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Thromb Haemost 2010; 103(02): 426-435
DOI: 10.1160/TH09-05-0305
Cardiovascular Biology and Cell Signalling
Schattauer GmbH

Human solCD39 inhibits injury-induced development of neointimal hyperplasia

Joan H. F. Drosopoulos
1   Thrombosis Research Laboratory, Research Service, VA New York Harbor Healthcare System, New York, New York, USA
2   Division of Hematology and Medical Oncology, Department of Medicine, Weill Cornell Medical College, New York, New York, USA
,
Rosemary Kraemer
3   Division of Hematology and Medical Oncology, Department of Pathology and Laboratory Medicine, Weill Cornell Medical College, New York, New York, USA
,
Hao Shen
3   Division of Hematology and Medical Oncology, Department of Pathology and Laboratory Medicine, Weill Cornell Medical College, New York, New York, USA
,
Rita K. Upmacis
3   Division of Hematology and Medical Oncology, Department of Pathology and Laboratory Medicine, Weill Cornell Medical College, New York, New York, USA
,
Aaron J. Marcus
1   Thrombosis Research Laboratory, Research Service, VA New York Harbor Healthcare System, New York, New York, USA
2   Division of Hematology and Medical Oncology, Department of Medicine, Weill Cornell Medical College, New York, New York, USA
3   Division of Hematology and Medical Oncology, Department of Pathology and Laboratory Medicine, Weill Cornell Medical College, New York, New York, USA
,
Elgilda Musi
1   Thrombosis Research Laboratory, Research Service, VA New York Harbor Healthcare System, New York, New York, USA
› Author Affiliations Financial support: This work was supported by a Department of Veterans Affairs Merit Review grant and NIH grants HL046403 and HL047073.
Further Information

Publication History

Received: 13 May 2009

Accepted after major revision: 01 November 2009

Publication Date:
22 November 2017 (online)

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Summary

Blood platelets provide the initial response to vascular endothelial injury, becoming activated as they adhere to the injured site. Activated platelets recruit leukocytes, and initiate proliferation and migration of vascular smooth muscle cells (SMC) within the injured vessel wall, leading to development of neointimal hyperplasia. Endothelial CD39/NTPDase1 and recombinant solCD39 rapidly metabolise nucleotides, including stimulatory ADP released from activated platelets, thereby suppressing additional platelet reactivity. Using a murine model of vascular endothelial injury, we investigated whether circulating human solCD39 could reduce platelet activation and accumulation, thus abating leukocyte infiltration and neointimal formation following vascular damage. Intraperitoneally-administered solCD39 ADP -ase activity in plasma peaked 1 hour (h) post-injection, with an elimination half-life of 43 h. Accordingly, mice were administered solCD39 or saline 1 h prior to vessel injury, then either sacrificed 24 h post-injury or treated with solCD39 or saline (three times weekly) for an additional 18 days. Twenty-four hours post-injury, solCD39-treated mice displayed a reduction in platelet activation and recruitment, P-selectin expression, and leukocyte accumulation in the arterial lumen. Furthermore, repeated administration of solCD39 modulated the late stage of vascular injury by suppressing leukocyte deposition, macrophage infiltration and smooth muscle cell (SMC) proliferation/migration, resulting in abrogation of neointimal thickening. In contrast, injured femoral arteries of saline-injected mice exhibited massive platelet thrombus formation, marked P-selectin expression, and leukocyte infiltration. Pronounced neointimal growth with macrophage and SMC accretion was also observed (intimal-to-medial area ratio 1.56 ± 0.34 at 19 days). Thus, systemic administration of solCD39 profoundly affects injury-induced cellular responses, minimising platelet deposition and leukocyte recruitment, and suppressing neointimal hyperplasia.

