Hyperacute rejection of the heart and kidney is reliably prevented when GalTKO organs are used that additionally express a human complement pathway regulatory protein (hCPRP). In contrast, although GalTKO.hCPRP pig lung xenograft function is significantly prolonged relative to GalTKO or hCPRP lungs, lung injury nonetheless occurs within hours of transplantation into baboons or following perfusion with human blood. Similarly, platelet sequestration occurs almost immediately when GalTKO.hCPRP pig livers are orthotopically transplanted in baboons, and limits recipient survival. A number of molecular incompatibilities between pig endothelial membrane glycoproteins and primate platelet receptors and plasma coagulation pathway proteins probably explain the mechanistic basis of coagulation pathway dysregulation and platelet sequestration in lung and liver xenograft injury. These incompatibilities may also contribute to related problems in heart and kidney xenotransplants, thrombotic microangiopathy (TM) and consumptive coagulopathy (CC). In this Project we will take advantage ofthe GalTKO.hCPRP pig lung's particular avidity for platelet and neutrophil sequestration, and its propensity to amplify coagulation pathway activation. Using a combination of clinically available approaches to impact thromboregulation (DDAVP, to deplete donor vWF pretransplant;zanamivir, a sialydase inhibitor, to maintain sialic acid expression on circulating platelet receptors and associated vWF) and unique molecular targeting reagents (an anti-GPlB Fab and an antisialydase antibody), Aim 1 will test the hypothesis that GPlB/vWF, modulated by desialydation, mediate primate platelet and neutrophil sequestration by porcine endothelium.
In Aim 2, GalTKO.hCPRP pig lungs that also express human thrombin pathway regulatory proteins (thrombomodulin or endothelial protein C receptor) will reveal whether molecular incompatibilities in this pathway contribute significantly to lung injury, and whether expression of these molecules is protective.
Aim 3 will determine whether strategies arising from Aims 1 and 2 will consistently yield life-supporting function of a pig lung in a baboon, and whether a similar approach reliably prevents thrombocytopenia in a baboon supported by a GalTKO.hCPRP pig liver.
This Project is likely to improve understanding of mechanisms of platelet sequestration, coagulation cascade activation, and organ xenograft injury that are probably relevant to lung and liver, and quite possibly to heart and kidney xenografts. By identifying a clinically applicable approach to prevent these phenomena, clinical application of pig liver xenografts as a bridge-to-allotransplant may result as well as progress to control TM and CC in Project 1.
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|Wijkstrom, Martin; Bottino, Rita; Iwase, Hayoto et al. (2015) Glucose metabolism in pigs expressing human genes under an insulin promoter. Xenotransplantation 22:70-9|
|Ezzelarab, Mohamed B; Ekser, Burcin; Azimzadeh, Agnes et al. (2015) Systemic inflammation in xenograft recipients precedes activation of coagulation. Xenotransplantation 22:32-47|
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|Iwase, Hayato; Ekser, Burcin; Hara, Hidetaka et al. (2014) Regulation of human platelet aggregation by genetically modified pig endothelial cells and thrombin inhibition. Xenotransplantation 21:72-83|
|Burdorf, L; Stoddard, T; Zhang, T et al. (2014) Expression of human CD46 modulates inflammation associated with GalTKO lung xenograft injury. Am J Transplant 14:1084-95|
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|Li, Jiang; Ezzelarab, Mohamed B; Ayares, David et al. (2014) The potential role of genetically-modified pig mesenchymal stromal cells in xenotransplantation. Stem Cell Rev 10:79-85|
|LaMattina, John C; Burdorf, Lars; Zhang, Tianshu et al. (2014) Pig-to-baboon liver xenoperfusion utilizing GalTKO.hCD46 pigs and glycoprotein Ib blockade. Xenotransplantation 21:274-86|
|Zhou, Huidong; Iwase, Hayato; Wolf, Roman F et al. (2014) Are there advantages in the use of specific pathogen-free baboons in pig organ xenotransplantation models? Xenotransplantation 21:287-90|
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