Structural requirements of mono- and multivalent L-selectin blocking aptamers for enhanced receptor inhibition in vitro and in vivo

Abstract L-selectin mediates extravasation of leukocytes from the blood into the surrounding tissue during inflammation and is therefore a therapeutical target in certain overwhelming immune reactions. In this study, we characterized an L-selectin specific blocking DNA aptamer with respect to nucleo...

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Bibliographic Details
Published in:Nanomedicine 2016-05, Vol.12 (4), p.901-908
Main Authors: Riese, Sebastian B., PhD, Buscher, Konrad, MD, Enders, Sven, PhD, Kuehne, Christian, PhD, Tauber, Rudolf, MD, Dernedde, Jens, PhD
Format: Article
Language:English
Subjects:
Aptamer
Aptamers, Nucleotide - antagonists & inhibitors
Aptamers, Nucleotide - chemistry
Aptamers, Nucleotide - therapeutic use
Blood Buffy Coat - drug effects
Cell Adhesion - drug effects
Extravasation
Flow chamber
Healthy Volunteers
Humans
Inflammation - drug therapy
Inflammation - pathology
Internal Medicine
L-selectin
L-Selectin - administration & dosage
L-Selectin - chemistry
Leukocytes - drug effects
Ligands
Multivalency
Oligonucleotides - chemistry
Protein Binding
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Summary:Abstract L-selectin mediates extravasation of leukocytes from the blood into the surrounding tissue during inflammation and is therefore a therapeutical target in certain overwhelming immune reactions. In this study, we characterized an L-selectin specific blocking DNA aptamer with respect to nucleotide composition and target binding. Introduction of deletions and nucleotide exchanges resulted in an optimized DNA sequence but preservation of the IC50 in the low nanomolar range. The inhibitory potential was significantly increased when the aptamer was displayed as a di- and trimer connected via appropriate linker length. Similar to monoclonal antibodies, trimer yielded picomolar IC50 values in a competitive binding assay. In comparison to the monovalent aptamer, the trivalent assembly reduced PBMC interactions to L-selectin ligands 90-fold under shear and exerted superior inhibition of PBMC rolling in vivo . In conclusion, our work demonstrates the feasibility of optimizing aptamer sequences and shows that multivalent ligand presentation enables superior adhesion receptor targeting.
ISSN:1549-9634
1549-9642
DOI:10.1016/j.nano.2015.12.379