PRODUCTION AND USE OF RED BLOOD CELLS
The present disclosure relates to the production of red blood cells from hematopoietic stem cells, by differentiating such cells in the presence of a protein that induces cell survival and proliferation.
1 .- 65 . (canceled)
66 . A method for treating a disease or disorder characterized by a deficiency of anucleated red blood cells in a subject in need thereof comprising:
(a) culturing a population of hematopoietic stem cells in a differentiation media comprising erythropoietin (EPO) and a recombinant fusion protein comprising a MYC polypeptide fused to a protein transduction domain, wherein culturing the hematopoietic stem cells induces differentiation of the hematopoietic stem cells to mature red blood cells, thereby producing a population of mature red blood cells; and
(b) administering to the subject the population of mature red blood cells.
67 . The method of claim 66 , wherein the recombinant fusion protein is TAT-MYC.
68 . The method of claim 66 , wherein the differentiation media further comprises one or more recombinant proteins that inhibit apoptosis, wherein the one or more recombinant proteins that inhibit apoptosis comprises one or more Bcl2 homology domains selected from the group consisting of BH1, BH2, BH3, and BH4.
69 . The method of claim 66 , wherein the hematopoietic stem cells further comprise one or more recombinant proteins of interest.
70 . The method of claim 66 , wherein the hematopoietic stem cells comprise one or more transgenes that encode one or more recombinant proteins of interest.
71 . The method of claim 70 , wherein the expression or function of the one or more recombinant proteins of interest is controllable or inducible.
72 . The method of claim 66 , wherein the disease or disorder characterized by a deficiency of anucleated red blood cells is selected from one or more of anemia, Gaucher's disease, hemolysis, neutropenia, thrombocytopenia, granulocytopenia, hemophilia, Hodgkin's lymphoma, Non-Hodgkin's lymphoma, B cell chronic lymphoma, Burkitt's lymphoma, Follicular-like lymphoma, diffused large B-cell lymphoma, multiple myeloma, acute myeloid leukemia, pre-B acute lymphocytic leukemia, pre-T acute lymphocytic leukemia, acute promyelocytic leukemia, or refractory leukemia.
73 . The method of claim 72 , wherein the disease or disorder characterized by a deficiency of anucleated red blood cells is anemia.
74 . The method of claim 66 , wherein the disease or disorder characterized by a deficiency of anucleated red blood cells results from one or more of chemotherapy, chemical exposure, radiation therapy, and/or radiation exposure.
75 . The method of claim 66 , wherein the population of mature red blood cells is co-administered with one or more of chemotherapy, radiation therapy, or one or more proteins of interest.
76 . The method of claim 66 , wherein the disease or disorder characterized by a deficiency of anucleated red blood cells comprises an infectious disease associated with hemorrhage.
77 . The method of claim 76 , wherein the infectious disease associated with hemorrhage comprises a viral hemorrhagic fever.
78 . The method of claim 77 , wherein the viral hemorrhagic fever is selected from Lassa fever, Ebola, Marburg hemorrhagic fever, Rift Valley fever, dengue fever, or yellow fever.
79 . The method of claim 66 , wherein the population of mature red blood cells exhibits one or more characteristics selected from the group consisting of: a population of mature red blood cells, wherein about 40% to about 100% of the cells are anucleated; a population of mature red blood cells, wherein about 40% to about 100% of the cells express glycophorin A (GPA); a population of mature red blood cells, wherein about 40% to about 100% of the cells express adult hemoglobin; a population of mature red blood cells, wherein about 40% to about 100% of the cells exhibit decreased levels of CD71 expression; and a population of mature red blood cells, wherein about 40% to about 100% of the cells exhibit decreased levels of fetal hemoglobin expression.
80 . The method of claim 66 , wherein the population of hematopoietic stem cells is derived from bone marrow, peripheral blood, mobilized peripheral blood, umbilical cord, placenta or fetal tissue, or is a hematopoietic cell line.
81 . The method of claim 66 , wherein the subject is human.