Keywords

CD39 - endothelial E-NTPDase1 - arterial injury - platelet activation - vascular stenosis
 

  • References

  • 1 Roque M, Fallon JT, Badimon JJ. et al. Mouse model of femoral artery denudation injury associated with the rapid accumulation of adhesion molecules on the luminal surface and recruitment of neutrophils. Arterioscler Thromb Vasc Biol 2000; 20: 335-342.
    CrossrefPubMedGoogle Scholar
  • 2 Mitra AK, Gangahar DM, Agrawal DK. Cellular, molecular and immunological mechanisms in the pathophysiology of vein graft intimal hyperplasia. Immunol Cell Biol 2006; 84: 115-124.
    CrossrefPubMedGoogle Scholar
  • 3 May AE, Langer H, Seizer P. et al. Platelet-leukocyte interactions in inflammation and atherothrombosis. Semin Thromb Hemost 2007; 33: 123-127.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 4 Siegel-Axel DI, Gawaz M. Platelets and endothelial cells. Semin Thromb Hemost 2007; 33: 128-135.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 5 van Gils JM, Zwaginga JJ, Hordijk PL. Molecular and functional interactions among monocytes, platelets, and endothelial cells and their relevance for cardiovascular diseases. J Leukoc Biol 2009; 85: 195-204.
    CrossrefPubMedGoogle Scholar
  • 6 Jennings LK. Mechanisms of platelet activation: Need for new strategies to protect against platelet-mediated atherothrombosis. Thromb Haemost 2009; 102: 248-257.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 7 Seye CI, Kong Q, Yu N. et al. P2 receptors in atherosclerosis and postangioplasty restenosis. Purinergic Signal 2007; 3: 153-162.
    CrossrefPubMedGoogle Scholar
  • 8 Erlinge D, Burnstock G. P2 receptors in cardiovascular regulation and disease. Purinergic Signal 2008; 4: 1-20.
    PubMedGoogle Scholar
  • 9 Marcus AJ, Broekman MJ, Drosopoulos JH. et al. Heterologous cell-cell interactions: thromboregulation, cerebroprotection and cardioprotection by CD39 (NTPDase-1). J Thromb Haemost 2003; 1: 2497-2509.
    CrossrefPubMedGoogle Scholar
  • 10 Marcus AJ, Broekman MJ, Drosopoulos JHF. et al. The endothelial cell ecto-ADPase responsible for inhibition of platelet function is CD39. J Clin Invest 1997; 99: 1351-1360.
    CrossrefPubMedGoogle Scholar
  • 11 Kaczmarek E, Koziak K, Sévigny J. et al. Identification and characterization of CD39 vascular ATP diphosphohydrolase. J Biol Chem 1996; 271: 33116-33122.
    CrossrefPubMedGoogle Scholar
  • 12 Maliszewski CR, Delespesse GJ, Schoenborn MA. et al. The CD39 lymphoid cell activation antigen. Molecular cloning and structural characterization. J Immunol 1994; 153: 3574-3583.
    PubMedGoogle Scholar
  • 13 Koziak K, Sévigny J, Robson SC. et al. Analysis of CD39/ATP diphosphohydrolase (ATPDase) expression in endothelial cells, platelets and leukocytes. Thromb Haemost 1999; 82: 1538-1544.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 14 Sevigny J, Sundberg C, Braun N. et al. Differential catalytic properties and vascular topography of murine nucleoside triphosphate diphosphohydrolase 1 (NTPDase1) and NTPDase2 have implications for thromboregulation. Blood 2002; 99: 2801-2809.
    CrossrefPubMedGoogle Scholar
  • 15 Glenn JR, White AE, Johnson A. et al. Leukocyte count and leukocyte ecto-nucleotidase are major determinants of the effects of adenosine triphosphate and adenosine diphosphate on platelet aggregation in human blood. Platelets 2005; 16: 159-170.
    CrossrefPubMedGoogle Scholar
  • 16 Pulte ED, Broekman MJ, Olson KE. et al. CD39/NTPDase-1 activity and expression in normal leukocytes. Thromb Res 2007; 121: 309-317.
