Method for treating adenosine deaminase severe combined imunodeficiency
The present invention relates to the prevention and/or treatment of ADA-SCID, in a patient.
1. A method of treating adenosine deaminase severe combined immunodeficiency (ADA-SCID) in a human patient in need thereof, comprising administering a therapeutically effective amount of a composition comprising a host human hematopoietic stem cell (HSC) or a cell population of human HSCs to the blood and/or bone marrow of said patient, wherein said host cell or cell population contains a vector or expression cassette comprising a transgene encoding a human adenosine deaminase (hADA) operably linked to a regulator region comprising an elongation factor 1-alpha short isoform (EFS) promoter; wherein said transgene is expressed in peripheral blood mononuclear cells (PBMCs) of said patient and wherein the transgene expression is elevated for at least 6 months after administration of the composition and is elevated relative to expression of a transgene encoding said hADA operably linked to a MND promoter when normalized per vector copy number.
2. The method of claim 1 , wherein:
(a) the vector or expression cassette is present at a copy number of 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 copies per single cell; or
(b) the vector or expression cassette is present at a median copy number of from 0.5 to 6 in said cell population.
3. The method of claim 1 , wherein said vector is a self-inactivating lentiviral vector comprising said transgene operably linked to said EFS promoter, wherein said vector further comprises a woodchuck hepatitis post-transcriptional regulatory element (wPRE) element.
4. The method of claim 1 , wherein the transgenes are codon-optimized for expression in a human cell.
5. The method of claim 1 , wherein the vector is a self-inactivating human immunodeficiency virus 1 (HIV-1) lentiviral vector.
6. The method of claim 1 , wherein the transgene encoding the hADA operably linked to the regulatory region comprising the EFS promoter has a sequence of SEQ ID NO: 1 or a variant thereof having at least 95% sequence identity to SEQ ID NO: 1.
7. The method of claim 1 , wherein the cell population further comprises peripheral blood stem cells (PBSCs).
8. The method of claim 1 , wherein said host cell or cell population is isolated prior to pre-stimulation in medium supplemented with Flt3L, IL-3, TPO and SCF.
9. The method of claim 1 , wherein expression of hADA and/or metabolic activity of hADA is corrected to levels present in a human that does not have ADA-SCID or levels above those found in a human that does not have ADA-SCID.
10. The method of claim 3 , wherein expression of hADA and/or activity of hADA is higher than expression of hADA and/or activity of hADA in a patient with ADA-SCID that received the transgene encoding hADA operably linked to the regulatory region comprising the EFS promoter delivered by a gamma retroviral vector present at the same average copy number as the lentiviral vector.
11. The method of claim 9 , wherein the expression of hADA and/or activity of hADA is corrected in the PMBC and/or red blood cells.
12. The method of claim 1 , wherein at least one immunological defect associated with ADA-SCID is corrected in said patient.
13. The method of claim 12 , wherein the CD3 count, CD4 count, CD8 count, naïve T cell count, T-cell receptor excision circles (TREC) levels and/or immunoglobulin levels are corrected.
14. The method of claim 1 , wherein expression of the transgene encoding the hADA operably linked to the EFS promoter is sustained for at least 12, 24, or 48 months post-administration of the composition.
15. The method of claim 1 , wherein the host cell or cell population is derived from the same patient, an individual who is related to the patient, or an individual who is a tissue type match for the patient.
16. The method of claim 1 , wherein the host cell or cell population is derived from an individual with a different genetic background from the patient to which it is administered.
17. The method of claim 1 , wherein: (a) the vector or expression cassette comprises a lentivirus genome; and (b) the transgene encoding hADA operably linked to the EFS promoter is flanked by a long terminal repeat (LTR) and a central polypurine tract (cPPT) at the 5′ end and a wPRE sequence and a LTR lacking a U3 region at the 3′ end.
18. The method of claim 1 , wherein the composition further comprises a pharmaceutically acceptable excipient.
19. The method of claim 5 , where the host cell or cell population is made by a method comprising introducing into said host cell or cell population a self-inactivating HIV-1 vector, wherein said vector encodes hADA under the control of an EFS promoter, and wherein the vector comprises a wPRE element.
20. The method of claim 19 , wherein the HIV-1 vector comprises the sequence of SEQ ID NO: 2 or a variant thereof having at least 85% identity to SEQ ID NO: 2.
21. The method of claim 19 , comprising isolating the cell or cell population from a human.
22. The method of claim 21 , further comprising culturing the isolated cell or cell population.
23. The method of claim 22 , wherein the isolated cell or cell population is derived from the same patient, an individual who is related to the patient, an individual who is a tissue type match for the patient, or an individual with a different genetic background from the patient to which the composition is administered.