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2. Correction: SNiPER: a novel hypermethylation biomarker panel for liquid biopsy based early breast cancer detection

Author

Mijnes, Jolein, primary, Tiedemann, Janina, additional, Eschenbruch, Julian, additional, Gasthaus, Janina, additional, Bringezu, Sarah, additional, Bauerschlag, Dirk, additional, Maass, Nicolai, additional, Arnold, Norbert, additional, Weimer, Jörg, additional, Anzeneder, Tobias, additional, Fasching, Peter A., additional, Rübner, Matthias, additional, Bruno, Benjamin, additional, Heindrichs, Uwe, additional, Freres, Jennifer, additional, Schulz, Hanna, additional, Hilgers, Ralf-Dieter, additional, Ortiz-Brüchle, Nadina, additional, Serenyi, Sonja von, additional, Knüchel, Ruth, additional, Kloten, Vera, additional, and Dahl, Edgar, additional

Published
2020
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3. Technical Evaluation of Commercial Mutation Analysis Platforms and Reference Materials for Liquid Biopsy Profiling

Author

Weber, Sabrina, primary, Spiegl, Benjamin, additional, Perakis, Samantha O., additional, Ulz, Christine M., additional, Abuja, Peter M., additional, Kashofer, Karl, additional, van der Leest, Paul, additional, Azpurua, Maria Aguirre, additional, Tamminga, Menno, additional, Brudzewsky, Dan, additional, Rothwell, Dominic G., additional, Mohan, Sumitra, additional, Sartori, Alexander, additional, Lampignano, Rita, additional, Konigshofer, Yves, additional, Sprenger-Haussels, Markus, additional, Wikman, Harriet, additional, Bergheim, Inger R., additional, Kloten, Vera, additional, Schuuring, Ed, additional, Speicher, Michael R., additional, and Heitzer, Ellen, additional

Published
2020
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4. Multicenter Evaluation of Independent High-Throughput and RT-qPCR Technologies for the Development of Analytical Workflows for Circulating miRNA Analysis

Author

Babayan, Anna, primary, Neumann, Martin H. D., additional, Herdean, Andrei, additional, Shaffer, Jonathan M., additional, Janning, Melanie, additional, Kobus, Franca, additional, Loges, Sonja, additional, Di Pasquale, Francesca, additional, Kubista, Mikael, additional, Schlumpberger, Martin, additional, Lampignano, Rita, additional, Krahn, Thomas, additional, Schlange, Thomas, additional, Sprenger-Haussels, Markus, additional, Pantel, Klaus, additional, and Kloten, Vera, additional

Published
2020
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6. Multicenter Evaluation of Circulating Cell-Free DNA Extraction and Downstream Analyses for the Development of Standardized (Pre)analytical Work Flows

Author

Lampignano, Rita, primary, Neumann, Martin H.D, additional, Weber, Sabrina, additional, Kloten, Vera, additional, Herdean, Andrei, additional, Voss, Thorsten, additional, Groelz, Daniel, additional, Babayan, Anna, additional, Tibbesma, Marco, additional, Schlumpberger, Martin, additional, Chemi, Francesca, additional, Rothwell, Dominic G, additional, Wikman, Harriet, additional, Galizzi, Jean-Pierre, additional, Riise Bergheim, Inger, additional, Russnes, Hege, additional, Mussolin, Benedetta, additional, Bonin, Serena, additional, Voigt, Christine, additional, Musa, Hanny, additional, Pinzani, Pamela, additional, Lianidou, Evi, additional, Brady, Ged, additional, Speicher, Michael R, additional, Pantel, Klaus, additional, Betsou, Fay, additional, Schuuring, Ed, additional, Kubista, Mikael, additional, Ammerlaan, Wim, additional, Sprenger-Haussels, Markus, additional, Schlange, Thomas, additional, and Heitzer, Ellen, additional

Published
2019
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7. SNiPER: a novel hypermethylation biomarker panel for liquid biopsy based early breast cancer detection

Author

Mijnes, Jolein, primary, Tiedemann, Janina, additional, Eschenbruch, Julian, additional, Gasthaus, Janina, additional, Bringezu, Sarah, additional, Bauerschlag, Dirk, additional, Maass, Nicolai, additional, Arnold, Norbert, additional, Weimer, Jörg, additional, Anzeneder, Tobias, additional, Fasching, Peter A., additional, Rübner, Matthias, additional, Bruno, Benjamin, additional, Heindrichs, Uwe, additional, Freres, Jennifer, additional, Schulz, Hanna, additional, Hilgers, Ralf-Dieter, additional, Ortiz-Brüchle, Nadina, additional, Serenyi, Sonja von, additional, Knüchel, Ruth, additional, Kloten, Vera, additional, and Dahl, Edgar, additional

