Inhibition of Human Urokinase-Type Plasminogen Activator (uPA) Enzyme Activity and Receptor Binding by DNA Aptamers as Potential Therapeutics through Binding to the Different Forms of uPA
Urokinase-type plasminogen activator is widely discussed as a marker for cancer prognosis and diagnosis and as a target for cancer therapies. Together with its receptor, uPA plays an important role in tumorigenesis, tumor progression and metastasis. In the present study, systematic evolution of liga...
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| Format: | Article |
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MDPI AG
2022-04-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/9/4890 |
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| author | Nico Dreymann Julia Wuensche Wiebke Sabrowski Anja Moeller Denise Czepluch Dana Vu Van Susanne Fuessel Marcus M. Menger |
| author_facet | Nico Dreymann Julia Wuensche Wiebke Sabrowski Anja Moeller Denise Czepluch Dana Vu Van Susanne Fuessel Marcus M. Menger |
| author_sort | Nico Dreymann |
| collection | DOAJ |
| description | Urokinase-type plasminogen activator is widely discussed as a marker for cancer prognosis and diagnosis and as a target for cancer therapies. Together with its receptor, uPA plays an important role in tumorigenesis, tumor progression and metastasis. In the present study, systematic evolution of ligands by exponential enrichment (SELEX) was used to select single-stranded DNA aptamers targeting different forms of human uPA. Selected aptamers allowed the distinction between HMW-uPA and LMW-uPA, and therefore, presumably, have different binding regions. Here, uPAapt-02-FR showed highly affine binding with a K<sub>D</sub> of 0.7 nM for HMW-uPA and 21 nM for LMW-uPA and was also able to bind to pro-uPA with a K<sub>D</sub> of 14 nM. Furthermore, no cross-reactivity to mouse uPA or tissue-type plasminogen activator (tPA) was measured, demonstrating high specificity. Suppression of the catalytic activity of uPA and inhibition of uPAR-binding could be demonstrated through binding with different aptamers and several of their truncated variants. Since RNA aptamers are already known to inhibit uPA-uPAR binding and other pathological functions of the uPA system, these aptamers represent a novel, promising tool not only for detection of uPA but also for interfering with the pathological functions of the uPA system by additionally inhibiting uPA activity. |
| first_indexed | 2024-03-10T04:05:41Z |
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| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T04:05:41Z |
| publishDate | 2022-04-01 |
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| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-c325874f0a6f49eabb93a5d44d91e8252023-11-23T08:23:44ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-04-01239489010.3390/ijms23094890Inhibition of Human Urokinase-Type Plasminogen Activator (uPA) Enzyme Activity and Receptor Binding by DNA Aptamers as Potential Therapeutics through Binding to the Different Forms of uPANico Dreymann0Julia Wuensche1Wiebke Sabrowski2Anja Moeller3Denise Czepluch4Dana Vu Van5Susanne Fuessel6Marcus M. Menger7Fraunhofer Institute for Cell Therapy and Immunology (IZI), Branch Bioanalytics and Bioprocesses (IZI-BB), D-14476 Potsdam, GermanyRiNA GmbH, D-12489 Berlin, GermanyFraunhofer Institute for Cell Therapy and Immunology (IZI), Branch Bioanalytics and Bioprocesses (IZI-BB), D-14476 Potsdam, GermanyFraunhofer Institute for Cell Therapy and Immunology (IZI), Branch Bioanalytics and Bioprocesses (IZI-BB), D-14476 Potsdam, GermanyFraunhofer Institute for Cell Therapy and Immunology (IZI), Branch Bioanalytics and Bioprocesses (IZI-BB), D-14476 Potsdam, GermanyDepartment of Urology, Technische Universität Dresden, D-01307 Dresden, GermanyDepartment of Urology, Technische Universität Dresden, D-01307 Dresden, GermanyFraunhofer Institute for Cell Therapy and Immunology (IZI), Branch Bioanalytics and Bioprocesses (IZI-BB), D-14476 Potsdam, GermanyUrokinase-type plasminogen activator is widely discussed as a marker for cancer prognosis and diagnosis and as a target for cancer therapies. Together with its receptor, uPA plays an important role in tumorigenesis, tumor progression and metastasis. In the present study, systematic evolution of ligands by exponential enrichment (SELEX) was used to select single-stranded DNA aptamers targeting different forms of human uPA. Selected aptamers allowed the distinction between HMW-uPA and LMW-uPA, and therefore, presumably, have different binding regions. Here, uPAapt-02-FR showed highly affine binding with a K<sub>D</sub> of 0.7 nM for HMW-uPA and 21 nM for LMW-uPA and was also able to bind to pro-uPA with a K<sub>D</sub> of 14 nM. Furthermore, no cross-reactivity to mouse uPA or tissue-type plasminogen activator (tPA) was measured, demonstrating high specificity. Suppression of the catalytic activity of uPA and inhibition of uPAR-binding could be demonstrated through binding with different aptamers and several of their truncated variants. Since RNA aptamers are already known to inhibit uPA-uPAR binding and other pathological functions of the uPA system, these aptamers represent a novel, promising tool not only for detection of uPA but also for interfering with the pathological functions of the uPA system by additionally inhibiting uPA activity.https://www.mdpi.com/1422-0067/23/9/4890biomarkercancercancer therapyDNA aptamermicroscale thermophoresis (MST)SELEX |
| spellingShingle | Nico Dreymann Julia Wuensche Wiebke Sabrowski Anja Moeller Denise Czepluch Dana Vu Van Susanne Fuessel Marcus M. Menger Inhibition of Human Urokinase-Type Plasminogen Activator (uPA) Enzyme Activity and Receptor Binding by DNA Aptamers as Potential Therapeutics through Binding to the Different Forms of uPA International Journal of Molecular Sciences biomarker cancer cancer therapy DNA aptamer microscale thermophoresis (MST) SELEX |
| title | Inhibition of Human Urokinase-Type Plasminogen Activator (uPA) Enzyme Activity and Receptor Binding by DNA Aptamers as Potential Therapeutics through Binding to the Different Forms of uPA |
| title_full | Inhibition of Human Urokinase-Type Plasminogen Activator (uPA) Enzyme Activity and Receptor Binding by DNA Aptamers as Potential Therapeutics through Binding to the Different Forms of uPA |
| title_fullStr | Inhibition of Human Urokinase-Type Plasminogen Activator (uPA) Enzyme Activity and Receptor Binding by DNA Aptamers as Potential Therapeutics through Binding to the Different Forms of uPA |
| title_full_unstemmed | Inhibition of Human Urokinase-Type Plasminogen Activator (uPA) Enzyme Activity and Receptor Binding by DNA Aptamers as Potential Therapeutics through Binding to the Different Forms of uPA |
| title_short | Inhibition of Human Urokinase-Type Plasminogen Activator (uPA) Enzyme Activity and Receptor Binding by DNA Aptamers as Potential Therapeutics through Binding to the Different Forms of uPA |
| title_sort | inhibition of human urokinase type plasminogen activator upa enzyme activity and receptor binding by dna aptamers as potential therapeutics through binding to the different forms of upa |
| topic | biomarker cancer cancer therapy DNA aptamer microscale thermophoresis (MST) SELEX |
| url | https://www.mdpi.com/1422-0067/23/9/4890 |
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