Thromb Haemost 2012; 107(04): 717-725
DOI: 10.1160/TH11-11-0795
Platelets and Blood Cells
Schattauer GmbH

Disruption of PF4/H multimolecular complex formation with a minimally anticoagulant heparin (ODSH)

Manali V. Joglekar
1   Division of Hematology, Duke University Medical Center, Durham, North Carolina, USA
,
Pedro M. Quintana Diez
3   Paringenix, Inc, Weston, Florida, USA
,
Stephen Marcus
3   Paringenix, Inc, Weston, Florida, USA
,
Rui Qi
1   Division of Hematology, Duke University Medical Center, Durham, North Carolina, USA
,
Benjamin Espinasse
2   Department of Civil and Environmental Engineering, Duke University Medical Center, Durham, North Carolina, USA
,
Mark R. Wiesner
2   Department of Civil and Environmental Engineering, Duke University Medical Center, Durham, North Carolina, USA
,
Elizabeth Pempe
4   The University of North Carolina, School of Pharmacy and School of Medicine, Chapel Hill, North Carolina, USA
,
Jian Liu
4   The University of North Carolina, School of Pharmacy and School of Medicine, Chapel Hill, North Carolina, USA
,
Dougald M. Monroe
4   The University of North Carolina, School of Pharmacy and School of Medicine, Chapel Hill, North Carolina, USA
,
Gowthami M. Arepally
1   Division of Hematology, Duke University Medical Center, Durham, North Carolina, USA
› Institutsangaben
Weitere Informationen

Publikationsverlauf

Received: 16. November 2011

Accepted after minor revision: 24. Februar 2011

Publikationsdatum:
29. November 2017 (online)

Summary

Recent studies have shown that ultra-large complexes (ULCs) of platelet factor 4 (PF4) and heparin (H) play an essential role in the pathogenesis of heparin-induced thrombocytopenia (HIT), an immune-mediated disorder caused by PF4/H antibodies. Because antigenic PF4/H ULCs assemble through non-specific electrostatic interactions, we reasoned that disruption of charge-based interactions can modulate the immune response to antigen. We tested a minimally anticoagulant compound (2-O, 3-O desulfated heparin, ODSH) with preserved charge to disrupt PF4/H complex formation and immunogenicity. We show that ODSH disrupts complexes when added to pre-formed PF4/H ULCs and prevents ULC formation when incubated simultaneously with PF4 and UFH. In other studies, we show that excess ODSH reduces HIT antibody (Ab) binding in immunoassays and that PF4/ODSH complexes do not cross-react with HIT Abs. When ODSH and unfractionated heparin (UFH) are mixed at equimolar concentrations, we show that there is a negligible effect on amount of protamine required for heparin neutralisation and reduced immunogenicity of PF4/UFH in the presence of ODSH. Taken together, these studies suggest that ODSH can be used concurrently with UFH to disrupt PF4/H charge interactions and provides a novel strategy to reduce antibody mediated complications in HIT.

Presented in part at the 52nd American Society of Hematology Annual Meeting and Exposition, December 6th, 2010, Orlando, Florida, USA.

