Selection and Characterization of an α6β4 Integrin blocking DNA Aptamer
The heterodimeric laminin receptor α6β4 integrin plays a central role in the promotion of tumor cell growth, invasion, and organotropic metastasis. As an overproduction of the integrin is often linked to a poor prognosis, the inhibition of integrin α6β4 binding to laminin is of high therapeutical in...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2016-01-01
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| Series: | Molecular Therapy: Nucleic Acids |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2162253117300239 |
| _version_ | 1828475412116668416 |
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| author | Katharina Berg Tobias Lange Florian Mittelberger Udo Schumacher Ulrich Hahn |
| author_facet | Katharina Berg Tobias Lange Florian Mittelberger Udo Schumacher Ulrich Hahn |
| author_sort | Katharina Berg |
| collection | DOAJ |
| description | The heterodimeric laminin receptor α6β4 integrin plays a central role in the promotion of tumor cell growth, invasion, and organotropic metastasis. As an overproduction of the integrin is often linked to a poor prognosis, the inhibition of integrin α6β4 binding to laminin is of high therapeutical interest. Here, we report on the combination of a cell-systematic evolution of ligands by exponential enrichment and a bead-based selection resulting in the first aptamer inhibiting the interaction between α6β4 integrin and laminin-332. This Integrin α6β4-specific DNA Aptamer (IDA) inhibits the adhesion of prostate cancer cells (PC-3) to laminin-332 with an IC50 value of 149 nmol/l. The Kd value concerning the aptamer's interaction with PC-3 cells amounts to 137 nmol/l. Further characterization showed specificity to α6 integrins and a half-life in murine blood plasma of 6 hours. Two truncated versions of the aptamer retained their binding capacity, but lost their ability to inhibit the interaction between laminin-332 and PC-3 cells. Confocal laser scanning microscope studies revealed that the aptamer was internalized into PC-3-cells. Therefore, in addition to the adhesion-blocking function of this aptamer, IDA could also be applied for the delivery of siRNA, microRNA or toxins to cancer cells presenting the integrin α6β4. |
| first_indexed | 2024-12-11T06:12:23Z |
| format | Article |
| id | doaj.art-728e78fc9b454ddd85a786d724a7b1b7 |
| institution | Directory Open Access Journal |
| issn | 2162-2531 |
| language | English |
| last_indexed | 2024-12-11T06:12:23Z |
| publishDate | 2016-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Nucleic Acids |
| spelling | doaj.art-728e78fc9b454ddd85a786d724a7b1b72022-12-22T01:18:04ZengElsevierMolecular Therapy: Nucleic Acids2162-25312016-01-015C10.1038/mtna.2016.10Selection and Characterization of an α6β4 Integrin blocking DNA AptamerKatharina Berg0Tobias Lange1Florian Mittelberger2Udo Schumacher3Ulrich Hahn4MIN-Faculty, Chemistry Department, Institute for Biochemistry and Molecular Biology, University of Hamburg, Hamburg, GermanyUniversity Medical Center Hamburg-Eppendorf, University Cancer Center, Institute of Anatomy and Experimental Morphology, Hamburg, GermanyMIN-Faculty, Chemistry Department, Institute for Biochemistry and Molecular Biology, University of Hamburg, Hamburg, GermanyUniversity Medical Center Hamburg-Eppendorf, University Cancer Center, Institute of Anatomy and Experimental Morphology, Hamburg, GermanyMIN-Faculty, Chemistry Department, Institute for Biochemistry and Molecular Biology, University of Hamburg, Hamburg, GermanyThe heterodimeric laminin receptor α6β4 integrin plays a central role in the promotion of tumor cell growth, invasion, and organotropic metastasis. As an overproduction of the integrin is often linked to a poor prognosis, the inhibition of integrin α6β4 binding to laminin is of high therapeutical interest. Here, we report on the combination of a cell-systematic evolution of ligands by exponential enrichment and a bead-based selection resulting in the first aptamer inhibiting the interaction between α6β4 integrin and laminin-332. This Integrin α6β4-specific DNA Aptamer (IDA) inhibits the adhesion of prostate cancer cells (PC-3) to laminin-332 with an IC50 value of 149 nmol/l. The Kd value concerning the aptamer's interaction with PC-3 cells amounts to 137 nmol/l. Further characterization showed specificity to α6 integrins and a half-life in murine blood plasma of 6 hours. Two truncated versions of the aptamer retained their binding capacity, but lost their ability to inhibit the interaction between laminin-332 and PC-3 cells. Confocal laser scanning microscope studies revealed that the aptamer was internalized into PC-3-cells. Therefore, in addition to the adhesion-blocking function of this aptamer, IDA could also be applied for the delivery of siRNA, microRNA or toxins to cancer cells presenting the integrin α6β4.http://www.sciencedirect.com/science/article/pii/S2162253117300239aptamercell SELEXintegrinlaminin |
| spellingShingle | Katharina Berg Tobias Lange Florian Mittelberger Udo Schumacher Ulrich Hahn Selection and Characterization of an α6β4 Integrin blocking DNA Aptamer Molecular Therapy: Nucleic Acids aptamer cell SELEX integrin laminin |
| title | Selection and Characterization of an α6β4 Integrin blocking DNA Aptamer |
| title_full | Selection and Characterization of an α6β4 Integrin blocking DNA Aptamer |
| title_fullStr | Selection and Characterization of an α6β4 Integrin blocking DNA Aptamer |
| title_full_unstemmed | Selection and Characterization of an α6β4 Integrin blocking DNA Aptamer |
| title_short | Selection and Characterization of an α6β4 Integrin blocking DNA Aptamer |
| title_sort | selection and characterization of an α6β4 integrin blocking dna aptamer |
| topic | aptamer cell SELEX integrin laminin |
| url | http://www.sciencedirect.com/science/article/pii/S2162253117300239 |
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