Elucidation of the thromboregulatory role of CD39/ectoapyrase in the ischemic brain
David J. Pinsky, … , Aaron J. Marcus, Charles R. Maliszewski
David J. Pinsky, … , Aaron J. Marcus, Charles R. Maliszewski
Published April 15, 2002
Citation Information: J Clin Invest. 2002;109(8):1031-1040. https://doi.org/10.1172/JCI10649.
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Article Vascular biology

Elucidation of the thromboregulatory role of CD39/ectoapyrase in the ischemic brain

Abstract

Endothelial CD39 metabolizes ADP released from activated platelets. Recombinant soluble human CD39 (solCD39) potently inhibited ex vivo platelet aggregation in response to ADP and reduced cerebral infarct volumes in mice following transient middle cerebral artery occlusion, even when given 3 hours after stroke. Postischemic platelet and fibrin deposition were decreased and perfusion increased without increasing intracerebral hemorrhage. In contrast, aspirin did not increase postischemic blood flow or reduce infarction volume, but did increase intracerebral hemorrhage. Mice lacking the enzymatically active extracellular portion of the CD39 molecule were generated by replacement of exons 4–6 (apyrase-conserved regions 2–4) with a PGKneo cassette. Although CD39 mRNA 3′ of the neomycin cassette insertion site was detected, brains from these mice lacked both apyrase activity and CD39 immunoreactivity. Although their baseline phenotype, hematological profiles, and bleeding times were normal, cd39–/– mice exhibited increased cerebral infarct volumes and reduced postischemic perfusion. solCD39 reconstituted these mice, restoring postischemic cerebral perfusion and rescuing them from cerebral injury. These data demonstrate that CD39 exerts a protective thromboregulatory function in stroke.

Authors

David J. Pinsky, M. Johan Broekman, Jacques J. Peschon, Kim L. Stocking, Tomoyuki Fujita, Ravichandran Ramasamy, E. Sander Connolly Jr., Judy Huang, Szilard Kiss, Yuan Zhang, Tanvir F. Choudhri, Ryan A. McTaggart, Hui Liao, Joan H.F. Drosopoulos, Virginia L. Price, Aaron J. Marcus, Charles R. Maliszewski

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Comparative effects of vehicle, aspirin, and solCD39 on the various outc...
Comparative effects of vehicle, aspirin, and solCD39 on the various outcomes of experimental stroke in cd39+/+ mice. (a) Relative cerebral blood flows shown at occlusion, reperfusion, and sacrifice at 23 hours after reperfusion following stroke for four treatment groups: 4 mg/kg solCD39 given preoperatively (n = 16) or 3 hours postoperatively (n = 9), aspirin given postoperatively (n = 6), and control (vehicle, given postoperatively; n = 10). (b) Relative cerebral blood flow at 24 hours in mice treated preoperatively with vehicle (n = 24), aspirin (n = 27), or solCD39 (n = 11, 11, and 16 for the 1 mg/kg, 2 mg/kg, and 4 mg/kg doses, respectively). Cerebral blood flow data for vehicle or solCD39, shown in a, are repeated here for comparison. At 24 hours, the following parameters were also determined: (c) cerebral infarct volume; (d) neurological deficit score, with higher scores indicating worse deficit (15); (e) mortality; and (f) intracerebral hemorrhage. *P < 0.05; **P < 0.01; ***P < 0.001; †P < 0.0001; ††P = 0.00002.