A multi-analyte liquid biopsy approach for nonseminomatous testicular germ cell tumors: combining cfDNA and N-glycan analysis in blood and seminal plasma
Abstract Background Nonseminomatous testicular germ cell tumors (NSE) present significant diagnostic challenges, especially for the early detection of serum tumor marker (STM) negative cases. Current diagnostic tools are limited, highlighting the need for innovative approaches. This study investigat...
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| Language: | English |
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BMC
2025-07-01
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| Series: | Cancer Cell International |
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| Online Access: | https://doi.org/10.1186/s12935-025-03887-8 |
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| author | Jure Krasic Dinko Soic Lucija Skara Abramovic Ivona Kolosnjaj Miroslav Tomic Alen Vrtaric Sasa Kralik Oguic Nina Gelo Ana Katusic Bojanac Davor Jezek Dinko Mitrecic Monika Ulamec Tomislav Kulis Olga Gornik Nino Sincic |
| author_facet | Jure Krasic Dinko Soic Lucija Skara Abramovic Ivona Kolosnjaj Miroslav Tomic Alen Vrtaric Sasa Kralik Oguic Nina Gelo Ana Katusic Bojanac Davor Jezek Dinko Mitrecic Monika Ulamec Tomislav Kulis Olga Gornik Nino Sincic |
| author_sort | Jure Krasic |
| collection | DOAJ |
| description | Abstract Background Nonseminomatous testicular germ cell tumors (NSE) present significant diagnostic challenges, especially for the early detection of serum tumor marker (STM) negative cases. Current diagnostic tools are limited, highlighting the need for innovative approaches. This study investigates a novel multi-analyte approach combining circulating cell-free DNA (cfDNA) and N-glycan profiling in both blood and seminal plasma to improve NSE diagnostics. Methods The study included 41 NSE patients and 114 healthy controls. Diagnostic potential of cfDNA parameters (quality and fragmentation), cfDNA methylation (RASSF1A, PRSS21, and LINE-1) and N-glycan alterations in blood plasma and seminal plasma samples was investigated using logistic regression models. Pre- and post-radical orchidectomy longitudinal samples from NSE patients were analyzed to assess surgical treatment response and disease monitoring utility. Results Blood plasma analysis of combined cfDNA and N-glycan profiling demonstrated high diagnostic precision, with an AUC of 0.96, identifying 85% of STM-negative patients and all pure-form teratomas. Post-operative blood plasma analysis showed that LINE-1 cfDNA methylation levels returned to those of healthy controls. Seminal plasma analysis revealed an increased cfDNA fragmentation index (CFI) and cfDNA methylation changes in LINE-1 and PRSS21, with an AUC of 0.83 and identifying 85% of STM-negative patients. Conclusion The proposed multi-analyte approach significantly improves early diagnostics of NSE, particularly for STM-negative cases and teratomas. LINE-1 cfDNA methylation is a promising biomarker for NSE detection and treatment monitoring. These findings could transform diagnostic strategies and patient management in testicular germ cell tumors, with potential applications in reducing overtreatment and improving outcomes . Graphical Abstract |
| format | Article |
| id | doaj-art-13e0f0b4a78f4e1681e41bdc96caf81d |
| institution | Kabale University |
| issn | 1475-2867 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | BMC |
| record_format | Article |
| series | Cancer Cell International |
| spelling | doaj-art-13e0f0b4a78f4e1681e41bdc96caf81d2025-08-20T03:46:16ZengBMCCancer Cell International1475-28672025-07-0125111510.1186/s12935-025-03887-8A multi-analyte liquid biopsy approach for nonseminomatous testicular germ cell tumors: combining cfDNA and N-glycan analysis in blood and seminal plasmaJure Krasic0Dinko Soic1Lucija Skara Abramovic2Ivona Kolosnjaj3Miroslav Tomic4Alen Vrtaric5Sasa Kralik Oguic6Nina Gelo7Ana Katusic Bojanac8Davor Jezek9Dinko Mitrecic10Monika Ulamec11Tomislav Kulis12Olga Gornik13Nino Sincic14Department of Histology and Embryology, School of Medicine, University of ZagrebFaculty of Pharmacy and Biochemistry, University of ZagrebCentre of Excellence for Reproductive and Regenerative Medicine, School of Medicine, University of ZagrebFaculty of Pharmacy and Biochemistry, University of ZagrebCentre of Excellence for Reproductive and Regenerative Medicine, School of Medicine, University of ZagrebCentre of Excellence for Reproductive and Regenerative Medicine, School of Medicine, University of ZagrebCentre of Excellence