Therapeutic alteration of transplantable tissues through in situ or ex vivo exposure to RNA interference molecules
The present invention, at least in part, relates to the discovery of efficacious delivery of an RNAi agent (in preferred aspects of the invention, an siRNA) to a transplantable tissue. Organ rejection, transplantation-mediated transmission of viral infection, and triggering of apoptosis in transplanted tissues can each be minimized by the methods and compositions of the instant invention. The RNAi agent(s) of the instant invention can be delivered as “naked” molecules, or using liposomal and other modes of delivery, to transplantable tissues. Such delivery can occur via perfusion of the RNAi agent in solution through the vasculature of a whole or partial organ; or tissues including transplantable cells and cell lines may be bathed, injected or otherwise treated with RNAi agents. Preferred transplantable tissues include, for example, pancreas, liver, kidney, heart, lung, and all cells and cell lines derived from such tissues (e.g., pancreatic islet cells that may, e.g., be transplanted as a treated population).
1. A method of treating a subject at risk of or having a disorder associated with apoptosis-induced ischemic injury in an organ, the method comprising administering to the subject an RNAi agent capable of downmodulating ischemia-induced apoptosis, wherein the RNAi agent comprises a sense strand and an anti-sense strand, wherein the anti-sense strand is sufficiently complementary to p53 RNA to reduce the production of p53 via RNA-induced silencing complex (RISC) so as to counter the apoptosis-induced ischemic injury in the organ, and wherein said administering comprises injecting the RNAi agent into the organ.
2. The method of claim 1 , wherein the method is directed to apoptosis-induced ischemic injury in at least one of the group consisting of: a kidney, liver, pancreas, heart, and lung.
3. The method of claim 1 , wherein the RNAi agent is in a preservation solution that comprises either saline or an immunosuppressant.
4. The method of claim 1 , wherein the RNAi agent is an siRNA molecule.
5. The method of claim 1 , wherein the RNAi agent is an shRNA molecule.
6. The method of claim 1 , wherein the RNAi agent comprises an oligonucleotide comprising a modification to a 2′-OH group, the modification being selected from the group consisting of the 2′-OH group replaced by a H, alkoxy or OR, halogen, SH, SR, amino, or CN group, wherein R is a lower alkyl, alkenyl, alkynyl, or aryl.
7. The method of claim 6 , wherein the RNAi agent comprises a 2′-0-methyl oligonucleotide.
8. The method of claim 1 , wherein the RNAi agent contacts the organ via lipid-mediated delivery or as a component of a liposomal delivery preparation.
9. A method of treating a subject at risk of or having a disorder associated with apoptosis-induced ischemic injury, the method comprising administering to the subject by perfusion an RNAi agent capable of downmodulating ischemia-induced apoptosis, wherein the RNAi agent comprises a sense strand and an anti-sense strand, wherein the anti-sense strand is sufficiently complementary to p53 RNA to reduce production of p53 via RNA-induced silencing complex (RISC), so as to counter apoptosis-induced ischemic injury.
10. The method of claim 9 , wherein the apoptosis is associated with transplantation of an organ.
11. The method of claim 9 , wherein the apoptosis is not associated with transplantation of an organ.
12. A method of treating a subject at risk of or having a disorder or condition associated with apoptosis-induced ischemic injury, the method comprising administering to the subject by perfusion an RNAi agent comprising a sense strand and an anti-sense guide strand, wherein the guide strand is sufficiently complementary to p53 RNA to downregulate the production of p53 by effecting RNA-induced silencing complex (RISC)-mediated cleavage of the p53 mRNA, inhibiting expression of the p53 RNA so as to counter or reduce the disorder or condition.
13. The method of claim 9 , wherein the method is directed to apoptosis-induced ischemic injury in at least one of the group consisting of: a kidney, liver, pancreas, heart, lung, and islet cells.
14. The method of claim 12 , wherein the method is directed to apoptosis-induced ischemic injury in at least one of the group consisting of: a kidney, liver, pancreas, heart, lung, and islet cells.