In Vitro Selection of Cell-Internalizing DNA Aptamers in a Model System of Inflammatory Kidney Disease
Chronic kidney disease (CKD) is a progressive pathological condition marked by a gradual loss of kidney function. Treatment of CKD is most effective when diagnosed at an early stage and patients are still asymptomatic. However, current diagnostic biomarkers (e.g., serum creatinine and urine albumin)...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2017-09-01
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| Series: | Molecular Therapy: Nucleic Acids |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2162253117302044 |
| _version_ | 1818947813309939712 |
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| author | Glory Ranches Melanie Lukasser Herbert Schramek Andreas Ploner Taras Stasyk Gert Mayer Günter Mayer Alexander Hüttenhofer |
| author_facet | Glory Ranches Melanie Lukasser Herbert Schramek Andreas Ploner Taras Stasyk Gert Mayer Günter Mayer Alexander Hüttenhofer |
| author_sort | Glory Ranches |
| collection | DOAJ |
| description | Chronic kidney disease (CKD) is a progressive pathological condition marked by a gradual loss of kidney function. Treatment of CKD is most effective when diagnosed at an early stage and patients are still asymptomatic. However, current diagnostic biomarkers (e.g., serum creatinine and urine albumin) are insufficient for prediction of the pathogenesis of the disease. To address this need, we applied a cell-SELEX (systematic evolution of ligands by exponential enrichment) approach and identified a series of DNA aptamers, which exhibit high affinity and selectivity for cytokine-stimulated cells, resembling some aspects of a CKD phenotype. The cell-SELEX approach was driven toward the enrichment of aptamers that internalize via the endosomal pathway by isolating the endosomal fractions in each selection cycle. Indeed, we demonstrated co-localization of selected aptamers with lysosomal-associated membrane protein 1 (LAMP-1), a late endosomal and lysosomal marker protein, by fluorescence in situ hybridization. These findings are consistent with binding and subsequent internalization of the aptamers into cytokine-stimulated cells. Thus, our study sets the stage for applying selected DNA aptamers as theragnostic reagents for the development of targeted therapies to combat CKD. Keywords: cell-SELEX, DNA aptamers, chronic kidney disease, inflammatory kidney disease, cytokines |
| first_indexed | 2024-12-20T08:36:52Z |
| format | Article |
| id | doaj.art-e6cf803a91814233b0120d2e53b9cf7e |
| institution | Directory Open Access Journal |
| issn | 2162-2531 |
| language | English |
| last_indexed | 2024-12-20T08:36:52Z |
| publishDate | 2017-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Nucleic Acids |
| spelling | doaj.art-e6cf803a91814233b0120d2e53b9cf7e2022-12-21T19:46:32ZengElsevierMolecular Therapy: Nucleic Acids2162-25312017-09-018198210In Vitro Selection of Cell-Internalizing DNA Aptamers in a Model System of Inflammatory Kidney DiseaseGlory Ranches0Melanie Lukasser1Herbert Schramek2Andreas Ploner3Taras Stasyk4Gert Mayer5Günter Mayer6Alexander Hüttenhofer7Division of Genomics and RNomics, Biocenter, Medical University Innsbruck, Innsbruck 6020, Austria; Division of Medical Biochemistry, Biocenter, Medical University Innsbruck, Innsbruck 6020, AustriaDivision of Genomics and RNomics, Biocenter, Medical University Innsbruck, Innsbruck 6020, AustriaDivision of Nephrology and Hypertension, Department of Internal Medicine IV, Medical University Innsbruck, Innsbruck 6020, AustriaDivision of Genomics and RNomics, Biocenter, Medical University Innsbruck, Innsbruck 6020, Austria; Sandoz GmbH, Biochemiestrasse 10, Kundl 6250, AustriaDivision of Cell Biology, Biocenter, Medical University Innsbruck, Innsbruck 6020, AustriaDivision of Nephrology and Hypertension, Department of Internal Medicine IV, Medical University Innsbruck, Innsbruck 6020, Austria; Corresponding author: Gert Mayer, Division of Nephrology and Hypertension, Department of Internal Medicine IV, Medical University Innsbruck, Innsbruck 6020, Austria.Life and Medical Sciences Institute, Chemical Biology and Chemical Genetics, University of Bonn, Bonn 53115, Germany; Centre of Aptamer Research and Development, University of Bonn, Bonn 53115, GermanyDivision of Genomics and RNomics, Biocenter, Medical University Innsbruck, Innsbruck 6020, Austria; Corresponding author: Alexander Hüttenhofer, Division of Genomics and RNomics, Biocenter, Medical University Innsbruck, Innsbruck 6020, Austria.Chronic kidney disease (CKD) is a progressive pathological condition marked by a gradual loss of kidney function. Treatment of CKD is most effective when diagnosed at an early stage and patients are still asymptomatic. However, current diagnostic biomarkers (e.g., serum creatinine and urine albumin) are insufficient for prediction of the pathogenesis of the disease. To address this need, we applied a cell-SELEX (systematic evolution of ligands by exponential enrichment) approach and identified a series of DNA aptamers, which exhibit high affinity and selectivity for cytokine-stimulated cells, resembling some aspects of a CKD phenotype. The cell-SELEX approach was driven toward the enrichment of aptamers that internalize via the endosomal pathway by isolating the endosomal fractions in each selection cycle. Indeed, we demonstrated co-localization of selected aptamers with lysosomal-associated membrane protein 1 (LAMP-1), a late endosomal and lysosomal marker protein, by fluorescence in situ hybridization. These findings are consistent with binding and subsequent internalization of the aptamers into cytokine-stimulated cells. Thus, our study sets the stage for applying selected DNA aptamers as theragnostic reagents for the development of targeted therapies to combat CKD. Keywords: cell-SELEX, DNA aptamers, chronic kidney disease, inflammatory kidney disease, cytokineshttp://www.sciencedirect.com/science/article/pii/S2162253117302044 |
| spellingShingle | Glory Ranches Melanie Lukasser Herbert Schramek Andreas Ploner Taras Stasyk Gert Mayer Günter Mayer Alexander Hüttenhofer In Vitro Selection of Cell-Internalizing DNA Aptamers in a Model System of Inflammatory Kidney Disease Molecular Therapy: Nucleic Acids |
| title | In Vitro Selection of Cell-Internalizing DNA Aptamers in a Model System of Inflammatory Kidney Disease |
| title_full | In Vitro Selection of Cell-Internalizing DNA Aptamers in a Model System of Inflammatory Kidney Disease |
| title_fullStr | In Vitro Selection of Cell-Internalizing DNA Aptamers in a Model System of Inflammatory Kidney Disease |
| title_full_unstemmed | In Vitro Selection of Cell-Internalizing DNA Aptamers in a Model System of Inflammatory Kidney Disease |
| title_short | In Vitro Selection of Cell-Internalizing DNA Aptamers in a Model System of Inflammatory Kidney Disease |
| title_sort | in vitro selection of cell internalizing dna aptamers in a model system of inflammatory kidney disease |
| url | http://www.sciencedirect.com/science/article/pii/S2162253117302044 |
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