 
  • References

  • 1 Frederiksen JW. Cardiopulmonary bypass in humans: bypassing unfractionated heparin. Ann Thorac Surg 2000; 70: 1434-1443.
  • 2 Greinacher A. The use of direct thrombin inhibitors in cardiovascular surgery in patients with heparin-induced thrombocytopenia. Semin Thromb Hemost 2004; 30: 315-327.
  • 3 Kurup V. et al. Cardiac surgery in a patient with heparin-induced thrombocytopenia--cautions with use of the direct thrombin inhibitor, argatroban. Conn Med 2006; 70: 245-250.
  • 4 Nimjee SM. et al. The potential of aptamers as anticoagulants. Trends Cardiovasc Med 2005; 15: 41-45.
  • 5 Nimjee SM. et al. A novel antidote-controlled anticoagulant reduces thrombin generation and inflammation and improves cardiac function in cardiopulmonary bypass surgery. Mol Ther 2006; 14: 408-415.
  • 6 Arepally GM, Ortel TL. Heparin-Induced Thrombocytopenia. N Engl J Med 2006; 355: 809-817.
  • 7 Warkentin TE. et al. Anti-platelet factor 4/heparin antibodies in orthopedic surgery patients receiving antithrombotic prophylaxis with fondaparinux or enoxaparin. Blood 2005; 106: 3791-3796.
  • 8 Cuker A. Heparin-induced thrombocytopenia (HIT) in 2011: An epidemic of overdiagnosis. Thromb Haemost 2011; 106: 993-994.
  • 9 Lo GK. et al. What is the potential for overdiagnosis of heparin-induced throm-bocytopenia?. Am J Hematol 2007; 82: 1037-1043.
  • 10 Krauel K. et al. Heparin-induced thrombocytopenia--therapeutic concentrations of danaparoid, unlike fondaparinux and direct thrombin inhibitors, inhibit formation of platelet factor 4-heparin complexes. J Thromb Haemost 2008; 06: 2160-2167.
  • 11 Meuleman DG. Orgaran (Org 10172): its pharmacological profile in experimental models. Haemostasis 1992; 22: 58-65.
  • 12 Danhof M. et al. Pharmacokinetic considerations on Orgaran (Org 10172) therapy. Haemostasis 1992; 22: 73-84.
  • 13 Rao N V, Argyle B, Xu XY. et al. Low anticoagulant heparin targets multiple sites of inflammation, suppresses heparin-induced thrombocytopenia, and inhibits interaction of RAGE with its ligands. Am J Physiol Cell Physiol 2010; 299: C97-C110.
  • 14 Fryer A. et al. Selective O-desulfation produces nonanticoagulant heparin that retains pharmacological activity in the lung. J Pharmacol Exp Ther 1997; 282: 208-219.
  • 15 Atha DH. et al. Contribution of monosaccharide residues in heparin binding to antithrombin III. Biochemistry 1985; 24: 6723-6729.
  • 16 Arepally GM. et al. Characterization of a murine monoclonal antibody that mimics heparin-induced thrombocytopenia antibodies. Blood 2000; 95: 1533-1540.
  • 17 Suvarna S. et al. Determinants of PF4/heparin immunogeneicity. Blood 2007; 110: 4253-4260.
  • 18 Suvarna S. et al. PF4/heparin complexes are T cell-dependent antigens. Blood 2005; 106: 929-931.
  • 19 Rauova L. et al. Ultralarge complexes of PF4 and heparin are central to the pathogenesis of heparin-induced thrombocytopenia. Blood 2005; 105: 131-138.
  • 20 Saggin L. et al. Neutralization of the antiheparin activity of platelet factor 4 by a monoclonal antibody. Thromb Haemost 1992; 06: 137-143.
  • 21 Stringer SE, Gallagher JT. Specific binding of the chemokin platelet factor 4 to heparan sulfate. J Biol Chem 1997; 272: 20508-20514.
  • 22 Chudasama SL. et al. Heparin modifies the immunogenicity of positively-charged proteins. Blood 2010; 116: 6046-6053.
  • 23 Wilson WW. Light scattering as a diagnostic for protein crystal growth--a practical approach. J Struct Biol 2003; 142: 56-65.
  • 24 Suvarna S. et al. Optimization of a murine immunization model for study of PF4/heparin antibodies. J Thromb Haemost 2009; 07: 857-864.
  • 25 Hopfner M. et al. Selectin-blocking semisynthetic sulfated polysaccharides as promising anti-inflammatory agents. J Pharm Pharmacol 2003; 55: 697-706.
  • 26 Ferrer-Lopez P. et al. Heparin inhibits neutrophil-induced platelet activation via cathepsin G. J Lab Clin Med 1992; 119: 231-239.
  • 27 Krauel K. et al. Heparin-induced thrombocytopenia: in vitro studies on the interaction of dabigatran, rivaroxaban, and low-sulfated heparin, with platelet factor 4 and anti-PF4/heparin antibodies. Blood. 2011 epub ahead of print.
  • 28 Grocott H P. et al. Neurological injury during cardiopulmonary bypass in the rat. Perfusion 2001; 16: 75-81.
  • 29 Mackensen GB. et al. Cardiopulmonary bypass induces neurologic and neurocognitive dysfunction in the rat. Anesthesiology 2001; 95: 1485-1491.