    CrossrefPubMedGoogle Scholar
  • 17 Zimmermann H. Ectonucleotidases: Some recent developments and a note on nomenclature. Drug Dev Res 2001; 52: 44-56.
    CrossrefPubMedGoogle Scholar
  • 18 Wang TF, Ou Y, Guidotti G. The transmembrane domains of ectoapyrase (CD39) affect its enzymatic activity and quaternary structure. J Biol Chem 1998; 273: 24814-24821.
    CrossrefPubMedGoogle Scholar
  • 19 Schulte am Esch J, Sévigny J, Kaczmarek E. et al. Structural elements and limited proteolysis of CD39 influence ATP diphosphohydrolase activity. Biochemistry 1999; 38: 2248-2258.
    CrossrefPubMedGoogle Scholar
  • 20 Gayle RB, Maliszewski CR, Gimpel SD. et al. Inhibition of platelet function by recombinant soluble ecto-ADPase/CD39. J Clin Invest 1998; 101: 1851-1859.
    CrossrefPubMedGoogle Scholar
  • 21 Drosopoulos JHF, Broekman MJ, Islam N. et al. Site-directed mutagenesis of human endothelial cell ecto-ADPase/soluble CD39. Requirement of gluta-mate174 and serine218 for enzyme activity and inhibition of platelet recruitment. Biochemistry 2000; 39: 6936-6943.
    CrossrefPubMedGoogle Scholar
  • 22 Drosopoulos JHF. Roles of Asp54 and Asp213 in Ca2+ utilization by soluble human CD39/ecto-nucleotidase. Arch Biochem Biophys 2002; 406: 85-95.
    CrossrefPubMedGoogle Scholar
  • 23 Pinsky DJ, Broekman MJ, Peschon JJ. et al. Elucidation of the thromboregulatory role of CD39/ectoapyrase in the ischemic brain. J Clin Invest 2002; 109: 1031-1040.
    CrossrefPubMedGoogle Scholar
  • 24 Buergler JM, Maliszewski CR, Broekman MJ. et al. Effects of SolCD39, a novel inhibitor of platelet aggregation, on platelet deposition and aggregation after PTCA in a porcine model. J Thromb Thrombolysis 2005; 19: 115-122.
    CrossrefPubMedGoogle Scholar
  • 25 Belayev L, Khoutorova L, Deisher TA. et al. Neuroprotective effect of SolCD39, a novel platelet aggregation inhibitor, on transient middle cerebral artery occlusion in rats. Stroke 2003; 34: 758-763.
    CrossrefPubMedGoogle Scholar
  • 26 Musi E, Islam N, Drosopoulos JHF. Constraints imposed by transmembrane domains affect enzymatic activity of membrane-associated human CD39/NTPDase1 mutants. Arch Biochem Biophys 2007; 461: 30-39.
    CrossrefPubMedGoogle Scholar
  • 27 Smyth SS, Reis ED, Zhang W. et al. β3-Integrin-deficient mice but not P-selectin-deficient mice develop intimal hyperplasia after vascular injury: Correlation with leukocyte recruitment to adherent platelets 1 hour after injury. Circulation 2001; 103: 2501-2507.
    CrossrefPubMedGoogle Scholar
  • 28 Kanda K, Hayman GT, Silverman MD. et al. Comparison of ICAM-1 and VCAM-1 expression in various human endothelial cell types and smooth muscle cells. Endothelium 1998; 6: 33-44.
    CrossrefPubMedGoogle Scholar
  • 29 Muller WA. Leukocyte-endothelial cell interactions in the inflammatory response. Lab Invest 2002; 82: 521-533.
    CrossrefPubMedGoogle Scholar
  • 30 Woodfin A, Voisin MB, Nourshargh S. PECAM-1: a multi-functional molecule in inflammation and vascular biology. Arterioscler Thromb Vasc Biol 2007; 27: 2514-2523.
    CrossrefPubMedGoogle Scholar
  • 31 Voisard R, Osswald M, Baur R. et al. Expression of intercellular adhesion molecule-1 in human coronary endothelial and smooth muscle cells after stimulation with tumor necrosis factor-alpha. Coron Artery Dis 1998; 9: 737-745.
    CrossrefPubMedGoogle Scholar
  • 32 Manka DR, Wiegman P, Din S. et al. Arterial injury increases expression of inflammatory adhesion molecules in the carotid arteries of apolipoprotein-E-deficient mice. J Vasc Res 1999; 36: 372-378.
