Development of a novel strategy to target CD39 antithrombotic activity to the endothelial-platelet microenvironment in kidney ischemia-reperfusion injury.
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.
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
M
Sashindranath
School of Medicine, Faculty of Health, Deakin University, Geelong, Australia.
Karen M
KM
Dwyer
Immunology Research Centre, St Vincent's Hospital, Melbourne, Australia.
Shala
S
Dezfouli
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.|Immunology Research Centre, St Vincent's Hospital, Melbourne, Australia.
Carly
C
Selan
Immunology Research Centre, St Vincent's Hospital, Melbourne, Australia.
Sandra
S
Crikis
Immunology Research Centre, St Vincent's Hospital, Melbourne, Australia.
Bo
B
Lu
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.
Yuping
Y
Yuan
Department of Medicine, Monash Medical Centre, Centre for Inflammatory Diseases, Monash University, Clayton, Australia.
Michael J
MJ
Hickey
Atherothrombosis and Vascular Laboratory, Baker Heart and Diabetes Institute and Monash University, Melbourne, Australia.
Karlheinz
K
Peter
Department of Medicine, Division of Gastroenterology, Harvard Medical School, Boston, USA.
Simon C
SC
Robson
Immunology Research Centre, St Vincent's Hospital, Melbourne, Australia.
Peter J
PJ
Cowan
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. harshal.nandurkar@monash.edu.
Harshal H
0000-0002-8767-116X
HH
Nandurkar
D000818:Animals; D015703:Antigens, CD / Q000494:pharmacology*; D001081:Apyrase / Q000494:pharmacology*; D001792:Blood Platelets / Q000187:drug effects; D060833:Cellular Microenvironment / Q000502:physiology; D004730:Endothelium, Vascular / Q000187:drug effects*; D005343:Fibrinolytic Agents / Q000494:pharmacology*; D006801:Humans; D007668:Kidney / Q000098:blood supply / Q000187:drug effects*; D008562:Membrane Glycoproteins / Q000494:pharmacology; D051379:Mice; D010974:Platelet Aggregation / Q000187:drug effects; D011994:Recombinant Proteins / Q000494:pharmacology; D015427:Reperfusion Injury
D015703:Antigens, CD; D005343:Fibrinolytic Agents; D008562:Membrane Glycoproteins; C084882:P-selectin ligand protein; D011994:Recombinant Proteins; D001081:Apyrase; C067869:CD39 antigen
CD39; Kidney ischemia–reperfusion injury; NTPDase-1; PSGL-1; Purinergic signalling