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A kallikrein-targeting RNA aptamer inhibits the intrinsic pathway of coagulation and reduces bradykinin release.

Publication ,  Journal Article
Steen Burrell, K-A; Layzer, J; Sullenger, BA
Published in: J Thromb Haemost
September 2017

UNLABELLED: Essentials Kallikrein amplifies contact activation and is a potential target for preventing thrombosis. We developed and characterized a kallikrein aptamer using convergent evolution and kinetic assays. Kall1-T4 prolongs intrinsic clotting time by inhibiting factor XIIa-mediated prekallikrein activation. Kall1-T4 decreases high-molecular-weight kininogen cleavage and bradykinin release. SUMMARY: Background Plasma kallikrein is a serine protease that plays an integral role in many biological processes, including coagulation, inflammation, and fibrinolysis. The main function of kallikrein in coagulation is the amplification of activated factor XII (FXIIa) production, which ultimately leads to thrombin generation and fibrin clot formation. Kallikrein is generated by FXIIa-mediated cleavage of the zymogen prekallikrein, which is usually complexed with the non-enzymatic cofactor high molecular weight kininogen (HK). HK also serves as a substrate for kallikrein to generate the proinflammatory peptide bradykinin (BK). Interestingly, prekallikrein-deficient mice are protected from thrombotic events while retaining normal hemostatic capacity. Therefore, therapeutic targeting of kallikrein may provide a safer alternative to traditional anticoagulants with anti-inflammatory benefits. Objectives To isolate and characterize an RNA aptamer that binds to and inhibits plasma kallikrein, and to elucidate its mechanism of action. Methods and Results Using convergent Systematic Evolution of Ligands by Exponential Enrichment (SELEX), we isolated an RNA aptamer that targets kallikrein. This aptamer, Kall1-T4, specifically binds to both prekallikrein and kallikrein with similar subnanomolar binding affinities, and dose-dependently prolongs fibrin clot formation in an activated partial thromboplastin time (APTT) coagulation assay. In a purified in vitro system, Kall1-T4 inhibits the reciprocal activation of prekallikrein and FXII primarily by reducing the rate of FXIIa-mediated prekallikrein activation. Additionally, Kall1-T4 significantly reduces kallikrein-mediated HK cleavage and subsequent BK release. Conclusions We have isolated a specific and potent inhibitor of prekallikrein/kallikrein activity that serves as a powerful tool for further elucidating the role of kallikrein in thrombosis and inflammation.

Duke Scholars

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Published In

J Thromb Haemost

DOI

EISSN

1538-7836

Publication Date

September 2017

Volume

15

Issue

9

Start / End Page

1807 / 1817

Location

England

Related Subject Headings

  • Thrombosis
  • Protein Binding
  • Prekallikrein
  • Partial Thromboplastin Time
  • Kininogen, High-Molecular-Weight
  • Kinetics
  • Kallikreins
  • Humans
  • Factor XIIa
  • Dose-Response Relationship, Drug
 

Citation

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Steen Burrell, K.-A., Layzer, J., & Sullenger, B. A. (2017). A kallikrein-targeting RNA aptamer inhibits the intrinsic pathway of coagulation and reduces bradykinin release. J Thromb Haemost, 15(9), 1807–1817. https://doi.org/10.1111/jth.13760
Steen Burrell, K. -. A., J. Layzer, and B. A. Sullenger. “A kallikrein-targeting RNA aptamer inhibits the intrinsic pathway of coagulation and reduces bradykinin release.J Thromb Haemost 15, no. 9 (September 2017): 1807–17. https://doi.org/10.1111/jth.13760.
Steen Burrell K-A, Layzer J, Sullenger BA. A kallikrein-targeting RNA aptamer inhibits the intrinsic pathway of coagulation and reduces bradykinin release. J Thromb Haemost. 2017 Sep;15(9):1807–17.
Steen Burrell, K. .. A., et al. “A kallikrein-targeting RNA aptamer inhibits the intrinsic pathway of coagulation and reduces bradykinin release.J Thromb Haemost, vol. 15, no. 9, Sept. 2017, pp. 1807–17. Pubmed, doi:10.1111/jth.13760.
Steen Burrell K-A, Layzer J, Sullenger BA. A kallikrein-targeting RNA aptamer inhibits the intrinsic pathway of coagulation and reduces bradykinin release. J Thromb Haemost. 2017 Sep;15(9):1807–1817.
Journal cover image

Published In

J Thromb Haemost

DOI

EISSN

1538-7836

Publication Date

September 2017

Volume

15

Issue

9

Start / End Page

1807 / 1817

Location

England

Related Subject Headings

  • Thrombosis
  • Protein Binding
  • Prekallikrein
  • Partial Thromboplastin Time
  • Kininogen, High-Molecular-Weight
  • Kinetics
  • Kallikreins
  • Humans
  • Factor XIIa
  • Dose-Response Relationship, Drug