Pig-to-baboon liver xenoperfusion utilizing GalTKO.hCD46 pigs and glycoprotein Ib blockade

Xenotransplantation. 2014 May-Jun;21(3):274-86. doi: 10.1111/xen.12093. Epub 2014 Mar 17.

Abstract

Background: Although transplantation of genetically modified porcine livers into baboons has yielded recipient survival for up to 7 days, survival is limited by profound thrombocytopenia, which becomes manifest almost immediately after revascularization, and by subsequent coagulopathy. Porcine von Willebrand's factor (VWF), a glycoprotein that adheres to activated platelets to initiate thrombus formation, has been shown to constitutively activate human platelets via their glycoprotein Ib (GPIb) receptors. Here, we report our pig-to-primate liver xenoperfusion model and evaluate whether targeting the GPIb-VWF axis prevents platelet sequestration.

Methods: Twelve baboons underwent cross-circulation with the following extracorporeal livers: one allogeneic control with a baboon liver, 4 xenogeneic controls with a GalTKO.hCD46 pig liver, 3 GalTKO.hCD46 pig livers in recipients treated with αGPIb antibody during perfusion, and 4 GalTKO.hCD46 pig livers pre-treated with D-arginine vasopressin (DDAVP) in recipients treated with αGPIb antibody during perfusion.

Results: All perfused livers appeared grossly and macroscopically normal and produced bile. Xenograft liver perfusion experiments treated with αGPIb antibody may show less platelet sequestration during the initial 2 h of perfusion. Portal venous resistance remained constant in all perfusion experiments. Platelet activation studies demonstrated platelet activation in all xenoperfusions, but not in the allogeneic perfusion.

Conclusion: These observations suggest that primate platelet sequestration by porcine liver and the associated thrombocytopenia are multifactorial and perhaps partially mediated by a constitutive interaction between porcine VWF and the primate GPIb receptor. Control of platelet sequestration and consumptive coagulopathy in liver xenotransplantation will likely require a multifaceted approach in our clinically relevant perfusion model.

Keywords: ex vivo perfusion; liver transplantation; xenotransplantation.

Publication types

  • Evaluation Study
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Genetically Modified
  • Biomarkers / metabolism
  • Extracorporeal Circulation
  • Galactosyltransferases / genetics
  • Gene Knockout Techniques
  • Graft Survival
  • Humans
  • Immunoglobulin Fab Fragments / therapeutic use*
  • Immunosuppressive Agents / therapeutic use*
  • Liver Transplantation / methods*
  • Membrane Cofactor Protein / genetics
  • Papio
  • Platelet Glycoprotein GPIb-IX Complex / immunology*
  • Platelet Glycoprotein GPIb-IX Complex / metabolism
  • Postoperative Complications / etiology
  • Postoperative Complications / prevention & control*
  • Swine / genetics
  • Thrombocytopenia / etiology
  • Thrombocytopenia / prevention & control*
  • Transplantation, Heterologous / methods*
  • von Willebrand Factor / metabolism

Substances

  • Biomarkers
  • CD46 protein, human
  • Immunoglobulin Fab Fragments
  • Immunosuppressive Agents
  • Membrane Cofactor Protein
  • Platelet Glycoprotein GPIb-IX Complex
  • von Willebrand Factor
  • Galactosyltransferases
  • alpha-1,3-galactosyltransferase 1, porcine