Published
2019
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8. ITIH5 induces a shift in TGF-beta superfamily signaling involving Endoglin and reduces risk for breast cancer metastasis and tumor death

Author

Rose, Michael, Meurer, Steffen K., Kloten, Vera, Weiskirchen, Ralf, Denecke, Bernd, Antonopoulos, Wiebke, Deckert, Martina, Knuechel, Ruth, Dahl, Edgar, Rose, Michael, Meurer, Steffen K., Kloten, Vera, Weiskirchen, Ralf, Denecke, Bernd, Antonopoulos, Wiebke, Deckert, Martina, Knuechel, Ruth, and Dahl, Edgar

Abstract

ITIH5 has been proposed being a novel tumor suppressor in various tumor entities including breast cancer. Recently, ITIH5 was furthermore identified as metastasis suppressor gene in pancreatic carcinoma. In this study we aimed to specify the impact of ITIH5 on metastasis in breast cancer. Therefore, DNA methylation of ITIH5 promoter regions was assessed in breast cancer metastases using the TCGA portal and methylation-specific PCR (MSP). We reveal that the ITIH5 upstream promoter region is particularly responsible for ITIH5 gene inactivation predicting shorter survival of patients. Notably, methylation of this upstream ITIH5 promoter region was associated with disease progression, for example, abundantly found in distant metastases. In vitro, stably ITIH5-overexpressing MDA-MB-231 breast cancer clones were used to analyze cell invasion and to identify novel ITIH5-downstream targets. Indeed, ITIH5 re-expression suppresses invasive growth of MDA-MB-231 breast cancer cells while modulating expression of genes involved in metastasis including Endoglin (ENG), an accessory TGF-beta receptor, which was furthermore co-expressed with ITIH5 in primary breast tumors. By performing in vitro stimulation of TGF-beta signaling using TGF-beta 1 and BMP-2 we show that ITIH5 triggered a TGF-beta superfamily signaling switch contributing to downregulation of targets like Id1, known to endorse metastasis. Moreover, ITIH5 predicts longer overall survival (OS) only in those breast tumors that feature high ENG expression or inversely regulated ID1 suggesting a clinical and functional impact of an ITIH5-ENG axis for breast cancer progression. Hence, we provide evidence that ITIH5 may represent a novel modulator of TGF-beta superfamily signaling involved in suppressing breast cancer metastasis.

Published
2018

9. Additional file 1: of ITIH5 mediates epigenetic reprogramming of breast cancer cells

Author

Rose, Michael, Kloten, Vera, Noetzel, Erik, Gola, Lukas, Ehling, Josef, Heide, Timon, Meurer, Steffen, Gaiko-Shcherbak, Aljona, Sechi, Antonio, Huth, Sebastian, Weiskirchen, Ralf, Klaas, Oliver, Antonopoulos, Wiebke, Lin, Qiong, Wagner, Wolfgang, JĂźrgen Veeck, Gremse, Felix, Steitz, Julia, KnĂźchel, Ruth, and Dahl, Edgar

Subjects
skin and connective tissue diseases
Abstract

Cell plasticity of ITIH5-expressing MDA-MB-231 single-cell clones. This figure shows morphological characteristics of independent MDA-MB-231 single-cell clones using phase-contrast microscopy. (DOCX 109Â kb)

Published
2017
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10. Multicenter Evaluation of Circulating Cell-Free DNA Extraction and Downstream Analyses for the Development of Standardized (Pre)analytical Work Flows.

Author

Lampignano, Rita, Neumann, Martin H. D., Weber, Sabrina, Kloten, Vera, Herdean, Andrei, Voss, Thorsten, Groelz, Daniel, Babayan, Anna, Tibbesma, Marco, Schlumpberger, Martin, Chemi, Francesca, Rothwell, Dominic G., Wikman, Harriet, Galizzi, Jean-Pierre, Bergheim, Inger Riise, Russnes, Hege, Mussolin, Benedetta, Bonin, Serena, Voigt, Christine, and Musa, Hanny

Published
2020
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11. Epigenetic loss of putative tumor suppressorSFRP3correlates with poor prognosis of lung adenocarcinoma patients

