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.
Keywords
CD39 NTPDase-1 PSGL-1 Purinergic signalling Kidney ischemia–reperfusion injuryNotes
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).
Compliance with ethical standards
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.
Statement of competing financial interests
No competing financial interests.
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