for Reproductive and Regenerative Medicine, School of Medicine, University of ZagrebCentre of Excellence for Reproductive and Regenerative Medicine, School of Medicine, University of ZagrebCentre of Excellence for Reproductive and Regenerative Medicine, School of Medicine, University of ZagrebDepartment of Histology and Embryology, School of Medicine, University of ZagrebDepartment of Histology and Embryology, School of Medicine, University of ZagrebCentre of Excellence for Reproductive and Regenerative Medicine, School of Medicine, University of ZagrebCentre of Excellence for Reproductive and Regenerative Medicine, School of Medicine, University of ZagrebFaculty of Pharmacy and Biochemistry, University of ZagrebCentre of Excellence for Reproductive and Regenerative Medicine, School of Medicine, University of ZagrebAbstract Background Nonseminomatous testicular germ cell tumors (NSE) present significant diagnostic challenges, especially for the early detection of serum tumor marker (STM) negative cases. Current diagnostic tools are limited, highlighting the need for innovative approaches. This study investigates a novel multi-analyte approach combining circulating cell-free DNA (cfDNA) and N-glycan profiling in both blood and seminal plasma to improve NSE diagnostics. Methods The study included 41 NSE patients and 114 healthy controls. Diagnostic potential of cfDNA parameters (quality and fragmentation), cfDNA methylation (RASSF1A, PRSS21, and LINE-1) and N-glycan alterations in blood plasma and seminal plasma samples was investigated using logistic regression models. Pre- and post-radical orchidectomy longitudinal samples from NSE patients were analyzed to assess surgical treatment response and disease monitoring utility. Results Blood plasma analysis of combined cfDNA and N-glycan profiling demonstrated high diagnostic precision, with an AUC of 0.96, identifying 85% of STM-negative patients and all pure-form teratomas. Post-operative blood plasma analysis showed that LINE-1 cfDNA methylation levels returned to those of healthy controls. Seminal plasma analysis revealed an increased cfDNA fragmentation index (CFI) and cfDNA methylation changes in LINE-1 and PRSS21, with an AUC of 0.83 and identifying 85% of STM-negative patients. Conclusion The proposed multi-analyte approach significantly improves early diagnostics of NSE, particularly for STM-negative cases and teratomas. LINE-1 cfDNA methylation is a promising biomarker for NSE detection and treatment monitoring. These findings could transform diagnostic strategies and patient management in testicular germ cell tumors, with potential applications in reducing overtreatment and improving outcomes . Graphical Abstracthttps://doi.org/10.1186/s12935-025-03887-8Cell-free DNAN-glycosylationEpigeneticsBiomarkersTesticular germ cell tumorsSemen quality |
| spellingShingle | Jure Krasic Dinko Soic Lucija Skara Abramovic Ivona Kolosnjaj Miroslav Tomic Alen Vrtaric Sasa Kralik Oguic Nina Gelo Ana Katusic Bojanac Davor Jezek Dinko Mitrecic Monika Ulamec Tomislav Kulis Olga Gornik Nino Sincic A multi-analyte liquid biopsy approach for nonseminomatous testicular germ cell tumors: combining cfDNA and N-glycan analysis in blood and seminal plasma Cancer Cell International Cell-free DNA N-glycosylation Epigenetics Biomarkers Testicular germ cell tumors Semen quality |
| title | A multi-analyte liquid biopsy approach for nonseminomatous testicular germ cell tumors: combining cfDNA and N-glycan analysis in blood and seminal plasma |
| title_full | A multi-analyte liquid biopsy approach for nonseminomatous testicular germ cell tumors: combining cfDNA and N-glycan analysis in blood and seminal plasma |
| title_fullStr | A multi-analyte liquid biopsy approach for nonseminomatous testicular germ cell tumors: combining cfDNA and N-glycan analysis in blood and seminal plasma |
| title_full_unstemmed | A multi-analyte liquid biopsy approach for nonseminomatous testicular germ cell tumors: combining cfDNA and N-glycan analysis in blood and seminal plasma |
| title_short | A multi-analyte liquid biopsy approach for nonseminomatous testicular germ cell tumors: combining cfDNA and N-glycan analysis in blood and seminal plasma |
| title_sort | multi analyte liquid biopsy approach for nonseminomatous testicular germ cell tumors combining cfdna and n glycan analysis in blood and seminal plasma |
| topic | Cell-free DNA N-glycosylation Epigenetics Biomarkers Testicular germ cell tumors Semen quality |
| url | https://doi.org/10.1186/s12935-025-03887-8 |
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