Hematopoietic precursor cell production by programming
The invention generally regards methods for providing hematopoietic cells and precursors of hematopoietic cells from a variety of cell sources, such as pluripotent stem cells or somatic cells. Also provided are therapeutic compositions including the provided hematopoietic cells and precursors of hematopoietic cells, and methods of using such for the treatment of subjects.
1. An in vitro method of providing human hematopoietic precursor cells by forward programming of human induced pluripotent stem cells, the method comprising:
providing the hematopoietic precursor cells by culturing the induced pluripotent stem cells under conditions to increase the expression level of one or more hematopoietic precursor programming factor gene capable of causing forward programming of the induced pluripotent stem cells into hematopoietic precursor cells, thereby forward programming the induced pluripotent stem cells into hematopoietic precursor cells, wherein the culturing is performed in the absence of bone marrow stromal cells, wherein the one or more hematopoietic precursor programming factor genes is under the control of a heterologous promoter, wherein the at least one hematopoietic precursor programming factor gene comprises an endothelial differentiation factor gene selected from the group consisting of ERG (v-ets erythroblastosis virus E26 oncogene homolog (avian)), FLI-1 (Friend leukemia virus integration 1), and ETV2 (ets variant 2); or comprises a hematopoietic precursor programming factor gene selected from the group consisting of GFI1 (growth factor independent 1 transcription repressor), GFI1B (growth factor independent 1B transcription repressor), TAL1 (T-cell acute lymphocytic leukemia), LYL1 (lymphoblastic leukemia derived sequence 1), LMO2 (LIM domain only 2 (rhombotin-like 1)), GATA2 (GATA binding protein 2), GATA3 (GATA binding protein 3), and SPI1 (spleen focus forming virus (SFFP) proviral integration oncogene spi1).
2. The method of claim 1 , wherein the hematopoietic precursor cells are provided by forward programming of induced pluripotent stem cells.
3. The method of claim 1 , wherein at least one hematopoietic precursor programming factor gene comprises an endothelial differentiation factor gene.
4. The method of claim 3 , wherein the at least one endothelial differentiation factor comprises ERG (v-ets erythroblastosis virus E26 oncogene homolog (avian)), FLI-1 (Friend leukemia virus integration 1), or ETV2 (ets variant 2).
5. The method of claim 4 , wherein at least one endothelial differentiation factor is ERG.
6. The method of claim 5 , wherein the ERG is ERG-2 or ERG-3.
7. The method of claim 1 , wherein the heterologous promoter is an inducible promoter.