 ER - TY - JOUR AB - Several studies in the past have shown appreciably higher lung cancer risk estimates associated with smoking exposure among men than among women, while more recent studies in the USA report just the opposite. To evaluate this topic in a European population we conducted a case-control study of lung cancer in three German and three Italian centres. Personal interviews and standardized questionnaires were used to obtain detailed life-long smoking and occupational histories from 3723 male and 900 female cases and 4075 male and 1094 female controls. Lung cancer risk comparing ever-smokers with never-smokers was higher among men (odds ratios (OR) adjusted for age and centre = 16.1, 95% confidence interval (CI) 12.8-20.3) than among women (OR = 4.2, CI 3.5-5.1). Because the smoking habits of women were different from men, we conducted more detailed analyses using comparable levels of smoking exposure. After restriction to smokers and adjustment for other smoking variables, risk estimates did not differ appreciably between genders. The analysis of duration of smoking (0-19, 20-39, 40+ years) adjusted for cigarette consumption and time since quitting smoking revealed similar risk estimates in men (OR = 1.0, 3.3 [CI 2.6-4.2], 4.1 [CI 3.1-5.6]) and women (OR = 1.0, 2.7 [CI 1.7-4.1], 3.3 [CI 1.9-5.8]). The same was true of the analysis of average or cumulative smoking consumption, and also of analyses stratified by different histological types. We conclude that for comparable exposure to tobacco smoke, the risk of lung cancer is comparable in women and men. AU - Kreuzer, M. AU - Boffetta, P.* AU - Whitley, E.* AU - Ahrens, W.* AU - Heinrich, J. AU - Jöckel, K.-H.* AU - Kreienbrock, L. AU - Mallone, S.* AU - Merletti, F.* AU - Roesch, F. C1 - 21323 C2 - 19438 SP - 227-233 TI - Gender differences in lung cancer risk by smoking: A multicentre case-control study in Germany and Italy. JO - Br. J. Cancer VL - 82 IS - 1 PY - 2000 SN - 0007-0920 ER - TY - JOUR AB - Bispecific antibodies (bsAb) are considered as promising tools for the elimination of disseminated tumour cells in a minimal residual disease situation. The bsAb-mediated recruitment of an immune effector cell in close vicinity of a tumour cell is thought to induce an antitumoural immune response. However, classical bispecific molecules activate only a single class of immune effector cell that may not yield optimal immune responses. We therefore constructed an intact bispecific antibody, BiUII (anti-CD3 × anti-EpCAM), that not only recognizes tumour cells and T lymphocytes with its two binding arms, but also binds and activates Fcγ-receptor positive accessory cells through its Fc-region. We have demonstrated recently that activated accessory cells contribute to the bsAb-induced antitumoural activity. We now analyse this stimulation in more detail and demonstrate here the BiUll-induced upregulation of activation markers like CD83 and CD95 on accessory cells and the induction of neopterin and biopterin synthesis. Experiments with pure cell subpopulations revealed binding of BiUll to CD64+ accessory cells and CD16+ NK cells, but not to CD32+ B lymphocytes. We provide further evidence for the importance of the Fc-region in that this bispecific molecule stimulates Fcγ-R-positive accessory cells to eliminate tumour cells in vitro by direct phagocytosis. AU - Zeidler, R.* AU - Mysliwietz, J. AU - Csanady, J.* AU - Walz, A.* AU - Ziegler, I. AU - Schmitt, B.* AU - Wollenberg, B.* AU - Lindhofer, H.