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DOI: 10.1055/s-0037-1614126
Blocking the Initiation of Coagulation by RNA Aptamers to Factor VIIa
We thank J. Jones and S. Daubendiek for helpful discussion, and the Duke Clinical Coagulation Lab, especially T. Ortel, A. Joyner and L. Estrada for their time, technical assistance and use of equipment and reagents. C.P.R. is a Fellow of the Jane Coffin Childs Memorial Fund for Medical Research. A. Y. was supported by a Claude E. Welch fellowship sponsored through the Department of Surgery at the Massachusetts General Hospital. J.H.L is a supported by a Clinician Scientist Award from the American Heart Association.Publikationsverlauf
Received
17. Februar 2000
Accepted after revision
31. Mai 2000
Publikationsdatum:
13. Dezember 2017 (online)
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Summary
The tissue factor/factor VIIa complex is thought to be the primary initiator of most physiologic blood coagulation events. Because of its proximal role in this process, we sought to generate new inhibitors of tissue factor/factor VIIa activity by targeting factor VIIa. We employed a combinatorial RNA library and in vitro selection methods to isolate a high affinity, nuclease-resistant RNA ligand that binds specifically to coagulation factor VII/VIIa. This RNA inhibits the tissue factordependent activation of factor X by factor VIIa. Kinetic analyses of the mechanism of action of this RNA suggest that it antagonizes factor VIIa activity by preventing formation of a functional factor VII/tissue factor complex. Furthermore, this RNA significantly prolongs the prothrombin time of human plasma in a dose dependent manner, and has an in vitro half-life of ∼15 h in human plasma. Thus, this RNA ligand represents a novel class of anticoagulant agents directed against factor VIIa.
* C. P. R. and A. Y. contributed equally to this work
* Present address: Dr. A. Yeh, Department of Surgery, Massachusetts General Hospital, Boston, MA 02115, USA
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References
- 1 Osterud B. Tissue factor: a complex biological role. Thromb Haemost 1997; 78: 755-8.
- 2 Camerer E, Kolsto AB, Prydz H. Cell biology of tissue factor, the principal initiator of blood coagulation. Thromb Res 1996; 81: 1-41.
- 3 Drake TA, Morrissey JH, Edgington TS. Selective cellular expression of tissue factor in human tissues. Implications for disorders of hemostasis and thrombosis. Am J Pathol 1989; 134: 1087-97.
- 4 Fleck RA, Rao LV, Rapaport SI, Varki N. Localization of human tissue factor antigen by immunostaining with monospecific, polyclonal anti-human tissue factor antibody. Thromb Res 1990; 59: 421-37.
- 5 Flossel C, Luther T, Muller M, Albrecht S, Kasper M. Immunohistochemical detection of tissue factor (TF) on paraffin sections of routinely fixed human tissue. Histochemistry 1994; 101: 449-53.
- 6 Nemerson Y. Tissue factor and hemostasis. Blood 1988; 71: 1-8.
- 7 Edgington TS, Mackman N, Brand K, Ruf W. The structural biology of expression and function of tissue factor. Thromb Haemost 1991; 66: 67-79.
- 8 Banner DW, D’Arcy A, Chene C, Winkler FK, Guha A, Konigsberg WH, Nemerson Y, Kirchhofer D. The crystal structure of the complex of blood coagulation factor VIIa with soluble tissue factor. Nature 1996; 380: 41-6.
- 9 Lawson JH, Butenas S, Mann KG. The evaluation of complex-dependent alterations in human factor VIIa. J Biol Chem 1992; 267: 4834-43.
- 10 Furie B, Furie BC. The molecular basis of blood coagulation. Cell 1988; 53: 505-18.
- 11 ten Cate H, Bauer KA, Levi M, Edgington TS, Sublett RD, Barzegar S, Kass BL, Rosenberg RD. The activation of factor X and prothrombin by recombinant factor VIIa in vivo is mediated by tissue factor. J Clin Invest 1993; 92: 1207-12.
- 12 Hoffman M, Monroe DM, Oliver JA, Roberts HR. Factors IXa and Xa play distinct roles in tissue factor-dependent initiation of coagulation. Blood 1995; 86: 1794-801.
- 13 Kjalke M, Monroe DM, Hoffman M, Oliver JA, Ezban M, Roberts HR. Active site-inactivated factors VIIa, Xa, and IXa inhibit individual steps in a cell-based model of tissue factor-initiated coagulation. Thromb Haemost 1998; 80: 578-84.
- 14 Hoffman M, Monroe DM, Roberts HR. Activated factor VII activates factors IX and X on the surface of activated platelets; thoughts on the mechanism of action of high-dose activated factor VII. Blood Coagul Fibrinolysis 1998; 09 (Suppl. 01) S61-5.
