Increased transduction using ABC transporter substrates and/or inhibitors
The present invention relates to improvements in the ability to transduce a retroviral vector borne nucleic acid into cells expressing ABC transporters by use of a substrate and/or inhibitor of said transporter. Compositions and kits relating to the practice of the methods are also disclosed. Methods to determine the level of increased transduction provided by a substrate and/or inhibitor compound are also provided.
1 . A method for increasing the transduction of an ABC transporter expressing cell with a retroviral vector, said method comprising
a) contacting said cell with a compound that is a substrate of said transporter; and
b) contacting said cell with said vector.
2 . The method of claim 1 , wherein said retroviral vector is a lantiviral vector.
3 . The method of claim 1 , wherein said compound is both a substrate and an inhibitor of said transporter.
4 . The method of claim 2 , wherein said compound is both a substrate and an inhibitor of said transporter.
5 . The method of any one of claims 1 - 4 wherein said cell is CD34+.
6 . The method of any one of claims 1 - 4 wherein said cells are hematopoietic stem cells.
7 . The method of any one of claims 1 - 4 wherein said cells are primary cells.
8 . The method of claim 7 wherein said cells are from bone marrow, cord blood, or peripheral blood.
9 . The method of any one of claims 1 - 4 wherein said compound is selected from verapamil, diltiazem, quinindine, and ritonavir.
10 . The method of claim 2 or 4 wherein said lentiviral vector is derived from HIV or SIV.
11 . The method of any one of claims 1 - 4 wherein said vector is pseudotyped with VSV-G:
12 . The method of claim 1 or 3 wherein said vector is derived from an oncoretrovirus.
13 . The method of claim 1 or 3 wherein said vector is derived from a Spumavirus.
14 . A method of determining the level of increased transduction of a retroviral vector in a cell mediated by an ABC transporter substrate or inhibitor compound, said method comprising
a) providing a population of said cell;
b) contacting one or more cells of said population with said vector in the presence of said compound to produce a first contacted cell or cells and contacting a second cell or cells of said population with said vector in the absence of said compound to produce a second contacted cell or cells; and
c) determining the level of transduction with said vector in said first contacted cell or cells and in said second contacted cell or cells.
15 . The method of claim 14 wherein said population of cells are primary cells.
16 . The method of claim 15 wherein said cells are from a human being.
17 . The method of any one of claims 14 - 16 wherein said cell is known to express an ABC transporter inhibitor.
18 . The method of claim 14 wherein said vector is a lentiviral vector.
19 . The method of claim 15 wherein said vector is a lentiviral vector.
20 . The method of claim 16 wherein said vector is a lentiviral vector.
21 . The method of claim 17 wherein said vector is a lentiviral vector.
22 . Use of a retroviral vector for the preparation of a medicament for increasing the transduction of an ABC tranporter expressing cell.