* C1 - 21389 C2 - 19507 SP - 261-266 TI - The Fc-region of a new class of intact bispecific antibody mediates activation of accessory cells and NK cells and induces direct phagocytosis of tumor cells. JO - Br. J. Cancer VL - 83 PY - 2000 SN - 0007-0920 ER - TY - JOUR AB - The thyroid dose due to 131I releases during the Chernobyl accident was reconstructed for children and adolescents in two cities and 2122 settlements in Belarus, and in one city and 607 settlements in the Bryansk district of the Russian Federation. In this area, which covers the two high contamination spots in the two countries following the accident, data on thyroid cancer incidence during the period 1991-1995 were analysed in the light of possible increased thyroid surveillance. Two methods of risk analysis were applied: Poisson regression with results for the single settlements and Monte Carlo (MC) calculations for results in larger areas or sub-populations. Best estimates of both methods agreed well. Poisson regression estimates of 95% confidence intervals (CIs) were considerably smaller than the MC results, which allow for extra-Poisson uncertainties due to reconstructed doses and the background thyroid cancer incidence. The excess absolute risk per unit thyroid dose (EARPD) for the birth cohort 1971-1985 by the MC analysis was 2.1 (95% CI 1.0-4.5) cases per 10(4) person-year Gy. The point estimate is lower by a factor of two than that observed in a pooled study of thyroid cancer risk after external exposures. The excess relative risk per unit thyroid dose was 23 (95% CI 8.6-82) Gy(-1). No significant differences between countries or cities and rural areas were found. In the lowest dose group of the settlements with an average thyroid dose of 0.05 Gy the risk was statistically significantly elevated. Dependencies of risks on age-at-exposure and on gender are consistent with findings after external exposures. AU - Jacob, P. AU - Kenigsberg, Y.* AU - Goulko, G. AU - Buglova, E.* AU - Heidenreich, W.F. AU - Golovneva, A.* AU - Bratilova, A.A.* AU - Drozdovitch, V.* C1 - 20951 C2 - 19006 SP - 1461-1469 TI - Childhood exposure due to the Chernobyl accident and thyroid cancer risk in contaminated areas of Belarus and Russia. JO - Br. J. Cancer VL - 80 IS - 9 PY - 1999 SN - 0007-0920 ER - TY - JOUR AB - Cisplatin resistant mouse fibrosarcoma cells were isolated after fractionated irradiation in the absence of any drug treatment. Several sublines have been established; clone SSK-rad1 was used for further studies. These cells exhibit unchanged radiosensitivity and are compared to cisplatin resistant sublines, SSK-cis2, previously induced by low dose cisplatin exposure. Both resistant sublines are characterised by reduced CdCl2 sensitivity, indicating enhanced metallothionein content; loss of cisplatin resistance occurs after 10 to 25 generations and correlates with rising CdCl2 toxicity. Increase of MT is demonstrated directly by 109Cd binding to the MT containing region after FPLC. Both sublines differ in GSH level, which is increased only in SSK-rad1 cells, and in cellular platinum content, which is reduced in SSK-cis2 cells compared to the parental SSK cell line. These factors may contribute to cisplatin resistance but are not the main cause responsible for the transient nature of the drug resistance observed. Our results indicate that transient cisplatin resistance in SSK cells can be induced not only by the drug itself but also by gamma-irradiation and is based on the same mechanism of increased cellular MT content. AU - Eichholtz-Wirth, H. AU - Reidel, G. AU - Hietel, B. C1 - 20653 C2 - 13868 SP - 1001-1006 TI - Radiation-induced transient cisplatin resistance in murine fibrosarcoma cells associated with elevated metallothionein content. JO - Br. J. Cancer VL - 67 IS - 5 PY - 1993 SN - 0007-0920 ER - TY - JOUR AB - Cisplatin resistant mouse fibrosarcoma cells were isolated after fractionated irradiation in the absence of any drug treatment. Several sublines have been established; clone SSK-rad1 was used for further studies. These cells exhibit unchanged radiosensitivity and are compared to cisplatin resistant sublines, SSK-cis2, previously induced by low dose cisplatin exposure. Both resistant sublines are characterised by reduced CdCl2 sensitivity, indicating enhanced metallothionein content; loss of cisplatin resistance occurs after 10 to 25 generations and correlates with rising CdCl2 toxicity. Increase of MT is demonstrated directly by 109Cd binding to the MT containing region after FPLC. Both sublines differ in GSH level, which is increased only in SSK-rad, cells, and in cellular platinum content, which is reduced in SSK-cis2 cells compared to the parental SSK cell line. These factors may contribute to cisplatin resistance but are not the main cause responsible for the transient nature of the drug resistance observed. Our results indicate that transient cisplatin resistance in SSK cells can be induced not only by the drug itself but also by γ-irradiation and is based on the same mechanism of increased cellular MT content. AU - Eichholtz-Wirth, H. AU - Reidel, G.