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 . A non-hypotonically loaded enucleated erythroid cell comprising an exogenous polypeptide comprising cystathionine beta synthase (CBS) or a functional fragment thereof.
2 . The non-hypotonically loaded enucleated erythroid cell of claim 1 , further comprising a second exogenous polypeptide comprising a transporter for a substrate of the CBS.
3 . The non-hypotonically loaded enucleated erythroid cell of claim 1 , which comprises at least 1,000 copies of the exogenous polypeptide.
4 . The non-hypotonically loaded enucleated erythroid cell of claim 1 , which comprises at least 10,000 copies of the exogenous polypeptide.
5 . The non-hypotonically loaded enucleated erythroid cell of claim 1 , which comprises at least 50,000 copies of the exogenous polypeptide.
6 . The non-hypotonically loaded enucleated erythroid cell of claim 1 , which comprises at least 100,000 copies of the exogenous polypeptide.
7 . The non-hypotonically loaded enucleated erythroid cell of claim 1 , wherein the CBS is intracellular.
8 . The non-hypotonically loaded enucleated erythroid cell of claim 1 , wherein the CBS is on the surface of the enucleated erythroid cell.
9 . The non-hypotonically loaded enucleated erythroid cell of claim 1 , which is a reticulocyte or mature erythrocyte.
10 . The non-hypotonically loaded enucleated erythroid cell of claim 1 , which 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, or the precursor thereof, under conditions suitable for enucleation of the nucleated erythroid cell and for production of the exogenous polypeptide.
11 . The non-hypotonically loaded enucleated erythroid cell of claim 10 , wherein the process of producing further comprises introducing the exogenous nucleic acid into the nucleated erythroid cell, or the precursor thereof.
12 . The non-hypotonically loaded enucleated erythroid cell of claim 10 , wherein the exogenous nucleic acid is situated in a lentiviral vector.
13 . The non-hypotonically loaded enucleated erythroid cell of claim 1 , which exhibits an increase in CBS activity of at least 2-fold relative to that of an enucleated erythroid cell that does not comprise the exogenous polypeptide.
14 . The non-hypotonically loaded enucleated erythroid cell of claim 1 , wherein the substrate is homocysteine.
15 . A pharmaceutical composition comprising a plurality of the non-hypotonically loaded enucleated erythroid cells of claim 1 and a pharmaceutically acceptable carrier.
16 . The pharmaceutical composition of claim 15 , which comprises a population of erythroid cells that is greater than 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99%, 99.5% or 99.9% enucleated.
17 . A method of reducing homocysteine levels in a subject, comprising administering to the subject the pharmaceutical composition of claim 15 , thereby reducing homocysteine levels in the subject.
18 . A method of treating homocystinuria, the method comprising administering intravenously to a subject in need thereof the pharmaceutical composition of claim 15 , thereby treating said homocystinuria.