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Development of a novel strategy to target CD39 antithrombotic activity to the endothelial-platelet microenvironment in kidney ischemia–reperfusion injury

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Abstract

Kidney ischemia–reperfusion injury (IRI) is common during transplantation. IRI is characterised by inflammation and thrombosis and associated with acute and chronic graft dysfunction. P-selectin and its ligand PSGL-1 are cell adhesion molecules that control leukocyte-endothelial and leukocyte-platelet interactions under inflammatory conditions. CD39 is the dominant vascular nucleotidase that facilitates adenosine generation via extracellular ATP/ADP-phosphohydrolysis. Adenosine signalling is protective in renal IRI, but CD39 catalytic activity is lost with exposure to oxidant stress. We designed a P-selectin targeted CD39 molecule (rsol.CD39-PSGL-1) consisting of recombinant soluble CD39 that incorporates 20 residues of PSGL-1 that bind P-selectin. We hypothesised that rsol.CD39-PSGL-1 would maintain endothelial integrity by focusing the ectonucleotidase platelet-inhibitory activity and reducing leukocyte adhesion at the injury site. The rsol.CD39-PSGL-1 displayed ADPase activity and inhibited platelet aggregation ex vivo, as well as bound with high specificity to soluble P-selectin and platelet surface P-selectin. Importantly, mice injected with rsol.CD39-PSGL-1 and subjected to renal IRI showed significantly less kidney damage both biochemically and histologically, compared to those injected with solCD39. Furthermore, the equivalent dose of rsol.CD39-PSGL-1 had no effect on tail template bleeding times. Hence, targeting recombinant CD39 to the injured vessel wall via PSGL-1 binding resulted in substantial preservation of renal function and morphology after IRI without toxicity. These studies indicate potential translational importance to clinical transplantation and nephrology.

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References

  1. Crikis S et al (2010) Transgenic overexpression of CD39 protects against renal ischemia-reperfusion and transplant vascular injury. Am J Transplant 10(12):2586–2595

    Article  CAS  PubMed  Google Scholar 

  2. Lu B et al (2008) The impact of purinergic signaling on renal ischemia-reperfusion injury. Transplantation 86(12):1707–1712

    Article  CAS  PubMed  Google Scholar 

  3. Rajakumar SV et al (2010) Deficiency or inhibition of CD73 protects in mild kidney ischemia-reperfusion injury. Transplantation 90(12):1260–1264

    Article  CAS  PubMed  Google Scholar 

  4. Hohmann JD et al (2013) Delayed targeting of CD39 to activated platelet GPIIb/IIIa via a single-chain antibody: breaking the link between antithrombotic potency and bleeding? Blood 121(16):3067–3075

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Roberts V et al (2013) The CD39-adenosinergic axis in the pathogenesis of renal ischemia-reperfusion injury. Purinergic Signal 9(2):135–143

    Article  CAS  PubMed  Google Scholar 

  6. Kaczmarek E et al (1996) Identification and characterization of CD39/vascular ATP diphosphohydrolase. J Biol Chem 271(51):33116–33122

    Article  CAS  PubMed  Google Scholar 

  7. Zimmermann H (1992) 5'-Nucleotidase: molecular structure and functional aspects. Biochem J 285(Pt 2):345–365

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Pinsky DJ et al (2002) Elucidation of the thromboregulatory role of CD39/ectoapyrase in the ischemic brain. JClinInvest 109(8):1031–1040

    CAS  Google Scholar 

  9. Imai M et al (1999) Modulation of nucleoside [correction of nucleotide] triphosphate diphosphohydrolase-1 (NTPDase-1)cd39 in xenograft rejection. Mol Med 5(11):743–752

    CAS  PubMed  PubMed Central  Google Scholar 

  10. Imai M et al (2000) Recombinant adenoviral mediated CD39 gene transfer prolongs cardiac xenograft survival. Transplantation 70(6):864–870

    Article  CAS  PubMed  Google Scholar 

  11. Belayev L et al (2003) Neuroprotective effect of SolCD39, a novel platelet aggregation inhibitor, on transient middle cerebral artery occlusion in rats. Stroke 34(3):758–763

    Article  CAS  PubMed  Google Scholar 

  12. Gayle RB 3rd et al (1998) Inhibition of platelet function by recombinant soluble ecto-ADPase/CD39. J Clin Invest 101(9):1851–1859

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. McEver RP, Cummings RD (1997) Perspectives series: cell adhesion in vascular biology. Role of PSGL-1 binding to selectins in leukocyte recruitment. J Clin Invest 100(3):485–491

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Yang J, Furie BC, Furie B (1999) The biology of P-selectin glycoprotein ligand-1: its role as a selectin counterreceptor in leukocyte-endothelial and leukocyte-platelet interaction. Thromb Haemost 81(1):1–7

