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 (aGT) 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 at least one complement inhibitor transgene and expression of at least one complement inhibitor transgene in at least pancreatic islet cells and tissues, wherein the at least one complement inhibitor transgene is selected from the group consisting of CD46, CD55 (DAF), CD59, CR1, and combinations thereof;
(iii) incorporation and expression of at least one immunosuppressive 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; and
(v) 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 of the transplanted porcine pancreatic cells into a host.
2. The porcine animal of claim 1 , wherein the phenotype of the porcine is characterized by specific expression of the at least one anticoagulant and the at least one immunosuppressant transgenes in pancreatic islet cells and tissues.
3. The porcine animal of claim 1 , wherein the at least one anticoagulant transgene is selected from the group consisting of tissue factor pathway inhibitor (TFPI), CD39, hirudin, thrombomodulin, endothelial cell protein C receptor (EPCR), and combinations thereof.
4. The porcine animal of claim 3 , wherein the anticoagulant transgene is tissue factor pathway inhibitor (TFPI).
5. The porcine animal of claim 3 , wherein the anti-coagulant transgene is CD39.
6. The porcine animal of claim 3 , wherein the anti-coagulant transgene is selected from the group consisting of hirudin, thrombomodulin, and endothelial cell protein C receptor (EPCR).
7. Pancreatic tissue derived from the porcine animal of claim 1 .
8. Pancreatic cells derived from the porcine animal of claim 1 .
9. The tissue of claim 7 , wherein the tissue is neonatal.
10. The cells of claim 8 , wherein the cells are pancreatic islet cells.
11. The cells of claim 10 , wherein the pancreatic islet cells are beta cells.
12. The cells of claim 10 , wherein the pancreatic islet cells are encapsulated.
13. The porcine animal of claim 1 , wherein the animal specifically expresses at least two anticoagulant transgenes in pancreatic tissue.
14. The porcine animal of claim 13 , wherein the anti-coagulant transgenes are TFPI and CD39.
15. 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 immunosuppressive transgene CTLA4.
16. The porcine animal of claim 1 , wherein the complement inhibitor is selected from the group consisting of CD46, CD55 (DAF) and combinations thereof.
17. The porcine animal of claim 16 , wherein the complement inhibitor is CD46.
18. Pancreatic tissue derived from the porcine animal of claim 16 .
19. Pancreatic cells derived from the porcine animal of claim 16 .
20. The porcine animal of claim 18 , wherein the tissue is neonatal.
21. The cells of claim 19 wherein the cells are pancreatic islets.
22. The cells of claim 19 , wherein the pancreatic islet cells are beta cells.
23. The cells of claim 19 , wherein the cells are encapsulated.
24. The porcine animal of claim 1 , wherein the immunosuppressive transgene is selected from the group consisting of CTLA4, CIITA, CD47, HLA-E, TRAIL, and combinations thereof.
25. Pancreatic tissue derived from the animal of claim 24 .
26. Pancreatic cells derived from the animal of claim 24 .
27. The tissue of claim 25 , wherein the tissue is neonatal.
28. The cells of claim 26 , wherein the cells are pancreatic islets.
29. The cells of claim 26 , wherein the pancreatic cells are beta cells.
30. The cells of claim 26 , wherein the cells are encapsulated.
31. The porcine animal of claim 24 , wherein the immunosuppressive transgene is CTLA4.