Method for improving retroviral transduction and gene editing in hematopoietic stem cells using cyclosporine h (CsH)
Use of cyclosporin H (CsH) or a derivative thereof for increasing the efficiency of transduction of an isolated population of cells by a viral vector and/or increasing the efficiency of gene editing of an isolated population of cells when transduced by a viral vector.
1. A method of transducing a population of cells comprising the steps of:
(a) contacting the population of cells with cyclosporin H (CsH); and
(b) transducing the population of cells with a viral vector, wherein the viral vector is a lentiviral or gamma-retroviral vector and wherein the viral vector is pseudotyped to enter cells via an endocytosis-dependent mechanism,
wherein the population of cells comprises: (i) CD34+ HSPCs; (ii) CD14+ monocytes; (iii) CD4+ or CD3+ T cells; (iv) CD34+ bone marrow cells; or (v) CD34+ peripheral blood cells.
2. The method of claim 1 , wherein steps (a) and (b) are carried out in vitro or ex vivo.
3. The method of claim 1 , wherein the percentage of cells transduced by the vector is increased and/or the vector copy number per cell is increased.
4. The method of claim 1 , wherein the CsH is at a concentration of about 1-50 μM.
5. The method of claim 1 , wherein the population of cells is contacted with CsH in combination with rapamycin.
6. The method of claim 1 , wherein the population of cells is contacted with CsH in combination with prostaglandin E2.
7. The method of claim 1 , comprising a further step of enriching the population for CD34+/CD38− cells.
8. The method of claim 1 , wherein efficiency of transduction of the population of cells by the viral vector is increased and/or efficiency of gene editing of the population of cells when transduced by the viral vector is increased.
9. The method of claim 1 , wherein the method further comprises a wash step before or after transducing the population of cells with the viral vector, wherein the wash step substantially removes the CsH.
10. The method of claim 1 , wherein the T cells are CD4 + and/or CD3 + T cells.
11. The method of claim 1 , wherein the viral vector is a lentiviral vector.
12. The method of claim 6 , wherein the prostaglandin E2 is 16-16 dimethyl prostaglandin E2.
13. The method of claim 1 , wherein the viral vector is an integration-defective lentiviral vector (IDLV).
14. The method of claim 1 , wherein the cells are stimulated cells.
15. The method of claim 1 , wherein the population of cells is contacted with CsH in combination with another agent capable of increasing the efficiency of transduction.
16. The method of claim 1 , wherein the viral vector comprises a nucleotide of interest, and wherein
the nucleotide of interest encodes a chimeric antigen receptor (CAR).
17. The method of claim 1 , wherein the viral vector is a VSV-g pseudotyped vector.