* AU - Hietel, B. C1 - 40355 C2 - 40043 SP - 1001-1006 TI - Radiation-induced transient cisplatin resistance in murine fibrosarcoma cells associated with elevated metallothionein content. JO - Br. J. Cancer VL - 67 IS - 5 PY - 1993 SN - 0007-0920 ER - TY - JOUR AB - The study was originally set up to measure accurate cell kinetic parameters in two murine squamous cell carcinomas (scc) for comparison with radiobiological data on proliferation during radiotherapy. The tumours, AT84 and AT478, were both moderately well differentiated aneuploid scc. In the course of the study, several comparisons of techniques were made in two different centres. This paper reports on the results of those comparisons involving two different detection methods (flow cytometry and immunohistochemistry), single vs double labelling, and in vivo and in vitro labelling, the latter using tissue slices incubated under high pressure oxygen. Pulse labelling studies with bromodeoxyuridine (BrdUrd) showed that the labelling indices (LI) were not significantly different after in vitro or in vivo labelling. In addition, the flow cytometry (FCM) and immunohistochemistry (IHC) methods also gave labelling indices which were not significantly different. Only tumour cells were analysed in these studies by selecting cells on the basis of aneuploidy (FCM) or morphology (IHC). The DNA synthesis time of the tumour cells were analysed by both techniques. For FCM, the Relative Movement method was used. For IHC, a double labelling method was used employing BrdUrd and triated thymidine (3H-TdR) administered several hours apart, detected simultaneously using immunoperoxidase and autoradiography, respectively. When both labels were administered in vivo, there was good agreement for T(s) between the FCM and IHC methods. Attempts were also made to measure T(s) in vitro using both techniques. With double labelling, it was found that cells did not take up the second label, implying a failure of cycle progression. This was confirmed by FCM results, showing no movement of labelled cells through the S-phase, despite an initially high uptake. This could not be influenced by lowering the DNA precursor concentration or by adding foetal calf serum. This indicates that DNA synthesis times are difficult or impossible to measure in vitro in fresh tumour explants. Finally, the double labelling IHC method allowed intratumoural variations of both LI and T(s) to be studied. Both parameters were found to vary markedly throughout the tumour volume, particularly for larger tumours (600 mg), giving calculated local potential doubling time values (T(pot)) ranging from 1-7 days. AU - Schultz-Hector, S. AU - Begg, A.C. AU - Hofland, I.D. AU - Kummermehr, J.C. AU - Sund, M. C1 - 33784 C2 - 40086 SP - 1097-1103 TI - Cell kinetic analysis of murine squamous cell carcinomas: A comparison of single versus double labelling using flow cytometry and immunohistochemistry. JO - Br. J. Cancer VL - 68 IS - 6 PY - 1993 SN - 0007-0920 ER - TY - JOUR AB - The mechanism of down-regulation of c-myc RNA associated with androgen-induced suppression of the transformed phenotype in the human prostate carcinoma cell line LNCaP was investigated. The synthetic androgen mibolerone (7 alpha-17 alpha-Dimethyl-19-nortestosterone) reversibly inhibits the proliferation of LNCaP cells and, from 12-72 h after hormone addition reduces the level of c-myc transcripts to a few per cent of controls. P1, P2, and P0 c-myc transcripts decline at the same rate, whereas P3 transcripts are much less hormone sensitive. Nuclear run-on analysis revealed that c-myc is down-regulated at the level of transcription initiation in LNCaP cells. The level of c-myc transcripts