Selection and characterization of a DNA aptamer inhibiting coagulation factor XIa

Abstract Factor XIa (FXIa) is a serine protease that catalyzes the activation of Factor IX (FIX) in the blood coagulation cascade. FXIa and its precursor FXI are emergent therapeutic targets for the development of safer anticoagulant agents. Here, we sought a novel DNA-based agent to inhibit FXIa. T...

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Autores principales:	David A. Donkor, Varsha Bhakta, Louise J. Eltringham-Smith, Alan R. Stafford, Jeffrey I. Weitz, William P. Sheffield
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/4219d136fc514ca4a4f9663c24f5819f
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spelling	oai:doaj.org-article:4219d136fc514ca4a4f9663c24f5819f2021-12-02T12:32:18ZSelection and characterization of a DNA aptamer inhibiting coagulation factor XIa10.1038/s41598-017-02055-x2045-2322https://doaj.org/article/4219d136fc514ca4a4f9663c24f5819f2017-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-02055-xhttps://doaj.org/toc/2045-2322Abstract Factor XIa (FXIa) is a serine protease that catalyzes the activation of Factor IX (FIX) in the blood coagulation cascade. FXIa and its precursor FXI are emergent therapeutic targets for the development of safer anticoagulant agents. Here, we sought a novel DNA-based agent to inhibit FXIa. Towards this goal, an 80 base, single-stranded DNA aptamer library (containing a 40 base randomized core) was screened for FXIa-binding candidates, using ten rounds of positive and negative selection. After selection, 6 of 89 different sequences inhibited FXIa-mediated chromogenic substrate S2366 cleavage. The most active anti-FXIa aptamer had a hypervariable central sequence 5′-AACCTATCGGACTATTGTTAGTGATTTTTATAGTGT-3′ and was designated Factor ELeven Inhibitory APtamer (FELIAP). FELIAP, but not a scrambled aptamer control (SCRAPT), competitively inhibited FXIa-catalyzed S2366 cleavage, FIX activation, and complex formation with antithrombin. No effect of FELIAP on FXI activation was observed. FELIAP inhibited plasma clotting and thrombin generation assays to a significantly greater extent than SCRAPT. Immobilized FELIAP bound FXIa with strong affinity and an equilibrium binding constant (KD) in the low nanomolar range determined using surface plasmon resonance. FELIAP is the first FXIa-inhibitory aptamer to be described and constitutes a lead compound to develop related aptamers for in vivo use.David A. DonkorVarsha BhaktaLouise J. Eltringham-SmithAlan R. StaffordJeffrey I. WeitzWilliam P. SheffieldNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-13 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q David A. Donkor Varsha Bhakta Louise J. Eltringham-Smith Alan R. Stafford Jeffrey I. Weitz William P. Sheffield Selection and characterization of a DNA aptamer inhibiting coagulation factor XIa
description	Abstract Factor XIa (FXIa) is a serine protease that catalyzes the activation of Factor IX (FIX) in the blood coagulation cascade. FXIa and its precursor FXI are emergent therapeutic targets for the development of safer anticoagulant agents. Here, we sought a novel DNA-based agent to inhibit FXIa. Towards this goal, an 80 base, single-stranded DNA aptamer library (containing a 40 base randomized core) was screened for FXIa-binding candidates, using ten rounds of positive and negative selection. After selection, 6 of 89 different sequences inhibited FXIa-mediated chromogenic substrate S2366 cleavage. The most active anti-FXIa aptamer had a hypervariable central sequence 5′-AACCTATCGGACTATTGTTAGTGATTTTTATAGTGT-3′ and was designated Factor ELeven Inhibitory APtamer (FELIAP). FELIAP, but not a scrambled aptamer control (SCRAPT), competitively inhibited FXIa-catalyzed S2366 cleavage, FIX activation, and complex formation with antithrombin. No effect of FELIAP on FXI activation was observed. FELIAP inhibited plasma clotting and thrombin generation assays to a significantly greater extent than SCRAPT. Immobilized FELIAP bound FXIa with strong affinity and an equilibrium binding constant (KD) in the low nanomolar range determined using surface plasmon resonance. FELIAP is the first FXIa-inhibitory aptamer to be described and constitutes a lead compound to develop related aptamers for in vivo use.
format	article
author	David A. Donkor Varsha Bhakta Louise J. Eltringham-Smith Alan R. Stafford Jeffrey I. Weitz William P. Sheffield
author_facet	David A. Donkor Varsha Bhakta Louise J. Eltringham-Smith Alan R. Stafford Jeffrey I. Weitz William P. Sheffield
author_sort	David A. Donkor
title	Selection and characterization of a DNA aptamer inhibiting coagulation factor XIa
title_short	Selection and characterization of a DNA aptamer inhibiting coagulation factor XIa
title_full	Selection and characterization of a DNA aptamer inhibiting coagulation factor XIa
title_fullStr	Selection and characterization of a DNA aptamer inhibiting coagulation factor XIa
title_full_unstemmed	Selection and characterization of a DNA aptamer inhibiting coagulation factor XIa
title_sort	selection and characterization of a dna aptamer inhibiting coagulation factor xia
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/4219d136fc514ca4a4f9663c24f5819f
work_keys_str_mv	AT davidadonkor selectionandcharacterizationofadnaaptamerinhibitingcoagulationfactorxia AT varshabhakta selectionandcharacterizationofadnaaptamerinhibitingcoagulationfactorxia AT louisejeltringhamsmith selectionandcharacterizationofadnaaptamerinhibitingcoagulationfactorxia AT alanrstafford selectionandcharacterizationofadnaaptamerinhibitingcoagulationfactorxia AT jeffreyiweitz selectionandcharacterizationofadnaaptamerinhibitingcoagulationfactorxia AT williampsheffield selectionandcharacterizationofadnaaptamerinhibitingcoagulationfactorxia
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Selection and characterization of a DNA aptamer inhibiting coagulation factor XIa

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