- 15 Lawson JH, Mann KG. Cooperative activation of human factor XI by the human extrinsic pathway of blood coagulation. J Biol Chem 1991; 266: 11317-27.
- 16 Vu TK, Hung DT, Wheaton VI, Coughlin SR. Molecular cloning of a func tional thrombin receptor reveals a novel proteolytic mechanism of receptor activation. Cell 1991; 64: 1057-68.
- 17 Harker LA, Hanson SR, Wilcox JN, Kelly AB. Antithrombotic and antilesion benefits without hemorrhagic risks by inhibiting tissue factor pathway. Haemostasis 1996; 26 (Suppl. 01) 76-82.
- 18 Johnson K, Hung D. Novel anticoagulants based on inhibition of the factor VIIa/tissue factor pathway. Coron Artery Dis 1998; 09: 83-7.
- 19 Warr TA, Rao LV, Rapaport SI. Disseminated intravascular coagulation in rabbits induced by administration of endotoxin or tissue factor: effect of anti-tissue factor antibodies and measurement of plasma extrinsic pathway inhibitor activity. Blood 1990; 75: 1481-9.
- 20 Taylor Jr FB, Chang A, Ruf W, Morrissey JH, Hinshaw L, Catlett R, Blick K, Edgington TS. Lethal E. coli septic shock is prevented by blocking tissue factor with monoclonal antibody. Circ Shock 1991; 33: 127-34.
- 21 Biemond BJ, Levi M, ten Cate H, Soule HR, Morris LD, Foster DL, Bogowitz CA, van der Poll T, Buller HR, ten Cate JW. Complete inhibition of endotoxin-induced coagulation activation in chimpanzees with a monoclonal Fab fragment against factor VII/VIIa. Thromb Haemost 1995; 73: 223-30.
- 22 Lollar P, Fass DN. Inhibition of activated porcine factor IX by dansylglutamyl-glycyl-arginyl-chloromethylketone. Arch Biochem Biophys 1984; 233: 438-46.
- 23 Bajaj SP, Rapaport SI, Maki SL. A monoclonal antibody to factor IX that inhibits the factor VIII:Ca potentiation of factor X activation. J Biol Chem 1985; 260: 11574-80.
- 24 Choudhri TF, Hoh BL, Prestigiacomo CJ, Huang J, Kim LJ, Schmidt AM, Kisiel W, Connolly Jr ES, Pinksy DJ. Targeted inhibition of intrinsic coagulation limits cerebral injury in stroke without increasing intracerebral hemorrhage. J Exp Med 1999; 190: 91-9.
- 25 Feuerstein GZ, Toomey JR, Valocik R, Koster P, Patel A, Blackburn MN. An inhibitory anti-factor IX antibody effectively reduces thrombus formation in a rat model of venous thrombosis. Thromb Haemost 1999; 82: 1443-5.
- 26 Tuerk C, Gold L. Systemtic evolution of ligands by exponential enrichment: RNA ligands to bacteriophage T4 DNA polymerase. Science 1990; 249: 505-10.
- 27 Ellington AD, Szostak JW. In vitro selection of RNA molecules that bind specific ligands. Nature 1990; 346: 818-22.
- 28 Gold L, Polisky B, Uhlenbeck O, Yarus M. Diversity of oligonucleotide functions. Annu Rev Biochem 1995; 64: 763-97.
- 29 Ishizaki J, Nevins JR, Sullenger BA. Inhibition of cell proliferation by an RNA ligand that selectively blocks E2F function. Nat Med 1996; 02: 1386-9.
- 30 Hicke BJ, Watson SR, Koenig A. et al. DNA aptamers block L-selectin function in vivo. Inhibition of human lymphocyte trafficking in SCID mice. J Clin Invest 1996; 98: 2688-92.
- 31 Floege J, Ostendorf T, Janssen U, Burg M, Radeke HH, Vargeese C, Gill SC, Green LS, Janjic N. Novel approach to specific growth factor inhibition in vivo: antagonism of platelet-derived growth factor in glomerulonephritis by aptamers. Am J Pathol 1999; 154: 169-79.
- 32 Ostendorf T, Kunter U, Eitner F, Loos A, Regele H, Kerjaschki D, Henninger DD, Janjic N, Floege J. VEGF (165) mediates glomerular endothelial repair. J Clin Invest 1999; 104: 913-23.
- 33 Bock LC, Griffin LC, Latham JA, Vermaas EH, Toole JJ. Selection of single-stranded DNA molecules that bind and inhibit human thrombin. Nature 1992; 355: 564-6.
- 34 Kubik MF, Stephens AW, Schneider D, Marlar RA, Tasset D. High-affinity RNA ligand to human alpha-thrombin. Nucleic Acids Res 1994; 22: 2619-26.