Author

Schlensog, Martin, primary, Magnus, Lara, additional, Heide, Timon, additional, Eschenbruch, Julian, additional, Steib, Florian, additional, Tator, Maximilian, additional, Kloten, Vera, additional, Rose, Michael, additional, Noetzel, Erik, additional, Gaisa, Nadine T., additional, Knüchel, Ruth, additional, and Dahl, Edgar, additional

Published
2018
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13. Liquid biopsy in colon cancer: comparison of different circulating DNA extraction systems following absolute quantification of KRAS mutations using Intplex allele-specific PCR

Author

Kloten, Vera, primary, Rüchel, Nadine, additional, Brüchle, Nadina Ortiz, additional, Gasthaus, Janina, additional, Freudenmacher, Nils, additional, Steib, Florian, additional, Mijnes, Jolein, additional, Eschenbruch, Julian, additional, Binnebösel, Marcel, additional, Knüchel, Ruth, additional, and Dahl, Edgar, additional

Published
2017
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15. ITIH5 mediates epigenetic reprogramming of breast cancer cells

Author

Rose, Michael, primary, Kloten, Vera, additional, Noetzel, Erik, additional, Gola, Lukas, additional, Ehling, Josef, additional, Heide, Timon, additional, Meurer, Steffen K., additional, Gaiko-Shcherbak, Aljona, additional, Sechi, Antonio S., additional, Huth, Sebastian, additional, Weiskirchen, Ralf, additional, Klaas, Oliver, additional, Antonopoulos, Wiebke, additional, Lin, Qiong, additional, Wagner, Wolfgang, additional, Veeck, Jürgen, additional, Gremse, Felix, additional, Steitz, Julia, additional, Knüchel, Ruth, additional, and Dahl, Edgar, additional

Published
2017
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16. Multicenter Evaluation of Circulating Plasma MicroRNA Extraction Technologies for the Development of Clinically Feasible Reverse Transcription Quantitative PCR and Next-Generation Sequencing Analytical Work Flows.

Author

Kloten, Vera, Neumann, Martin H. D., Di Pasquale, Francesca, Sprenger-Haussels, Markus, Shaffer, Jonathan M., Schlumpberger, Martin, Herdean, Andrei, Betsou, Fay, Ammerlaan, Wim, af Hällström, Taija, Serkkola, Elina, Forsman, Tarja, Lianidou, Evi, Sjöback, Robert, Kubista, Mikael, Bender, Sebastian, Lampignano, Rita, Krahn, Thomas, and Schlange, Thomas

Published
2019
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17. Die Identifizierung und Validierung neuer potentieller Blut-basierter Früherkennungs-Biomarker sowie die funktionelle Charakterisierung von ITIH5 im humanen Mammakarzinom

Author

Kloten, Vera and Dahl, Edgar

Subjects
DNA-Methylierung, Mammakarzinom, breast cancer, Tumorsuppressor-Gen, Brustkrebs, Medizin, Tumorsuppressorgen, ddc:610, tumor suppressor gene, DNA-methylation
Abstract

Mammography has become standard of care in breast cancer screening but the limitations are well-recognized resulting in an ongoing discussion about the screening necessity. Regarding these limitations of mammography in population-based screening the validation of novel minimally-invasive screening tests that could complement to mammography is an important research topic of modern medicine. There is a growing confidence that the next generation of screening tests will be based on molecular biomarkers present in bodily fluids. Determination of promoter methylation of tumor suppressor genes in the circulating free DNA (cfDNA) of serum is a rapidly growing research field in cancer detection, indicating high specificity and sensitivity of these biomarkers. Based on the largest sample collection analyzed for serum based cfDNA methylation biomarkers so far, promoter methylation of the DKK3-ITIH5 gene combination was highly significant compared to initially six analyzed candidate genes. In an independent validation set the combined ITIH5- and DKK3-methylation achieved 40% sensitivity with a high specificity of 94% (AUC: 0,673, P

Published
2014

19. Epigenetic loss of putative tumor suppressor SFRP3 correlates with poor prognosis of lung adenocarcinoma patients.

Author

Schlensog, Martin, Magnus, Lara, Heide, Timon, Eschenbruch, Julian, Steib, Florian, Tator, Maximilian, Kloten, Vera, Rose, Michael, Noetzel, Erik, Gaisa, Nadine T., Knüchel, Ruth, and Dahl, Edgar

