Author Manuscript Published OnlineFirst on March 14, 2014; DOI: 10.1158/1078-0432.CCR-13-2819 Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
Circulating tumor cells in patients with testicular germ cell tumors Paulina Nastaly, Christian G. Ruf, Pascal Becker, et al. Clin Cancer Res Published OnlineFirst March 14, 2014.
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Author Manuscript Published OnlineFirst on March 14, 2014; DOI: 10.1158/1078-0432.CCR-13-2819 Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
Circulating tumor cells in patients with testicular germ cell tumors Paulina Nastay1†, Christian Ruf2,3†, Pascal Becker2, Natalia Bednarz-Knoll1, Magorzata Stoupiec1, Refik Kavsur1, Hendrik Isbarn3, Cord Matthies2, Walter Wagner2, Dirk Höppner4, Margit Fisch3, Carsten Bokemeyer5, Sascha Ahyai3, Friedemann Honecker5, Sabine Riethdorf1±, Klaus Pantel1± Authors’ Affiliations: 1University Medical Centre Hamburg-Eppendorf, Department of Tumour Biology, Hamburg, Germany; 2Federal Armed Forces Hospital, Department of Urology, Hamburg, Germany; 3University Medical Center HamburgEppendorf, Department of Urology, Hamburg, Germany; 4Federal Armed Forces Hospital, Department of Urology, Berlin, Germany; 5University Medical Center Hamburg-Eppendorf, Department of Oncology, Haematology and Bone Marrow Transplantation with Section Pneumology, Hamburg, Germany
†These authors contributed equally to the manuscript. ± These senior authors contributed equally to the manuscript.
Running title: Circulating tumor cells in testicular germ cell tumors
Key words: testicular germ cell tumors, metastasis, circulating tumor cells, blood, biomarker Financial Support: This work was supported by the German Ministry of Defense (C. R.) and the ERC Advanced Investigator Grant DISSECT (K.P.).
Corresponding Author: Prof. Dr. Klaus Pantel, Department of Tumor Biology, University Medical Centre Hamburg-Eppendorf, Martinistr. 52, D-20246 Hamburg, Germany. Phone: +49 40 7410 53503; Fax: +49 40 7410 55379; E-mail address: [email protected]
1 Downloaded from clincancerres.aacrjournals.org on May 28, 2014. © 2014 American Association for Cancer Research.
Author Manuscript Published OnlineFirst on March 14, 2014; DOI: 10.1158/1078-0432.CCR-13-2819 Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
Disclosure of Potential Conflicts of interest: K. P. has received honoraria from Veridex and Janssen.
Author’s Contributions: Conception and design: K.P., S.R., F.H., P.N., S.A., C.R. Development and methodology: P.N., F.H., S.R. Acquisition of data: C.R., P.B., H.I., C.M., F.H., W.W., D.H., S.A. Analysis and interpretation of data: P.N., N.B.-K., SR, FH Writing, review, and/or revision of the manuscript: P.N., C.R., F.H., S.R., K.P., C.B., N.B.-K. Administrative, technical, or material support: C.R., P.B., R.K., F.H. Study supervision: K.P, .S.R., F.H., M.F.
Word count: 4547 Tables count: 4 Figures count: 2 Supplementary material: Supplementary Table S1 Supplementary Figure S1
2 Downloaded from clincancerres.aacrjournals.org on May 28, 2014. © 2014 American Association for Cancer Research.
Author Manuscript Published OnlineFirst on March 14, 2014; DOI: 10.1158/1078-0432.CCR-13-2819 Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
Translational relevance This study shows that CTCs can be detected in the peripheral blood of about 18% of patients diagnosed with germ cell tumors, both using the established CellSearch® system, as well as a custom made assay using CTC enrichment and a combination of immunocytochemical markers. The presence of CTCs is associated with histologically more aggressive nonseminomatous tumors, advanced clinical stages, increased serum concentrations of tumor markers (AFP, HCG and LDH), and chemotherapy refractory relapse. The current study indicates the diagnostic potential of CTCs as prognostic biomarker and will stimulate future investigations, ideally with patients under both surveillance and chemotherapy and well documented follow-up to assess the clinical relevance of CTC analyses in GCTs. In addition, further characterization of CTCs in GCT patients will not only help to better understand the biology of the disease, but also offers the possibility to look for therapeutic targets in the era of targeted therapy.
