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. An allogenic, genetically engineered, isolated human stem cell comprising an expression vector, wherein the expression vector comprises a nucleic acid sequence encoding a human CXCL12 protein, wherein the nucleic acid is operably linked to an exogenous regulatory region, and wherein said human stem cell, upon differentiation into beta cells, expresses a fugetactic amount of CXCL12 protein in said beta cell microenvironment.
2. The genetically engineered isolated human stem cell of claim 1 , wherein the human stem cell expresses a fugetactic amount of at least about 100 nM to 900 nM of CXCL12 protein in the cell microenvironment upon differentiation of the stem cell into beta cells obtained or derived from a subject with type 1 diabetes.
3. The genetically engineered isolated human stem cell of claim 1 , wherein the human stem cell is obtained or derived from a subject free of type 1 diabetes.
4. The genetically engineered isolated human stem cell of claim 1 , wherein the CXCL12 protein is selected from SDF-1alpha, SDF-1beta, SDF-1gamma, SDF-1delta, SDF-1epsilon, and SDF-1phi.
5. The genetically engineered isolated human stem cell of claim 4 , wherein the CXCL12 protein is selected from SDF-1alpha, SDF-1beta, SDF-1gamma, and SDF-1delta.
6. The genetically engineered isolated human stem cell of claim 1 , wherein the nucleic acid encoding for human CXCL12 protein comprises the nucleic acid sequence of SEQ ID NO.: 8 or SEQ ID NO.: 9.
7. The genetically engineered isolated human stem cell of claim 6 , wherein the nucleic acid encoding for human CXCL12 protein comprises the nucleic acid sequence of SEQ ID NO.: 8.
8. The genetically engineered isolated human stem cell of claim 6 , wherein the nucleic acid encoding for human CXCL12 protein comprises the nucleic acid sequence of SEQ ID NO.: 9.
9. The genetically engineered isolated human stem cell of claim 1 , wherein the human CXCL12 protein comprises the amino acid sequence of 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.
10. The genetically engineered isolated human stem cell of claim 1 , wherein the isolated human stem cell is a multipotent progenitor cell, an adult stem cell, a pluripotent stem cell, or an induced pluripotent stem cell.
11. The genetically engineered isolated human stem cell of claim 1 , wherein the exogenous regulatory region comprises a heterologous promoter.
12. The genetically engineered isolated human stem cell of claim 11 , wherein the heterologous promoter is an exogenous constitutive or inducible promoter.
13. The genetically engineered isolated human stem cell of claim 1 , wherein the nucleic acid encoding for human CXCL12 protein operably linked to the exogenous regulatory region was introduced into the stem cell by genome editing.
14. A population of the genetically engineered isolated human stem cells of claim 1 .
15. The allogeneic stem cell of claim 1 , wherein the differentiated beta cell expressing a fugetactic amount of CXCL12 has the characteristic that when present in vitro to human peripheral blood mononuclear cells (PBMCs) at a ratio of 1:30 genetically engineered beta cells to PBMCs for 24 hours decreases the amount of lipase dehydrogenase (LDH) by at least 95% as compared to a human beta cell produced from said allogeneic stem cell that is not engineered to express CXCL12.