Abstract

Secreted frizzled related protein 3 (SFRP3) contains a cysteine-rich domain (CRD) that shares homology with Frizzled CRD and regulates WNT signaling. Independent studies showed epigenetic silencing of SFRP3 in melanoma and hepatocellular carcinoma. Moreover, a tumor suppressive function of SFRP3 was shown in androgen-independent prostate and gastric cancer cells. The current study is the first to investigate SFRP3 expression and its potential clinical impact on non-small cell lung carcinoma (NSCLC). WNT signaling components present on NSCLC subtypes were preliminary elucidated by expression data of The Cancer Genome Atlas (TCGA). We identified a distinct expression signature of relevant WNT signaling components that differ between adenocarcinoma (LUAD) and squamous cell carcinoma (LUSC). Of interest, canonical WNT signaling is predominant in LUAD samples and non-canonical WNT signaling is predominant in LUSC. In line, high SFRP3 expression resulted in beneficial clinical outcome for LUAD but not for LUSC patients. Furthermore, SFRP3 mRNA expression was significantly decreased in NSCLC tissue compared to normal lung samples. TCGA data verified the reduction of SFRP3 in LUAD and LUSC patients. Moreover, DNA hypermethylation of SFRP3 was evaluated in the TCGA methylation dataset resulting in epigenetic inactivation of SFRP3 expression in LUAD, but not in LUSC, and was validated by pyrosequencing of our NSCLC tissue cohort and in vitro demethylation experiments. Immunohistochemistry confirmed SFRP3 protein downregulation in primary NSCLC and indicated abundant expression in normal lung tissue. Two adenocarcinoma gain-of-function models were used to analyze the functional impact of SFRP3 on cell proliferation and regulation of CyclinD1 expression in vitro. Our results indicate that SFRP3 acts as a novel putative tumor suppressor gene in adenocarcinoma of the lung possibly regulating canonical WNT signaling. [ABSTRACT FROM AUTHOR]

Published
2018
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22. Epigenetic loss of putative tumor suppressor SFRP3correlates with poor prognosis of lung adenocarcinoma patients

Author

Schlensog, Martin, Magnus, Lara, Heide, Timon, Eschenbruch, Julian, Steib, Florian, Tator, Maximilian, Kloten, Vera, Rose, Michael, Noetzel, Erik, Gaisa, Nadine T., Knüchel, Ruth, and Dahl, Edgar

Abstract

ABSTRACTSecreted frizzled related protein 3 (SFRP3) contains a cysteine-rich domain (CRD) that shares homology with Frizzled CRD and regulates WNT signaling. Independent studies showed epigenetic silencing of SFRP3in melanoma and hepatocellular carcinoma. Moreover, a tumor suppressive function of SFRP3 was shown in androgen-independent prostate and gastric cancer cells. The current study is the first to investigate SFRP3expression and its potential clinical impact on non-small cell lung carcinoma (NSCLC). WNT signaling components present on NSCLC subtypes were preliminary elucidated by expression data of The Cancer Genome Atlas (TCGA). We identified a distinct expression signature of relevant WNT signaling components that differ between adenocarcinoma (LUAD) and squamous cell carcinoma (LUSC). Of interest, canonical WNT signaling is predominant in LUAD samples and non-canonical WNT signaling is predominant in LUSC. In line, high SFRP3 expression resulted in beneficial clinical outcome for LUAD but not for LUSC patients. Furthermore, SFRP3mRNA expression was significantly decreased in NSCLC tissue compared to normal lung samples. TCGA data verified the reduction of SFRP3in LUAD and LUSC patients. Moreover, DNA hypermethylation of SFRP3was evaluated in the TCGA methylation dataset resulting in epigenetic inactivation of SFRP3expression in LUAD, but not in LUSC, and was validated by pyrosequencing of our NSCLC tissue cohort and in vitrodemethylation experiments. Immunohistochemistry confirmed SFRP3 protein downregulation in primary NSCLC and indicated abundant expression in normal lung tissue. Two adenocarcinoma gain-of-function models were used to analyze the functional impact of SFRP3 on cell proliferation and regulation of CyclinD1expression in vitro. Our results indicate that SFRP3acts as a novel putative tumor suppressor gene in adenocarcinoma of the lung possibly regulating canonical WNT signaling.

