Genetically modified stem cells
Described herein are human transgenic beta cells expressing fugetactic levels of CXCL12 to a subject in need thereof. Also described herein are beta cells comprising a transgene comprising a nucleic acid sequence encoding CXCL12.
1. A population of isolated genetically engineered human beta cells comprising a nucleic acid encoding for exogenous or heterologous expression of a human CXCL12 protein isoform such that said population of said cells is characterized as encoding for the expression of a fugetactic amount of said CXCL12 protein isoform making said cell population resistant to cell death when in the presence of human immune cells in vitro as compared to a population of unengineered human beta cells,
wherein said population of genetically engineered human beta cells and said population of genetically unengineered human beta cells are obtained from a human stem cell that has been differentiated ex vivo into human beta cells,
wherein the nucleic acid is operably linked to an exogenous promoter,
wherein the nucleic acid encodes a human CXCL12 protein isoform that has an amino acid sequence identity to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, or SEQ ID NO:6,
wherein the population of genetically engineered human beta cells produces insulin, and
wherein said population of genetically engineered human beta cells and said population of genetically unengineered human beta cells is either autologous or allogeneic.
2. The population of isolated human beta cells of claim 1 , wherein the population of genetically engineered human beta cells is allogeneic.
3. The population of isolated human beta cells of claim 1 ,
wherein the fugetactic amount is determined by determining the amount of lactate dehydrogenase (LDH) produced by said population of genetically engineered human beta cells cultured for 48 hours in the presence of human peripheral blood mononuclear cells (PBMCs) at a ratio of 1:30 genetically engineered human beta cells to human PBMCs; and
wherein the amount of LDH produced by said population of engineered human beta cells is at least 50% less than the amount of LDH produced by a population of unengineered human beta cells, and at least 95% of the engineered human beta cells in the population of engineered cells survive after 48 hours of exposure to human PBMCs.
4. The population of isolated beta cells of claim 1 , wherein said beta cells comprise the alpha isoform or SEQ ID NO: 1.
5. The population of isolated beta cells of claim 1 , wherein said beta cells comprise a beta isoform or SEQ ID NO: 2.
6. The population of isolated beta cells of claim 1 , wherein said beta cells comprise (a) a gamma, delta, epsilon, or phi isoform, or (b) SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5 or SEQ ID NO: 6.
7. The population of isolated beta cells of claim 1 , wherein said human immune cells comprise B-cells, NK cells, and/or T-cells.
8. The population of isolated beta cells of claim 7 , wherein said T-cells are cytotoxic T-cells.
9. A pharmaceutical composition comprising a population of isolated beta cells of claim 1 and a pharmaceutically acceptable aqueous carrier.
10. The pharmaceutical composition of claim 9 , wherein said composition is suitable for injection.