Serum-stable RNA aptamers to urokinase-type plasminogen activator blocking receptor binding
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Published on: January 7, 2017
Daniel Miotto Dupont1, Jeppe Buur Madsen, Roland Karl Hartmann
1Department of Molecular Biology, University of Aarhus, DK-8000 Aarhus C, Denmark. dmd@mb.au.dk
Abstract:
The serine proteinase urokinase-type plasminogen activator (uPA) is widely recognized as a potential target for anticancer therapy. Its association with cell surfaces through the uPA receptor (uPAR) is central to its function and plays an important role in cancer invasion and metastasis. In the current study, we used systematic evolution of ligands by exponential enrichment (SELEX) to select serum-stable 2'-fluoro-pyrimidine-modified RNA aptamers specifically targeting human uPA and blocking the interaction to its receptor at low nanomolar concentrations. In agreement with the inhibitory function of the aptamers, binding was found to be dependent on the presence of the growth factor domain of uPA, which mediates uPAR binding. One of the most potent uPA aptamers, upanap-12, was analyzed in more detail and could be reduced significantly in size without severe loss of its inhibitory activity. Finally, we show that the uPA-scavenging effect of the aptamers can reduce uPAR-dependent endocytosis of the uPA-PAI-1 complex and cell-surface associated plasminogen activation in cell culture experiments. uPA-scavenging 2'-fluoro-pyrimidine-modified RNA aptamers represent a novel promising principle for interfering with the pathological functions of the uPA system.
Insights
Researchers developed novel RNA aptamers targeting urokinase-type plasminogen activator (uPA) to inhibit cancer cell invasion and metastasis. These aptamers effectively block uPA
Area of Science:
- Biochemistry
- Molecular Biology
- Oncology
Background:
- Urokinase-type plasminogen activator (uPA) is a serine proteinase implicated in cancer progression.
- uPA's interaction with its receptor (uPAR) is crucial for cancer invasion and metastasis.
- Targeting the uPA/uPAR system offers a potential therapeutic strategy for cancer treatment.
Purpose of the Study:
- To develop serum-stable RNA aptamers targeting human uPA.
- To investigate the aptamers' ability to inhibit uPA-mediated functions, including uPAR binding.
- To evaluate the therapeutic potential of these aptamers in cancer therapy.
Main Methods:
- Systematic Evolution of Ligands by Exponential Enrichment (SELEX) was employed to select RNA aptamers.
- 2'-fluoro-pyrimidine modification was used to enhance serum stability of the aptamers.
- Binding affinity and inhibitory activity of aptamers against uPA were assessed.
- Cell culture experiments were conducted to evaluate the aptamers' effect on uPAR-dependent endocytosis and plasminogen activation.
Main Results:
- Serum-stable RNA aptamers targeting human uPA were successfully selected.
- The aptamers demonstrated specific binding to uPA and inhibited its interaction with uPAR at low nanomolar concentrations.
- Inhibition was dependent on the growth factor domain of uPA, which mediates uPAR binding.
- A potent aptamer, upanap-12, was identified and could be truncated without significant loss of activity.
- Aptamers reduced uPAR-dependent endocytosis and cell-surface plasminogen activation in vitro.
Conclusions:
- 2'-fluoro-pyrimidine-modified RNA aptamers are effective inhibitors of uPA.
- These aptamers represent a promising novel strategy for interfering with the pathological functions of the uPA system in cancer.
- The uPA-scavenging aptamers show potential for anticancer therapy by disrupting key steps in cancer invasion and metastasis.
