Method for combined conditioning and chemoselection in a single cycle
A method of radiation-free hematopoietic stem cell (HSC) transplantation comprises administering to a mammalian subject one or two doses of 2 to 10 mg/kg body weight of a purine base analog, such as 6TG as a pre-conditioning step. The method further comprises engrafting into the subject hypoxanthine-guanine phosphoribosyltransferase (HPRT)-deficient donor HSCs within 48 to 72 hours of the pre-conditioning step; and administering to the subject about 1 to 5 mg/kg of the purine base analog every two to four days for two to eight weeks following the engrafting step. The method is performed in the absence of pre-conditioning via radiation. The subject is therefore not treated with myeloablative radiation in preparation for transplantation, and thus the subject is free of myeloablative radiation-induced toxicity.
1. A method of radiation-free hematopoietic stem cell (HSC) transplantation, the method comprising:
(a) administering to a mammalian subject one or two doses of 2 to 10 mg/kg body weight of a purine base analog selected from 6-thioguanine (6TG), 6-mercaptopurine (6-MP), or azathiopurine (AZA), as a pre-conditioning step;
(b) engrafting into the subject hypoxanthine-guanine phosphoribosyltransferase (HPRT)-deficient donor HSCs within 48 to 72 hours of the pre-conditioning step; and
(c) immediately administering to the subject about 1 to 5 mg/kg of the purine base analog every two to four days for two to eight weeks without allowing for a recovery period;
wherein the method is performed in the absence of pre-conditioning via radiation, and wherein the subject is suspected of having or has a disease or disorder.
2. The method of claim 1 , wherein the subject is human.
3. The method of claim 1 , wherein the subject has a hereditary or genetic disorder.
4. The method of claim 1 , wherein the subject has an acquired disease affecting lymphohematopoietic cells.
5. The method of claim 4 , wherein the disease or disorder is human immunodeficiency virus (HIV) infection or acquired immune deficiency syndrome (AIDS).
6. The method of claim 4 , wherein the disease or disorder is a lymphohematopoietic malignancy.
7. The method of claim 1 , wherein the disease or disorder is of hematopoietic or thrombopoietic or lymphopoietic system.
8. The method of claim 7 , wherein the disease or disorder of the hematopoietic system is a hemoglobinopathy.
9. The method of claim 1 , wherein the method further comprises bone marrow transplantation.
10. The method of claim 1 , wherein the purine base analog is 6TG.
11. The method of claim 1 , wherein the total 6TG dosage administered to the subject in the administering of steps (a) and (c) does not exceed 105 mg.
12. The method of claim 1 , wherein the total 6TG dosage administered to the subject in the administering of steps (a) and (c) does not exceed 75 mg.
13. The method of claim 1 , wherein the administering of step (c) is performed every 3 days and for not more than four weeks following the engrafting step.
14. The method of claim 1 , wherein no more than 5 days elapse between administering dosages of purine base analog throughout steps (a) to and (c).
15. The method of claim 1 , wherein the subject exhibits over 75% genetically modified hematopoietic cells.
16. The method of claim 1 , wherein the subject exhibits over 95% genetically modified hematopoietic cells.
17. The method of claim 1 , wherein the HPRT-deficient HSCs to be transplanted have been rendered HPRT-deficient via introduction of sequences encoding zinc finger nucleases (ZFNs), transcriptional activator-like effector nucleases (TALENs), small fragment homologous recombination (SFHR) template strands, inhibitory RNAs (siRNAs) or microRNAs (miRNAs), antisense RNAs, trans-splicing RNAs, ribozymes, intracellular antibodies, or dominant-negative or competitive inhibitor proteins.
18. The method of claim 1 , wherein the HPRT-deficient HSCs to be transplanted have been rendered HPRT-deficient via introduction of sequences encoding zinc finger nucleases (ZFNs).
19. The method of claim 1 , wherein the HPRT-deficient HSCs to be transplanted have been rendered HPRT-deficient via introduction of sequences encoding transcriptional activator-like effector nucleases (TALENs).