prevailing in untreated control cells can be restored in androgen-induced cells by excess antiandrogen, indicating the involvement of the androgen receptor in c-myc down-regulation. AU - Wolf, D.A. AU - Kohlhuber, F. AU - Schulz, P. AU - Fittler, F. AU - Eick, D. C1 - 19009 C2 - 12049 SP - 376-382 TI - Transcriptional Down-regulation of c-myc in Human Prostate Carcinoma Cells by the Synthetic Androgen Mibolerone. JO - Br. J. Cancer VL - 65 IS - 3 PY - 1992 SN - 0007-0920 ER - TY - JOUR AB - Clonogenic survival of HeLa, Chinese hamster and HaK cells after treatment with a range of cisplatin concentrations and exposure times was determined and cellular platinum concentrations were measured by PIXE. It was demonstrated that cisplatin cytotoxicity of the three cell lines varied considerably as a function of drug exposure dose. These differences are related to differential cellular drug uptake. AU - Eichholtz-Wirth, H. AU - Hietel, B. C1 - 33233 C2 - 35408 SP - 239-243 TI - The relationship between cisplatin sensitivity and drug uptake into mammalian cells in vitro. JO - Br. J. Cancer VL - 54 IS - 2 PY - 1986 SN - 0007-0920 ER - TY - JOUR AB - Evidence supports the view that double-strand breaks (dsb) in the DNA of X-irradiated mammalian cells are the lesions leading to chromosome aberrations and eventual cell death. The detailed kinetics of repair of dsb in Ehrlich ascites tumour cells over long repair intervals have therefore been studied and compared under conditions simulating procedures known to cause large changes in cell survival. These conditions are: holding cells in stationary phase for 7 h after X-irradiation, transference of cells to fresh growth medium immediately after X-irradiation, and treatment with the DNA synthesis inhibitor 9-β-D-arabinofuranosyladenine (ara A) for 30 min before, during and for 7 h after X-irradiation. These conditions have also been investigated for their effects on the frequencies of chromosome abnormalities (anaphase bridges and fragments). It is shown that conditions leading to both an inhibition of dsb repair (in the presence of ara A) as well as an acceleration of dsb repair (by fresh growth medium) lead to higher frequencies of chromosome abnormalities as compared to those for cells under stationary conditions for 7 h after irradiation. Holding dsb open for long periods with ara A may maximize the probablity of formation of aberrations; however, the data for fresh medium treatment show that it is not merely the rate at which dsb repair which determines the aberration frequency, and indicate the presence of other biochemical mechanisms in the cell which determine the frequency of conversion of dsb into chromosome aberrations. AU - Bryant, P.E. C1 - 40893 C2 - 38469 SP - 61-65 TI - Effects of ara A and fresh medium on chromosome damage and DNA double-strand break repair in X-irradiated stationary cells. JO - Br. J. Cancer VL - 49 IS - SUPPL. 6 PY - 1984 SN - 0007-0920 ER - TY - JOUR AB - Evidence is presented that in yeast cells one DNA double-strand break (dsb) may be considered as one potentially lethal lesion (PLL.) Using a temperature conditional radiosensitive diploid yeast mutant (rad 54-3) it is demonstrated that the shoulder of survival curves, for cells plated immediately, is due to repair of dsb (PLL) within a restricted time period. Split dose experiments with the mutant rad 54-3 show that the reappearance of a shoulder is observed when two conditions are met: (1) repair of dsb (PLL) during the time interval between doses and (2) repair of mainly those dsb (PLL) which are induced by the second dose on the nutrient agar plate. Irradiation of wild type yeast cells at high dose rate followed by liquid holding treatment for 72 h (delayed plating, DP) or at low dose rate show that the bending of DP-survival curves is due to the accumulation of dsb (PLL) leading to lethal lesions probably by misrepair of dsb. AU - Frankenberg, D. AU - Frankenberg-Schwager, M. AU - Harbich, R. C1 - 41567 C2 - 38656 SP - 233-238 TI - Interpretation of the shape of survival curves in terms of induction and repair/misrepair of