Erythroid cells comprising arginine deiminase

Abstract

Compositions comprising synthetic membrane-receiver complexes, methods of generating synthetic membrane-receiver complexes, and methods of treating or preventing diseases, disorders or conditions therewith.

What is claimed is: 
     
         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.