Multi-transgenic pigs for diabetes treatment
The present invention provides certain animals, and in particular porcine animals, tissue and cells derived from these, which lack any expression of functional alpha 1,3 galactosyltransferase (αGT) and express one or more additional transgenes which make them suitable donors for pancreatic islet xenotransplantation. Methods of treatment and prevention of diabetes using cells derived from such animals are also provided.
1. A genetically modified transgenic porcine animal, wherein the genetic modifications comprise modifications to the genome of the porcine that result in (i) the lack of any expression of functional alpha 1,3 galactosyltransferase (GTKO), (ii) incorporation of a human CD46 transgene under the control of the human CD46 promoter and expression of the human CD46 transgene in at least pancreatic islet cells and tissues; (iii) incorporation and expression of at least one immunosuppressant transgene under the control of an islet-specific promoter in the genome of pancreatic islet cells and tissues; (iv) incorporation and expression of at least one anticoagulant transgene under the control of an islet-specific promoter in the genome of pancreatic islet cells and tissues; (v) wherein the phenotype of the porcine is characterized by specific expression of the anticoagulant and immunosuppressant transgenes in pancreatic islet cells and tissues; and (vi) wherein transgenic porcine islet cells isolated from the porcine animal produce insulin and reduce the instant blood mediated inflammatory reaction (IBMIR), in comparison to non-transgenic porcine islet cells, after transplantation into a host.
2. The porcine animal of claim 1 , wherein the anticoagulant is selected from the group consisting of tissue factor pathway inhibitor (TFPI), CD39, hirudin, thrombomodulin and endothelial cell protein C receptor (EPCR).
3. The porcine animal of claim 2 , wherein the anticoagulant is tissue factor pathway inhibitor (TFPI).
4. The porcine animal of claim 2 , wherein the anti-coagulant is CD39.
5. The porcine animal of claim 2 , wherein the anti-coagulant is selected from the group consisting of hirudin, thrombomodulin and endothelial cell protein C receptor (EPCR).
6. Pancreatic tissue derived from the porcine animal of claim 1 .
7. Pancreatic cells derived from the porcine animal of claim 1 .
8. The tissue of claim 6 , wherein the tissue is neonatal.
9. The cells of claim 7 , wherein the cells are pancreatic islets.
10. The cells of claim 7 , wherein the pancreatic cells are beta cells.
11. The cells of claim 7 , wherein the cells are encapsulated.
12. The porcine animal of claim 1 , wherein the animal specifically expresses at least two anticoagulant transgenes in pancreatic tissue.
13. The porcine animal of claim 12 , wherein the anti-coagulants are TFPI and CD39.
14. The porcine animal of claim 1 , wherein the animal specifically expresses at least three transgenes in pancreatic tissue wherein the at least three transgenes are the anticoagulant transgenes TFPI and CD39 and the immunosuppressant transgene CTLA4.
15. Pancreatic tissue derived from the animal of claim 14 .
16. Pancreatic cells derived from the animal of claim 14 .
17. The tissue of claim 15 , wherein the tissue is neonatal.
18. The cells of claim 16 wherein the cells are pancreatic islets.
19. The cells of claim 16 , wherein the pancreatic cells are beta cells.
20. The cells of claim 16 , wherein the cells are encapsulated.
21. The porcine animal of claim 1 , wherein the immunosuppressant is selected from the group consisting of CTLA4, CIITA, CD47, HLA-E, and TRAIL.
22. The porcine animal of claim 1 , wherein the immunosuppressant is CTLA4.
23. Pancreatic tissue derived from the animal of claim 21 .
24. Pancreatic cells derived from the animal of claim 21 .
25. The tissue of claim 23 , wherein the tissue is neonatal.
26. The cells of claim 24 wherein the cells are pancreatic islets.
27. The cells of claim 24 , wherein the pancreatic cells are beta cells.
28. The cells of claim 24 , wherein the cells are encapsulated.