3 Downloaded from clincancerres.aacrjournals.org on May 28, 2014. © 2014 American Association for Cancer Research.
Author Manuscript Published OnlineFirst on March 14, 2014; DOI: 10.1158/1078-0432.CCR-13-2819 Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
Abstract Purpose: Germ cell tumors (GCTs) represent the most frequent malignancies among young men, but little is known about circulating tumor cells (CTCs) in these tumors. Considering its heterogeneity, CTCs were investigated using two independent assays targeting germ and epithelial cell-specific markers, and results were correlated with disease stage, histology, and serum tumor markers. Experimental Design: CTCs were enriched from peripheral (PB, n=143 patients) and testicular vein blood (TVB, n=19 patients) using Ficoll density gradient centrifugation. For CTC detection, a combination of germ (anti-SALL4, anti-OCT3/4) and epithelial cell-specific (anti-keratin, anti-EpCAM) antibodies was used. In parallel, 122 corresponding PB samples were analysed using the CellSearch® system. Results: In total, CTCs were detected in 25/143 (17.5%) PB samples, whereas only 11.5% of patients were CTC-positive when considering exclusively the CellSearch® assay. The presence of CTCs in PB correlated to clinical stage (P
Circulating tumor cells in patients with testicular germ cell tumors Paulina Nastaly, Christian G. Ruf, Pascal Becker, et al. Clin Cancer Res Published OnlineFirst March 14, 2014.
Updated version Supplementary Material Author Manuscript
E-mail alerts Reprints and Subscriptions Permissions
Access the most recent version of this article at: doi:10.1158/1078-0432.CCR-13-2819 Access the most recent supplemental material at: http://clincancerres.aacrjournals.org/content/suppl/2014/03/14/1078-0432.CCR-13-2819.DC1.html Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
Sign up to receive free email-alerts related to this article or journal. To order reprints of this article or to subscribe to the journal, contact the AACR Publications Department at [email protected]. To request permission to re-use all or part of this article, contact the AACR Publications Department at [email protected].
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Author Manuscript Published OnlineFirst on March 14, 2014; DOI: 10.1158/1078-0432.CCR-13-2819 Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
Circulating tumor cells in patients with testicular germ cell tumors Paulina Nastay1†, Christian Ruf2,3†, Pascal Becker2, Natalia Bednarz-Knoll1, Magorzata Stoupiec1, Refik Kavsur1, Hendrik Isbarn3, Cord Matthies2, Walter Wagner2, Dirk Höppner4, Margit Fisch3, Carsten Bokemeyer5, Sascha Ahyai3, Friedemann Honecker5, Sabine Riethdorf1±, Klaus Pantel1± Authors’ Affiliations: 1University Medical Centre Hamburg-Eppendorf, Department of Tumour Biology, Hamburg, Germany; 2Federal Armed Forces Hospital, Department of Urology, Hamburg, Germany; 3University Medical Center HamburgEppendorf, Department of Urology, Hamburg, Germany; 4Federal Armed Forces Hospital, Department of Urology, Berlin, Germany; 5University Medical Center Hamburg-Eppendorf, Department of Oncology, Haematology and Bone Marrow Transplantation with Section Pneumology, Hamburg, Germany
†These authors contributed equally to the manuscript. ± These senior authors contributed equally to the manuscript.
