Methods for inducing selective apoptosis
Provided herein are methods for cell therapy by modifying transfused cells to express an inducible caspase 9 protein, so that the cells may be selectively killed if the patient experiences dangerous side effects. Provided also within relates in part to methods for preventing or treating Graft versus Host Disease by modifying T cells before administration to a patient, so that they may be selectively killed if GvHD develops in the patient.
1. A method of administering transduced or transfected non-allodepleted allogeneic donor T cells to a human patient, comprising
a) administering transduced or transfected allogeneic donor T cells to the human patient, wherein:
the donor T cells have been transduced or transfected with a nucleic acid comprising
(i) a promoter region; and
(ii) a polynucleotide that encodes a chimeric protein comprising a multimeric ligand binding region and a caspase-9 polypeptide, wherein the promoter region is operatively linked to the polynucleotide and the multimeric ligand binding region comprises a FKBP12 polypeptide comprising a valine at position 36; and
the donor T cells are not allodepleted from the donor cell culture before transfection or transduction; and
b) administering a multimeric ligand that binds to the multimeric binding region to the patient, wherein:
the patient exhibits graft versus host disease symptoms wherein alloreactive donor T cells are present following a), and
the number of alloreactive donor T cells is reduced at least 90% within 24 hours following administration of the multimeric ligand.
2. The method of claim 1 , wherein the multimeric ligand is AP1903.
3. The method of claim 1 , wherein alloreactive T cells that are not undergoing cell division are ablated.
4. The method of claim 1 , wherein after administration of the multimeric ligand, donor T cells survive in the patient that are able to expand and are reactive to viruses and fungi.
5. The method of claim 1 , wherein after administration of the multimeric ligand, donor T cells survive in the patient that are able to expand and are reactive to tumor cells in the patient.
6. The method of claim 1 , further comprising administering a haploidentical stem cell transplant to the patient.
7. The method of claim 1 , wherein the chimeric protein further comprises a marker polypeptide.
8. The method of claim 7 , further comprising a selection step, wherein cells that express the marker are selected for administration to the patient.
9. The method of claim 8 , wherein the composition of the administered transduced or transfected T cells is over 90% pure.
10. The method of claim 1 , wherein the number of alloreactive donor T cells is reduced at least 90% within 30 minutes following administration of the multimeric ligand.
11. The method of claim 1 , wherein the donor T cells are not in an active state when administered to the patient.
12. The method of claim 1 , wherein the donor T cells are quiescent when administered to the patient.
13. The method of claim 1 , wherein the multimeric ligand binding region comprises two FKBP12 polypeptides, each comprising a valine substitution at position 36.
14. The method of claim 13 , wherein one of the FKBP12 polypeptides is encoded by a wobbled nucleotide sequence.
15. A method of treating graft versus host disease, comprising administering a multimeric ligand that binds to a multimeric ligand binding region to a human patient who has undergone allogeneic cell therapy using non-allodepleted donor T cells, wherein
a) allogeneic donor T cells introduced for the therapy express a chimeric protein comprising
(i) a multimeric ligand binding region comprising a FKBP polypeptide comprising a valine at position 36; and
(ii) a caspase-9 polypeptide;
b) following the allogeneic cell therapy, the patient exhibits graft versus host disease symptoms wherein alloreactive donor T cells are present; and
c) the number of alloreactive donor T cells is reduced at least 90% within 24 hours following administration of the multimeric ligand.
16. The method of claim 15 , wherein the caspase: 9 polypeptide is a truncated caspase: 9 polypeptide.
17. The method of claim 15 , further comprising administering a haploidentical stem cell transplant to the patient.
18. The method of claim 15 , wherein the chimeric protein further comprises a marker polypeptide.
19. The method of claim 15 , wherein the caspase: 9 polypeptide is a truncated caspase: 9 polypeptide.
20. The method of claim 15 , wherein the number of alloreactive donor T cells is reduced at least 90% within 30 minutes following administration of the multimeric ligand.
21. The method of claim 15 , wherein the multimeric ligand binding region comprises two FKBP12 polypeptides, each comprising a valine substitution at position 36.
22. The method of claim 21 , wherein one of the FKBP12 polypeptides is encoded by a wobbled nucleotide sequence.
23. The method of claim 15 , wherein the multimeric ligand is AP1903.
24. A method of administering transduced or transfected non-allodepleted allogeneic donor T cells to a human patient, comprising
a) administering transduced or transfected allogeneic donor T cells to the human patient, wherein:
the donor T cells have been transduced or transfected with a nucleic acid comprising
(i) a promoter region; and
(ii) a polynucleotide that encodes a chimeric protein comprising a multimeric ligand binding region and a caspase-9 polypeptide, wherein the promoter region is operatively linked to the polynucleotide and the muitimeric ligand binding region comprises a FKBP12 polypeptide comprising a valine at position 36; and
the donor T cells are not allodepleted from the donor cell culture before transfection or transduction; and
b) administering AP1903 to the patient, wherein:
the patient exhibits graft versus host disease symptoms wherein alloreactive donor T cells are present following a), and
the number of alloreactive donor T cells is reduced at least 90% within 24 hours following administration of AP1903.