ERYTHROID CELLS COMPRISING ARGININE DEIMINASE
Compositions comprising synthetic membrane-receiver complexes, methods of generating synthetic membrane-receiver complexes, and methods of treating or preventing diseases, disorders or conditions therewith.
1 . An enucleated erythroid cell comprising an exogenous polypeptide comprising arginine deiminase or a functional fragment thereof,
wherein the enucleated erythroid cell is made by a process comprising introducing into an erythroid cell precursor a nucleic acid encoding the exogenous polypeptide.
2 . The enucleated erythroid cell of claim 1 , which comprises at least 1,000 copies of the exogenous polypeptide.
3 . The enucleated erythroid cell of claim 1 , which comprises at least 10,000 copies of the exogenous polypeptide.
4 . The enucleated erythroid cell of claim 1 , wherein the exogenous polypeptide is intracellular.
5 . The enucleated erythroid cell of claim 1 , wherein the exogenous polypeptide is on the surface of the enucleated erythroid cell.
6 . The enucleated erythroid cell of claim 1 , wherein the exogenous polypeptide is not fused to an endogenous polypeptide.
7 . The enucleated erythroid cell of claim 1 , wherein the exogenous polypeptide consists essentially of arginine deiminase.
8 . The enucleated erythroid cell of claim 1 , wherein the exogenous polypeptide consists of arginine deiminase.
9 . The enucleated erythroid cell of claim 1 , further comprising a second exogenous polypeptide comprising an arginine transporter.
10 . The enucleated erythroid cell of claim 1 , which exhibits an increase in arginine deiminase activity of at least 2-fold relative to that of an enucleated erythroid cell that does not comprise the exogenous polypeptide.
11 . The enucleated erythroid cell of claim 1 , which is a reticulocyte.
12 . The enucleated erythroid cell of claim 1 , which is an erythrocyte.
13 . The enucleated erythroid cell of claim 1 , which lacks A and B antigens.
14 . The enucleated erythroid cell of claim 1 , which is a human cell.
15 . The enucleated erythroid cell of claim 1 , which comprises fetal hemoglobin.
16 . The enucleated erythroid cell of claim 1 , which exhibits substantially the same osmotic membrane fragility as an isolated, unmodified, uncultured enucleated erythroid cell.
17 . The enucleated erythroid cell of claim 1 , wherein introducing the nucleic acid comprises using a lentiviral vector.
18 . The enucleated erythroid cell of claim 1 , wherein the process comprises expanding the erythroid cell precursor by at least 20,000-fold in culture.
19 . The enucleated erythroid cell of claim 1 , wherein the nucleated erythroid cell precursor is a CD34+ hematopoietic stem cell.
20 . The enucleated erythroid cell of claim 1 , wherein the nucleic acid comprises DNA.
21 . The enucleated erythroid cell of claim 1 , wherein the nucleic acid comprises RNA.
22 . A pharmaceutical composition comprising a plurality of the enucleated erythroid cells of claim 1 and a pharmaceutically acceptable carrier.
23 . The pharmaceutical composition of claim 22 , which is formulated for intravenous administration.
24 . The pharmaceutical composition of claim 22 , wherein at least about 90% of enucleated erythroid cells in the pharmaceutical composition comprise the exogenous polypeptide.
25 . A pharmaceutical composition comprising (i) a plurality of the enucleated erythroid cells of claim 1 , wherein at least 70% of cells in the pharmaceutical composition are enucleated, and (ii) a pharmaceutically acceptable carrier.
26 . The pharmaceutical composition of claim 25 , wherein at least 90% of cells in the pharmaceutical composition are enucleated.
27 . A nucleated erythroid cell precursor comprising an exogenous polypeptide comprising arginine deiminase or a functional fragment thereof,
wherein the nucleated erythroid cell precursor was made by a process comprising introducing an exogenous nucleic acid encoding the exogenous polypeptide into the nucleated erythroid cell precursor.
28 . The nucleated erythroid cell precursor of claim 27 , which has been cultured after the introduction of the exogenous nucleic acid.
29 . A method of reducing arginine levels in a subject, comprising administering to the subject the pharmaceutical composition of claim 22 , thereby reducing arginine levels in the subject.
30 . A method of treating hepatocellular carcinoma or melanoma in a subject in need thereof, the method comprising administering to the subject the pharmaceutical composition of claim 22 , thereby treating said hepatocellular carcinoma or melanoma in the subject.