    CAS  PubMed  Google Scholar 

  15. Vandendries ER, Furie BC, Furie B (2004) Role of P-selectin and PSGL-1 in coagulation and thrombosis. Thromb Haemost 92(3):459–466

    CAS  PubMed  Google Scholar 

  16. Burch EE et al (2002) The N-terminal peptide of PSGL-1 can mediate adhesion to trauma-activated endothelium via P-selectin in vivo. Blood 100(2):531–538

    Article  CAS  PubMed  Google Scholar 

  17. Liu W et al (1998) Identification of N-terminal residues on P-selectin glycoprotein ligand-1 required for binding to P-selectin. J Biol Chem 273(12):7078–7087

    Article  CAS  PubMed  Google Scholar 

  18. Li F et al (1996) Post-translational modifications of recombinant P-selectin glycoprotein ligand-1 required for binding to P- and E-selectin. J Biol Chem 271(6):3255–3264

    Article  CAS  PubMed  Google Scholar 

  19. Straub A et al (2011) Evidence of platelet activation at medically used hypothermia and mechanistic data indicating ADP as a key mediator and therapeutic target. Arterioscler Thromb Vasc Biol 31(7):1607–1616

    Article  CAS  PubMed  Google Scholar 

  20. Mountford JK et al (2015) The class II PI 3-kinase, PI3KC2alpha, links platelet internal membrane structure to shear-dependent adhesive function. Nat Commun 6:6535

    Article  CAS  PubMed  Google Scholar 

  21. Dejana E et al (1979) Bleeding time in laboratory animals. III—do tail bleeding times in rats only measure a platelet defect? (The aspirin puzzle). Thromb Res 15(1–2):199–207

    Article  CAS  PubMed  Google Scholar 

  22. Swers JS et al (2006) A high affinity human antibody antagonist of P-selectin mediated rolling. Biochem Biophys Res Commun 350(3):508–513

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported by grants awarded to HH Nandurkar from the NHMRC (344801) and NIH (1 R01 HL078651) and to SC Robson from the NIH (5 R01 HL094400).

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Authors and Affiliations

  1. Australian Centre for Blood Diseases, Central Clinical School, Alfred Hospital, Monash University, Monash AMREP building, Level 1, Walkway, via The Alfred Centre, 99 Commercial Road, Melbourne, VIC, 3004, Australia

    Maithili Sashindranath, Carly Selan, Yuping Yuan & Harshal H. Nandurkar

  2. School of Medicine, Faculty of Health, Deakin University, Geelong, Australia

    Karen M. Dwyer

  3. Immunology Research Centre, St Vincent’s Hospital, Melbourne, Australia

    Shala Dezfouli, Carly Selan, Sandra Crikis, Bo Lu & Peter J. Cowan

  4. Department of Medicine, Monash Medical Centre, Centre for Inflammatory Diseases, Monash University, Clayton, Australia

    Michael J. Hickey

  5. Atherothrombosis and Vascular Laboratory, Baker Heart and Diabetes Institute and Monash University, Melbourne, Australia

    Karlheinz Peter

  6. Department of Medicine, Division of Gastroenterology, Harvard Medical School, Boston, USA

    Simon C. Robson

Authors
  1. Maithili Sashindranath
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  2. Karen M. Dwyer
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  3. Shala Dezfouli
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  4. Carly Selan
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  5. Sandra Crikis
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  6. Bo Lu
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  7. Yuping Yuan
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  8. Michael J. Hickey
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  9. Karlheinz Peter
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  10. Simon C. Robson
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  11. Peter J. Cowan
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  12. Harshal H. Nandurkar
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Corresponding author

Correspondence to Harshal H. Nandurkar.

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Conflict of interest

Maithili Sashindranath declares that she has no conflict of interest.

Karen M Dwyer declares that she has no conflict of interest.

Shala Dezfouli declares that she has no conflict of interest.

Carly Selan declares that she has no conflict of interest.

Sandra Crikis declares that she has no conflict of interest.

Bo Lu declares that he has no conflict of interest.

Yuping Yuan declares that she has no conflict of interest.

Michael J Hickey declares that he has no conflict of interest.

Karlheinz Peter declares that he has no conflict of interest.

Simon C Robson declares that he has no conflict of interest.

Peter J Cowan declares that he has no conflict of interest.

Harshal Nandurkar declares that he has no conflict of interest.

Ethical approval

This article does not contain any studies with human participants . All animal experiments were approved by the St Vincent’s Hospital Animal Ethics Committee and were conducted in compliance with the Australian code for the care and use of animals for scientific purposes.

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No competing financial interests.

Additional information

Peter J Cowan and Harshal H Nandurkar are equal senior authors.

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Sashindranath, M., Dwyer, K.M., Dezfouli, S. et al. Development of a novel strategy to target CD39 antithrombotic activity to the endothelial-platelet microenvironment in kidney ischemia–reperfusion injury. Purinergic Signalling 13, 259–265 (2017). https://doi.org/10.1007/s11302-017-9558-3

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