    CrossrefPubMedGoogle Scholar
  • 33 O’Brien KD, Allen MD, McDonald TO. et al. Vascular cell adhesion molecule-1 is expressed in human coronary atherosclerotic plaques. Implications for the mode of progression of advanced coronary atherosclerosis. J Clin Invest 1993; 92: 945-951.
    CrossrefPubMedGoogle Scholar
  • 34 Osaka M, Hagita S, Haraguchi M. et al. Real-time imaging of mechanically injured femoral artery in mice reveals a biphasic pattern of leukocyte accumulation. Am J Physiol 2007; 292: H1876-H1882.
    PubMedGoogle Scholar
  • 35 Kikuchi S, Umemura K, Kondo K. et al. Photochemically induced endothelial injury in the mouse as a screening model for inhibitors of vascular intimal thickening. Arterioscler Thromb Vasc Biol 1998; 18: 1069-1078.
    CrossrefPubMedGoogle Scholar
  • 36 Tanaka H, Sukhova GK, Swanson SJ. et al. Sustained activation of vascular cells and leukocytes in the rabbit aorta after balloon injury. Circulation 1993; 88: 1788-1803.
    CrossrefPubMedGoogle Scholar
  • 37 Wang K, Zhou X, Zhou Z. et al. Platelet, not endothelial, P-selectin is required for neointimal formation after vascular injury. Arterioscler Thromb Vasc Biol 2005; 25: 1584-1589.
    CrossrefPubMedGoogle Scholar
  • 38 Koyama H, Maeno T, Fukumoto S. et al. Platelet P-selectin expression is associated with atherosclerotic wall thickness in carotid artery in humans. Circulation 2003; 108: 524-529.
    CrossrefPubMedGoogle Scholar
  • 39 Fateh-Moghadam S, Li Z, Ersel S. et al. Platelet degranulation is associated with progression of intima-media thickness of the common carotid artery in patients with diabetes mellitus type 2. Arterioscler Thromb Vasc Biol 2005; 25: 1299-1303.
    CrossrefPubMedGoogle Scholar
  • 40 Zernecke A, Bidzhekov K, Ozuyaman B. et al. CD73/ecto-5‘-nucleotidase protects against vascular inflammation and neointima formation. Circulation 2006; 113: 2120-2127.
    CrossrefPubMedGoogle Scholar
  • 41 Elliott MR, Chekeni FB, Trampont PC. et al. Nucleotides released by apoptotic cells act as a find-me signal to promote phagocytic clearance. Nature 2009; 461: 282-286.
    CrossrefPubMedGoogle Scholar
  • 42 Kukulski F, Levesque SA, Lavoie EG. et al. Comparative hydrolysis of P2 receptor agonists by NTPDases 1, 2, 3 and 8. Purinergic Signal 2005; 1: 193-204.
    CrossrefPubMedGoogle Scholar
  • 43 Koziak K, Bojakowska M, Robson SC. et al. Overexpression of CD39/nucleoside triphosphate diphosphohydrolase-1 decreases smooth muscle cell proliferation and prevents neointima formation after angioplasty1. J Thromb Haemost 2008; 6: 1191-1197.
    CrossrefPubMedGoogle Scholar
  • 44 Imai M, Takigami K, Guckelberger O. et al. Recombinant adenoviral mediated CD39 gene transfer prolongs cardiac xenograft survival. Transplantation 2000; 70: 864-870.
    CrossrefPubMedGoogle Scholar
  • 45 Gangadharan SP, Imai M, Rhynhart KK. et al. Targeting platelet aggregation: CD39 gene transfer augments nucleoside triphosphate diphosphohydrolase activity in injured rabbit arteries. Surgery 2001; 130: 296-303.
    CrossrefPubMedGoogle Scholar
  • 46 Dwyer KM, Robson SC, Nandurkar HH. et al. Thromboregulatory manifestations in human CD39 transgenic mice and the implications for thrombotic disease and transplantation. J Clin Invest 2004; 113: 1440-1446.
    CrossrefPubMedGoogle Scholar
  • 47 Haller CA, Cui W, Wen J. et al. Reconstitution of CD39 in liposomes amplifies nucleoside triphosphate diphosphohydrolase activity and restores thromboregulatory properties. J Vasc Surg 2006; 43: 816-823.
    CrossrefPubMedGoogle Scholar