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

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• 作者：Sebastian B. Riese ; PhD^a ; ¹ ; ^{sebastian.riese@charite.de" class="auth_mail" title="E-mail the corresponding author} ; Konrad Buscher ; MD^b ; ^c ; ¹ ; ^{kbuscher@uni-muenster.de" class="auth_mail" title="E-mail the corresponding author} ; Sven Enders ; PhD^a ; ^{sven.enders@gmx.de" class="auth_mail" title="E-mail the corresponding author} ; Christian Kuehne^a ; ^{christian.kuehne@charite.de" class="auth_mail" title="E-mail the corresponding author} ; Rudolf Tauber ; MD^a ; ^{rudolf.tauber@charite.de" class="auth_mail" title="E-mail the corresponding author} ; Jens Dernedde ; PhD^a ; ^{jens.dernedde@charite.de" class="auth_mail" title="E-mail the corresponding author}
• 关键词：DiO ; 3 ; 3&prime ; -dioctadecyloxacarbocyanine perchlorate ; Fc ; fragment crystallizable ; FITC ; fluorescein isothiocyanate ; i.a. ; intraarterial ; i.p. ; intraperitoneal ; ketamine ; (RS)-2-(2-chlorphenyl)-2-(methylamino)cyclohexanone ; PBMC ; peripheral blood mononuclear cell ; PEG ; polyethylene glycol ; PNA ; peptide nucleic acid ; PNAd ; peripheral node addressin ; PSGL-1 ; P-selectin glycoprotein ligand-1 ; sLex ; sialyl Lewis x ; sTyr ; sulfo tyrosine ; TAMRA ; 5-carboxytetramethylrhodamine ; xylazine ; N-(2 ; 6-dimeth
• 刊名：Nanomedicine: Nanotechnology, Biology and Medicine
• 出版年：2016
• 出版时间：May 2016
• 年：2016
• 卷：12
• 期：4
• 页码：901-908
• 全文大小：737 K

文摘

L-selectin mediates extravasation of leukocytes from 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.

From the Clinical Editor

During inflammation, leukocytes extravasate from blood vessels under chemotaxic signals. The presence of L-selectin on endothelium acts as a mediator for the extravasation process. In this study, the authors investigated an L-selectin specific blocking DNA aptamer in various forms, as inhibitors to leukocyte binding and extravasation. This new approach confirmed the potential use of aptamers in clinical setting.