Inhibition of coagulation proteases Xa and IIa decreases ischemia-reperfusion injuries in a preclinical renal transplantation model

• Published in: Translational research : the journal of laboratory and clinical medicine Vol. 178; pp. 95 - 106.e1
• Main Authors: Tillet, S, Giraud, S, Kerforne, T, Saint Yves, T, Joffrion, S, Goujon, J.M, Cau, J, Mauco, G, Petitou, M, Hauet, T
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.12.2016; Elsevier
• Subjects: Animals; Biochemistry, Molecular Biology; Biomarkers - metabolism; Biotin - analogs & derivatives; Biotin - pharmacology; Blood Coagulation - drug effects; Cold Temperature; Endothelial Cells - metabolism; Epithelial-Mesenchymal Transition - drug effects; Factor Xa - metabolism; Factor Xa Inhibitors; Fibrosis; Human health and pathology; Humans; Hypoxia - complications; Inflammation - pathology; Internal Medicine; Kidney - drug effects; Kidney Function Tests; Kidney Transplantation; Leukocytes - drug effects; Leukocytes - pathology; Life Sciences; Models, Animal; Oligosaccharides - pharmacology; Prothrombin - antagonists & inhibitors; Prothrombin - metabolism; Reperfusion Injury - enzymology; Reperfusion Injury - pathology; Sus scrofa; Thrombin - metabolism; Urology and Nephrology; PAR; Protease Activated Receptors; Monocyte Chemoattractant Molecule-1; Primary Human Aortic Endothelial Cells; TAT; clone anti swine monocytes/granulocytes; ICAM-1; Donors Deceased after Cardiac Death; uninephrectomised animals; Tumor Necrosis Factor alpha; α-SMA; Organ preservation; University of Wisconsin; Plasminogen Activator Inhibitor type-1; FBS; SWC3a; Tissue Factor; iNOS; PAI-1; National Institute for Agronomic Research; Transforming Growth Factor-β; Epithelial–Mesenchymal Transition; UW; IL; Vascular Cell Adhesion Molecule-1; DCD; Fetal Bovine Serum; MMP2; Intercellular Adhesion Molecule-1; IR; Kidney Transplantation; Nitric Oxide; EMT; INRA; Matrix MetalloProteinase-2; Ischemia-Reperfusion; Ischemia-Reperfusion Injuries; GAPDH; GlycerAldehyde 3-Phosphate DeHydrogenase; Hematoxylin - Eosin - Safran; IRI; NO; MCP-1; Interleukin; Thrombin-AntiThrombin complexes; Anticoagulant; UnFractionated Heparin; UFH; TGF-β; α-Smooth Muscle Actin; TF; VCAM-1; TNF-α; HAEC; HES; NEP; inductible Nitric Oxide Synthase; IRIs; MMP-2
• ISSN: 1931-5244; 1878-1810
• DOI: 10.1016/j.trsl.2016.07.014

Abstract Coagulation is an important pathway in the pathophysiology of ischemia-reperfusion injuries. In particular, deceased after circulatory death donors undergo a no-flow period, a strong activator of coagulation. Hence, therapies influencing the coagulation cascade must be developed. We evaluated the effect of a new highly specific and effective anti-Xa/IIa molecule, with an integrated innovative antidote site (EP217609), in a porcine preclinical model mimicking injuries observed in deceased after circulatory death donor kidney transplantation. Kidneys were clamped during 60 min (warm ischemia), then flushed and preserved 24h in 4°C University of Wisconsin (UW) solution (supplemented or not). EP217609 was used in UW (UW-EP), compared to unfractionated-heparin (UW-UFH) or UW alone (UW). A mechanistic investigation was conducted in vitro : addition of EP217609 on endothelial cells during hypoxia at 4°C in UW inhibited thrombin generation at the 37°C reoxygenation in human plasma and reduced TNF-α, ICAM-1 and VCAM-1 mRNA cells expression. In vivo , function recovery was markedly improved in UW-EP group. Interestingly, levels of thrombin-antithrombin complexes (reflecting thrombin generation) were reduced 60 min after reperfusion in the UW-EP group. Three months after transplantation, lower fibrosis, epithelial-mesenchymal-transition, inflammation and leukocytes infiltration were observed in the UW-EP group. Using this new dual anticoagulant anti-Xa/IIa activity during kidney flush and preservation is protective by reducing thrombin generation at revascularization, improving early function recovery and decreasing chronic lesions. Such an easy-to-deploy clinical strategy could improve marginal graft outcome.