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Mol Ther Nucleic Acids. 2017 Sep 15;8:198-210. doi: 10.1016/j.omtn.2017.06.018. Epub 2017 Jun 27.

In Vitro Selection of Cell-Internalizing DNA Aptamers in a Model System of Inflammatory Kidney Disease.

Author information

1
Division of Genomics and RNomics, Biocenter, Medical University Innsbruck, Innsbruck 6020, Austria; Division of Medical Biochemistry, Biocenter, Medical University Innsbruck, Innsbruck 6020, Austria.
2
Division of Genomics and RNomics, Biocenter, Medical University Innsbruck, Innsbruck 6020, Austria.
3
Division of Nephrology and Hypertension, Department of Internal Medicine IV, Medical University Innsbruck, Innsbruck 6020, Austria.
4
Division of Genomics and RNomics, Biocenter, Medical University Innsbruck, Innsbruck 6020, Austria; Sandoz GmbH, Biochemiestrasse 10, Kundl 6250, Austria.
5
Division of Cell Biology, Biocenter, Medical University Innsbruck, Innsbruck 6020, Austria.
6
Division of Nephrology and Hypertension, Department of Internal Medicine IV, Medical University Innsbruck, Innsbruck 6020, Austria. Electronic address: gert.mayer@i-med.ac.at.
7
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, Germany.
8
Division of Genomics and RNomics, Biocenter, Medical University Innsbruck, Innsbruck 6020, Austria. Electronic address: alexander.huettenhofer@i-med.ac.at.

Abstract

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:

DNA aptamers; cell-SELEX; chronic kidney disease; cytokines; inflammatory kidney disease

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