- 35 Tasset DM, Kubik MF, Steiner W. Oligonucleotide inhibitors of human thrombin that bind distinct epitopes. J Mol Biol 1997; 272: 688-98.
- 36 Griffin LC, Tidmarsh GF, Bock LC, Toole JJ, Leung LL. In vivo anticoagulant properties of a novel nucleotide-based thrombin inhibitor and demonstration of regional anticoagulation in extracorporeal circuits. Blood 1993; 81: 3271-6.
- 37 DeAnda Jr A, Coutre SE, Moon MR, Vial CM, Griffin LC, Law VS, Komeda M, Leung LL, Miller DC. Pilot study of the efficacy of a thrombin inhibitor for use during cardiopulmonary bypass. Ann Thorac Surg 1994; 58: 344-50.
- 38 Fitzwater T, Polisky B. A SELEX primer. Methods Enzymol 1996; 267: 275-301.
- 39 Lee SW, Sullenger BA. Isolation of a nuclease-resistant decoy RNA that selectively blocks autoantibody binding to insulin receptors on human lymphocytes. J Exp Med 1996; 184: 315-24.
- 40 Lee SW, Sullenger BA. Isolation of a nuclease-resistant decoy RNA that can protect human acetylcholine receptors from myasthenic antibodies. Nat Biotechnol 1997; 15: 41-5.
- 41 Wong I, Lohman TM. A double-filter method for nitrocellulose-filter binding: application to protein-nucleic acid interactions. Proc Natl Acad Sci USA 1993; 90: 5428-32.
- 42 Bevliacqua PC, Cech TR. Minor-groove recognition of double-stranded RNA by the double-stranded RNA-binding domain from the RNA-activated protein kinase PKR. Biochemistry 1996; 35: 9983-94.
- 43 Lawson JH, Krishnaswamy S, Butenas S, Mann KG. Extrinsic pathway proteolytic activity. Methods Enzymol 1993; 222: 177-95.
- 44 Williams JW, Morrison JF. The kinetics of reversible tight-binding inhibition. Methods Enzymol 1979; 63: 437-67.
- 45 Hirsh J, Dalen JE, Anderson DR, Poller L, Bussey H, Ansell J, Deykin D, Brandt JT. Oral anticoagulants: mechanism of action, clinical effectiveness, and optimal therapeutic range. Chest 1998; 114: 445S-69S.
- 46 Beigelman L, McSwiggen JA, Draper KG. et al. Chemical modification of hammerhead ribozymes. Catalytic activity and nuclease resistance. J Biol Chem 1995; 270: 25702-8.
- 47 Pieken WA, Olsen DB, Benseler F, Aurup H, Eckstein F. Kinetic characterization of ribonuclease-resistant 2’-modified hammerhead ribozymes. Science 1991; 253: 314-7.
- 48 Mathews DH, Sabina J, Zuker M, Turner DH. Expanded sequence dependence of thermodynamic parameters improves prediction of RNA secondary structure. J Mol Biol 1999; 288: 911-40.
- 49 Kelley RF, Refino CJ, O’Connell MP, Modi N, Sehl P, Lowe D, Pater C, Bunting S. A soluble tissue factor mutant is a selective anticoagulant and antithrombotic agent. Blood 1997; 89: 3219-27.
- 50 Kalafatis M, Swords NA, Rand MD, Mann KG. Membrane-dependent reactions in blood coagulation: role of the vitamin K-dependent enzyme complexes. Biochim Biophys Acta 1994; 1227: 113-29.
- 51 Harker LA, Hanson SR, Kelly AB. Antithrombotic benefits and hemorrhagic risks of direct thrombin antagonists. Thromb Haemost 1995; 74: 464-72.
- 52 Lee GF, Lazarus RA, Kelley RF. Potent bifunctional anticoagulants: Kunitz domain-tissue factor fusion proteins. Biochemistry 1997; 36: 5607-11.
- 53 Biesecker G, Dihel L, Enney K, Bendele RA. Derivation of RNA parameter inhibitors of human complement C5. Immunopharmacology 1999; 42: 219-30.
- 54 Ruckman J, Green LS, Beeson J, Waugh S, Gillette WL, Henninger DD, Claesson-Welsh L, Janjic N. 2’-Fluoropyrimidine RNA-based aptamers to the 165-amino acid form of vascular endothelial growth factor (VEGF165). Inhibition of receptor binding and VEGF-induced vascular permeability through interactions requiring the exon 7-encoded domain. J Biol Chem 1998; 273: 20556-67.
- 55 Willis MC, Collins BD, Zhang T, Green LS, Sebesta DP, Bell C, Kellogg E, Gill SC, Magallanez A, Knauer S, Bendele RA, Gill PS, Janjic N, Collins B. Liposome-achored vascular endothelial growth factor aptamers. Bioconjug Chem 1998; 09: 573-82.