20. The method of claim 1 , wherein the HPRT-deficient HSCs to be transplanted have been rendered HPRT-deficient via introduction of sequences encoding inhibitory RNAs (siRNAs).
21. The method of claim 1 , wherein the HPRT-deficient HSCs to be transplanted have been genetically modified.
22. The method of claim 1 , wherein the transplanted HSCs are autologous or syngeneic.
23. The method of claim 1 , wherein the transplanted HSCs are allogeneic.
24. The method of claim 1 , wherein the subject is not treated with myeloablative radiation.
25. A method of treating symptoms of a disease or disorder affecting lymphohematopoietic cells in a subject, the method comprising:
(a) administering to the subject one or two doses of 2 to 10 mg/kg body weight of a purine base analog selected from 6-thioguanine (6TG), 6-mercaptopurine (6-MP), or azathiopurine (AZA), as a pre-conditioning step;
(b) engrafting into the subject hypoxanthine-guanine phosphoribosyltransferase (HPRT)-deficient donor HSCs within 48 to 72 hours of the pre-conditioning step; and
(c) immediately administering to the subject about 1 to 5 mg/kg of the purine base analog every two to four days for two to eight weeks without allowing for a recovery period;
wherein the method is performed in the absence of pre-conditioning via radiation.
26. The method of claim 25 , wherein the disease or disorder is human immunodeficiency virus (HIV) infection or acquired immune deficiency syndrome (AIDS).
27. The method of claim 25 , wherein the disease or disorder is a lymphohematopoietic malignancy.
28. The method of claim 25 , wherein the disease or disorder is of hematopoietic or thrombopoietic or lymphopoietic system.
29. The method of claim 28 , wherein the disease or disorder of the hematopoietic system is a hemoglobinopathy.
30. The method of claim 25 , wherein the method further comprises bone marrow or hematopoietic stem cell transplantation.
31. The method of claim 25 , wherein the subject is human.
32. The method of claim 25 , wherein the purine base analog is 6TG.
33. The method of claim 25 , wherein the administering of step (c) is performed every 3 days and for not more than four weeks following the engrafting step.
34. The method of claim 25 , wherein no more than 5 days elapse between administering dosages of purine base analog throughout steps (a) to and (c).
35. The method of claim 25 , wherein the subject exhibits over 75% genetically modified hematopoietic cells.
36. The method of claim 25 , wherein the subject exhibits over 95% genetically modified hematopoietic cells.
37. The method of claim 25 , wherein the HPRT-deficient HSCs to be transplanted have been rendered HPRT-deficient via introduction of sequences encoding zinc finger nucleases (ZFNs), transcriptional activator-like effector nucleases (TALENs), small fragment homologous recombination (SFHR) template strands, inhibitory RNAs (siRNAs) or microRNAs (miRNAs), antisense RNAs, trans-splicing RNAs, ribozymes, intracellular antibodies, or dominant-negative or competitive inhibitor proteins.
38. The method of claim 25 , wherein the HPRT-deficient HSCs to be transplanted have been rendered HPRT-deficient via introduction of sequences encoding zinc finger nucleases (ZFNs).
39. The method of claim 25 , wherein the HPRT-deficient HSCs to be transplanted have been rendered HPRT-deficient via introduction of sequences encoding transcriptional activator-like effector nucleases (TALENs).
40. The method of claim 25 , wherein the HPRT-deficient HSCs to be transplanted have been rendered HPRT-deficient via introduction of sequences encoding inhibitory RNAs (siRNAs).
41. The method of claim 25 , wherein the HPRT-deficient HSCs to be transplanted have been genetically modified.
42. The method of claim 25 , wherein the transplanted HSCs are autologous or syngeneic.
43. The method of claim 25 , wherein the transplanted HSCs are allogeneic.
44. The method of claim 25 , wherein the subject is not treated with myeloablative radiation.