DNA double-strand breaks. JO - Br. J. Cancer VL - 49 PY - 1984 SN - 0007-0920 ER - TY - JOUR AB - The role of repair of DNA double-strand breaks (dsb) in the determination of the RBE-value of alpha particles was studied using the temperature conditional radiosensitive diploid yeast mutant rad 54-3. This mutant is proficient in the repair of dsb at the permissive temperature of 23°C at which it yields a shouldered survival curve, but it is dsb repair-deficient at the restrictive temperature of 36°C at which it yields an exponential survival curve. At the permissive temperature the rad 54-3 mutant also shows liquid holding recovery of colony forming ability as a function of the liquid holding period. Thus, with this mutant it is possible to obtain survival curves involving no repair of dsb (immediate plating, 36°C), partial repair of dsb (immediate plating, 23°C) and gradually increasing levels of dsb repair by delayed plating after liquid holding periods of 24, 48 and 72 h. The RBE-values of densely ionizing 3.5 MeV alpha particles for cell killing relative to sparsely ionizing 30 MeV electrons have been determined as a function of the level of dsb repair. It is shown that the RBE-value is low and independent of dose when no repair of dsb is involved, whereas it becomes gradually larger with a gradual increase in the level of dsb repair. AU - Frankenberg-Schwager, M. AU - Frankenberg, D. AU - Harbich, R. C1 - 41744 C2 - 38408 SP - 169-173 TI - Repair of DNA double-strand breaks as a determinant of RBE of alpha particles. JO - Br. J. Cancer VL - 49 IS - SUPPL. 6 PY - 1984 SN - 0007-0920 ER - TY - JOUR AB - In a thermoconditional mutant of mutagenic DNA repair (rev2(ts) = rad5-8) of Saccharomyces cerevisiae, recovery of survival and mutation frequencies can be monitored by incubating UV-irradiated cells in growth medium at a permissive temperature (23°C) before plating and a shift to restrictive temperature (36°C). Inhibition of protein synthesis with cycloheximide during incubation at permissive conditions blocks this REV2 dependent recovery process in stationary phase rev2(ts) cells, whereas it can be reduced but not totally abolished in exponentially growing cells. These results indicate a strict dependence on post-irradiation protein synthesis in stationary phase cells and argue for a considerable constitutive level and only limited inducibility in logarithmic phase cells. The UV inducibility of the REV2 coded function in stationary phase cells could be confirmed by analysis of the dose-response pattern of the his 5-2 reversion: in stationary phase rev2(ts) cells, the quadratic component of the biphasic linear-quadratic induction kinetics found at 23°C, which is interpreted as the consequence of induction of mutagenic repair, is eliminated at 36°C. AU - Siede, W. AU - Eckardt, F. C1 - 41056 C2 - 38419 SP - 103-106 TI - Indications for an inducible component of error-prone DNA repair in yeast. JO - Br. J. Cancer VL - 49 IS - SUPPL. 6 PY - 1984 SN - 0007-0920 ER - TY - JOUR AB - Chronically hypoxic cells were 5 times more resistant to Adriamycin (ADR) than exponentially growing oxic cells. On reoxygenation, resistance decreased slowly to reach the ADR sensitivity of oxic cells after 24 h. With increasing pH, ADR efficiency increased more in oxic than in chronically hypoxic cells. With increasing cell density, ADR efficiency decreased linearly. The differences in ADR efficiency under the various conditions were accompanied by differences in intracellular ADR uptake. Chronically hypoxic cells incorporated 1.6 times less than oxic cells; the incorporation rate at pH 6.5 was half that at pH 7.4; and at a cell density of 5x105/bottle the intracellular uptake was 6 times that at 5x106/bottle. The observed differences in uptake of ADR were not, however, sufficient to explain the differences in cytotoxicity. AU - Born, R. AU - Eichholtz-Wirth, H. C1 - 42175 C2 - 38410 SP - 241-246 TI - Effect of different physiological conditions of the action of adriamycin on Chinese hamster cells in vitro. JO - Br. J. Cancer VL - 44 IS - 2 PY - 1981 SN - 0007-0920 ER -