Running title: Circulating tumor cells in testicular germ cell tumors
Key words: testicular germ cell tumors, metastasis, circulating tumor cells, blood, biomarker Financial Support: This work was supported by the German Ministry of Defense (C. R.) and the ERC Advanced Investigator Grant DISSECT (K.P.).
Corresponding Author: Prof. Dr. Klaus Pantel, Department of Tumor Biology, University Medical Centre Hamburg-Eppendorf, Martinistr. 52, D-20246 Hamburg, Germany. Phone: +49 40 7410 53503; Fax: +49 40 7410 55379; E-mail address: [email protected]
1 Downloaded from clincancerres.aacrjournals.org on May 28, 2014. © 2014 American Association for Cancer Research.
Author Manuscript Published OnlineFirst on March 14, 2014; DOI: 10.1158/1078-0432.CCR-13-2819 Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
Disclosure of Potential Conflicts of interest: K. P. has received honoraria from Veridex and Janssen.
Author’s Contributions: Conception and design: K.P., S.R., F.H., P.N., S.A., C.R. Development and methodology: P.N., F.H., S.R. Acquisition of data: C.R., P.B., H.I., C.M., F.H., W.W., D.H., S.A. Analysis and interpretation of data: P.N., N.B.-K., SR, FH Writing, review, and/or revision of the manuscript: P.N., C.R., F.H., S.R., K.P., C.B., N.B.-K. Administrative, technical, or material support: C.R., P.B., R.K., F.H. Study supervision: K.P, .S.R., F.H., M.F.
Word count: 4547 Tables count: 4 Figures count: 2 Supplementary material: Supplementary Table S1 Supplementary Figure S1
2 Downloaded from clincancerres.aacrjournals.org on May 28, 2014. © 2014 American Association for Cancer Research.
Author Manuscript Published OnlineFirst on March 14, 2014; DOI: 10.1158/1078-0432.CCR-13-2819 Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
Translational relevance This study shows that CTCs can be detected in the peripheral blood of about 18% of patients diagnosed with germ cell tumors, both using the established CellSearch® system, as well as a custom made assay using CTC enrichment and a combination of immunocytochemical markers. The presence of CTCs is associated with histologically more aggressive nonseminomatous tumors, advanced clinical stages, increased serum concentrations of tumor markers (AFP, HCG and LDH), and chemotherapy refractory relapse. The current study indicates the diagnostic potential of CTCs as prognostic biomarker and will stimulate future investigations, ideally with patients under both surveillance and chemotherapy and well documented follow-up to assess the clinical relevance of CTC analyses in GCTs. In addition, further characterization of CTCs in GCT patients will not only help to better understand the biology of the disease, but also offers the possibility to look for therapeutic targets in the era of targeted therapy.
3 Downloaded from clincancerres.aacrjournals.org on May 28, 2014. © 2014 American Association for Cancer Research.
Author Manuscript Published OnlineFirst on March 14, 2014; DOI: 10.1158/1078-0432.CCR-13-2819 Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
Abstract Purpose: Germ cell tumors (GCTs) represent the most frequent malignancies among young men, but little is known about circulating tumor cells (CTCs) in these tumors. Considering its heterogeneity, CTCs were investigated using two independent assays targeting germ and epithelial cell-specific markers, and results were correlated with disease stage, histology, and serum tumor markers. Experimental Design: CTCs were enriched from peripheral (PB, n=143 patients) and testicular vein blood (TVB, n=19 patients) using Ficoll density gradient centrifugation. For CTC detection, a combination of germ (anti-SALL4, anti-OCT3/4) and epithelial cell-specific (anti-keratin, anti-EpCAM) antibodies was used. In parallel, 122 corresponding PB samples were analysed using the CellSearch® system. Results: In total, CTCs were detected in 25/143 (17.5%) PB samples, whereas only 11.5% of patients were CTC-positive when considering exclusively the CellSearch® assay. The presence of CTCs in PB correlated to clinical stage (P