Published
2018
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26. Epigenetic inactivation of the novel candidate tumor suppressor gene ITIH5in colon cancer predicts unfavorable overall survival in the CpG island methylator phenotype

Author

Kloten, Vera, Rose, Michael, Kaspar, Sophie, von Stillfried, Saskia, Knüchel, Ruth, and Dahl, Edgar

Abstract

Inter-α-trypsin inhibitor heavy chain 5 (ITIH5) is supposed to be involved in extracellular matrix stability and thus may play a key role in the inhibition of tumor progression. The current study is the first to analyze in depth ITIH5 expression as well as its potential clinical and functional impact in colon cancer. Based on 30 tumor and 30 adjacent normal tissues we examined ITIH5mRNA expression and promoter methylation, whose significance was further validated by independent data sets from The Cancer Genome Atlas (TCGA) platform. In addition, ITIH5 protein expression was evaluated using immunohistochemistry. ITIH5mRNA expression loss was significantly associated (P< 0.001) with hypermethylation of the ITIH5promoter in primary colon tumors. In addition, treatment of tumor cell lines with demethylating (DAC) and histone acetylating (TSA) agents induced ITIH5expression. In line, independent TCGA data revealed a significant expression loss of ITIH5,particularly in the MSI-high and CIMP-positive phenotype concordant with an increased ITIH5hypermethylation in CIMP-positive colon tumors (P< 0.001). In proximal, i.e., right-sided tumors, abundant ITIH5expression was associated with longer overall survival (OS, P= 0.049) and the CIMP-positive (P= 0.032) subgroup. Functionally, ITIH5 re-expression mediated a reduced proliferation in HCT116 and CaCo2 cells. In conclusion, our results indicate that ITIH5is a novel putative tumor suppressor gene in colon cancer with a potential impact in the CIMP-related pathway. ITIH5may serve as a novel epigenetic-based diagnostic biomarker with further clinical impact for risk stratification of CIMP-positive colon cancer patients.

Published
2014
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27. Circulating Tumor Cell PD-L1 Expression as Biomarker for Therapeutic Efficacy of Immune Checkpoint Inhibition in NSCLC.

Author

Kloten, Vera, Lampignano, Rita, Krahn, Thomas, and Schlange, Thomas

Subjects
*PROGRAMMED cell death 1 receptors, *NON-small-cell lung carcinoma, *LUNG cancer
Abstract

Over the last decade, the immune checkpoint blockade targeting the programmed death protein 1 (PD-1)/programmed death ligand 1 (PD-L1) axis has improved progression-free and overall survival of advanced non-small cell lung cancer (NSCLC) patients. PD-L1 tumor expression, along with tumor mutational burden, is currently being explored as a predictive biomarker for responses to immune checkpoint inhibitors (ICIs). However, lung cancer patients may have insufficient tumor tissue samples and the high bleeding risk often prevents additional biopsies and, as a consequence, immunohistological evaluation of PD-L1 expression. In addition, PD-L1 shows a dynamic expression profile and can be influenced by intratumoral heterogeneity as well as the immune cell infiltrate in the tumor and its microenvironment, influencing the response rate to PD-1/PD-L1 axis ICIs. Therefore, to identify subgroups of patients with advanced NSCLC that will most likely benefit from ICI therapies, molecular characterization of PD-L1 expression in circulating tumor cells (CTCs) might be supportive. In this review, we highlight the use of CTCs as a complementary diagnostic tool for PD-L1 expression analysis in advanced NSCLC patients. In addition, we examine technical issues of PD-L1 measurement in tissue as well as in CTCs. [ABSTRACT FROM AUTHOR]

Published
2019
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28. ITIH5 mediates epigenetic reprogramming of breast cancer cells

Author

Rose, Michael, Kloten, Vera, Weiskirchen, Ralf, Klaas, Oliver, Antonopoulos, Wiebke, Lin, Qiong, Wagner, Wolfgang, Veeck, Jürgen, Gremse, Felix, Steitz, Julia, Knüchel-Clarke, Ruth, Dahl, Edgar, Noetzel, Erik Alexander, Gola, Lukas, Ehling, Josef Ludger Anton, Heide, Timon, Meurer, Steffen K., Gaiko-Shcherbak, Aljona, Sechi, Antonio S., and Huth, Sebastian

Subjects
Cancer Research, Lung Neoplasms, Proteinase Inhibitory Proteins, Secretory, Breast Neoplasms, Epigenesis, Genetic, Mice, Breast cancer, Cell Line, Tumor, Animals, Humans, ITIH5, ddc:610, DAPK1, Cancer stem cells, Research, Extracellular matrix, DNA Methylation, Prognosis, Survival Analysis, Gene Expression Regulation, Neoplastic, Death-Associated Protein Kinases, Epigenetic reprogramming, Oncology, Molecular Medicine, Female, Neoplasm Transplantation
Abstract

Molecular cancer 16(1), 44 (2017). doi:10.1186/s12943-017-0610-2, Published by Biomed Central, London

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