Synthetic membrane-receiver complexes
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 at least 1,000 copies of an exogenous polypeptide comprising phenylalanine hydroxylase (PAH), or a functional fragment thereof, wherein the enucleated erythroid cell was produced by a process comprising:
providing a nucleated erythroid cell, or a precursor thereof, comprising an exogenous nucleic acid encoding the exogenous polypeptide; and
culturing the nucleated erythroid cell under conditions suitable for enucleation of the nucleated erythroid cell and for production of the exogenous polypeptide.
2. The enucleated erythroid cell of claim 1 , wherein the enucleated erythroid cell exhibits substantially the same osmotic membrane fragility as a corresponding isolated, unmodified, uncultured erythroid cell.
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 , which comprises at least 50,000 copies of the exogenous polypeptide.
5. The enucleated erythroid cell of claim 1 , wherein the PAH is intracellular.
6. The enucleated erythroid cell of claim 1 , wherein the PAH is on the surface of the enucleated erythroid cell.
7. The enucleated erythroid cell of claim 1 , which is a reticulocyte.
8. The enucleated erythroid cell of claim 1 , which is a human enucleated erythroid cell.
9. The enucleated erythroid cell of claim 1 , which exhibits an increase in PAH activity of at least 2-fold relative to that of an enucleated erythroid cell that does not comprise the exogenous polypeptide.
10. The enucleated erythroid cell of claim 1 , wherein the PAH converts phenylalanine to tyrosine.
11. A pharmaceutical composition comprising a plurality of the enucleated erythroid cells of claim 1 and a pharmaceutically acceptable carrier.
12. The pharmaceutical composition of claim 11 , which comprises a population of erythroid cells that is greater than 60% enucleated.
13. The pharmaceutical composition of claim 11 , which comprises from 1×10 10 to 1×10 14 enucleated erythroid cells.
14. A method of treating phenylketonuria, the method comprising administering intravenously to a subject in need thereof the pharmaceutical composition of claim 11 , thereby treating said phenylketonuria.
15. A method of reducing phenylalanine concentration in the blood of a subject, the method comprising administering intravenously to a subject in need thereof the pharmaceutical composition of claim 11 , thereby reducing phenylalanine concentration in the blood of the subject.
16. The enucleated erythroid cell of claim 1 , which is a mature erythrocyte.
17. The method of claim 14 , wherein the concentration of phenylalanine in the blood is decreased by at least about 5% during part or the entirety of a treatment period.
18. The method of claim 14 , wherein the subject is deficient in phenylalanine ammonia hydroxylase (PAH) activity.
19. The method of claim 18 , wherein the deficiency in PAH activity is associated with a PAH enzyme mutation.
20. The method of claim 15 , wherein the subject has benign hyperphenylalaninemia or phenylketonuria.
21. The pharmaceutical composition of claim 11 , wherein at least 20% of enucleated erythroid cells in the pharmaceutical composition comprise the exogenous polypeptide.
22. The enucleated erythroid cell of claim 1 , wherein the exogenous nucleic acid comprises DNA.
23. The enucleated erythroid cell of claim 1 , wherein the exogenous nucleic acid comprises RNA.
24. The enucleated erythroid cell of claim 1 , wherein the exogenous polypeptide consists essentially of PAH.
25. The enucleated erythroid cell of claim 1 , wherein the exogenous polypeptide consists of PAH.
26. The enucleated erythroid cell of claim 1 , wherein the exogenous polypeptide is not fused to an endogenous polypeptide.
27. The method of claim 14 , wherein the pharmaceutical composition is administered intravenously to the subject.