Gene therapy vectors for adrenoleukodystrophy and adrenomyeloneuropathy

1. A composition comprising one or more cells that comprise a vector selected from the group consisting of:

a) a vector comprising from 5′ to 3′:

(i) a left (5′) retroviral LTR;

(ii) a central polypurine tract/DNA flap (cPPT/FLAP);

(iii) a retroviral export element;

(iv) a promoter active in a microglial cell, operably linked to a polynucleotide encoding an ATP-binding cassette, sub-family D, member 1 (ABCD1) polypeptide; and

(v) a right (3′) retroviral LTR;

wherein the vector does not comprise a post-transcriptional regulatory element;

b) a lentiviral vector comprising from 5′ to 3′:

(i) a left (5′) LTR;

(ii) a cPPT/FLAP;

(iii) an RRE;

(iv) a MND promoter operably linked to a polynucleotide encoding a human ABCD1 poly peptide;

(v) a right (3′) LTR; and

(vi) a polyadenylation sequence;

wherein the sector does not comprise a post-transcriptional regulatory element;

c) a lentiviral vector comprising from 5′ to 3′:

(i) a left (5′) HIV-1 LTR;

(ii) a Psi (Ψ) packaging signal;

(iii) a cPPT/FLAP;

(iv) an RRE;

(v) a MND promoter, operably linked to a cDNA encoding a human ABCD1 polypeptide;

(vi) a right (3′) self-inactivating (SIN) HIV-1 LTR; and

(vii) a rabbit β-globin polyadenylation sequence;

wherein the vector does not comprise a post-transcriptional regulatory element;

d) a vector comprising from 5′ to 3′:

(i) a left (5′) retroviral LTR;

(ii) a central polypurine tract/DNA flap (cPPT/FLAP);

(iii) a retroviral export element;

(iv) a promoter active in a microglial cell, operably linked to a polynucleotide encoding an ATP-binding cassette, sub-family D, member 1 (ABCD1) polypeptide;

(v) a right (3′) retroviral LTR;

(vi) a polyadenylation (polyA) sequence; and

wherein the vector does not comprise a post-transcriptional regulatory element; and

e) a lentiviral vector comprising from 5′ to 3′:

(i) a left (5′) HIV-1 LTR;

(ii) a Psi (Ψ) packaging signal;

(iii) a cPPT/FLAP;

(iv) an RRE;

(v) a MND promoter, operably linked to a cDNA encoding a human ABCD1 polypeptide;

(vi) a right (3′) self-inactivating (SIN) HIV-1 LTR; and

(vii) a polyadenylation (polyA) sequence;

wherein the vector does not comprise a post-transcriptional regulatory element.

2. The composition of claim 1 , wherein the composition comprises an aqueous solution.

3. The composition of claim 1 , wherein the composition comprises an isotonic aqueous solution suitable for parental administration to a human.

4. The composition of claim 1 , wherein the composition comprises a cell culture medium.

5. The composition of claim 1 , wherein the one or more cells are embryonic stem cells, somatic stem cells, or progenitor cells.

6. The composition of claim 5 , wherein the somatic stem cells are bone marrow stem cells, umbilical cord stem cells, or mesenchymal stem cells.

7. The composition of claim 5 , wherein the somatic stem cells are hematopoietic stem cells.

8. The composition of claim 1 , wherein, in the vector:

a) the promoter of: the left (5′) LTR, the left (5′) retroviral LTR, or left (5′) HIV-1 LTR, is replaced with a heterologous promoter selected from the group consisting of: a cytomegalovirus (CMV) promoter, a Rous Sarcoma Virus (RSV) promoter, or a Simian Virus 40 (SV40) promoter;

b) the right (3′) LTR or the right (3′) retroviral LTR is a self-inactivating (SIN) LTR;

c) the promoter active in a microglial cell comprises a myeloproliferative sarcoma virus enhancer, negative control region deleted, d1587rev primer-binding site substituted (MND) promoter or transcriptionally active fragment thereof;

d) the polynucleotide encoding the ABCD1 polypeptide is a cDNA;

e) the polynucleotide encoding the ABCD1 polypeptide is a cDNA that comprises an optimized Kozak sequence; or

f) the polyadenylation sequence is a bovine growth hormone polyadenylation signal or a rabbit β-globin polyadenylation sequence.

9. A pharmaceutical composition comprising a pharmaceutically acceptable excipient, carrier, or diluent and one or more cells that comprise a vector selected from the group consisting of:

a) a vector comprising from 5′ to 3′:

(i) a left (5′) retroviral LTR;

(ii) a central polypurine tract/DNA flap (cPPT/FLAP);

(iii) a retroviral export element;

(iv) a promoter active in a microglial cell, operably linked to a polynucleotide encoding an ATP-binding cassette, sub-family D, member 1 (ABCD1) polypeptide; and

(v) a right (3′) retroviral LTR;

wherein the vector does not comprise a post-transcriptional regulatory element;

b) a lentiviral vector comprising from 5′ to 3′:

(i) a left (5′) LTR;

(ii) a cPPT/FLAP;

(iii) an RRE;

(iv) a MND promoter operably linked to a polynucleotide encoding a human ABCD1 polypeptide;

(v) a right (3′) LTR; and

(vi) a polyadenylation sequence;

wherein the vector does not comprise a post-transcriptional regulatory element;

c) a lentiviral vector comprising from 5′ to 3′:

(i) a left (5′) HIV-1 LTR;

(ii) a Psi (Ψ) packaging signal;

(iii) a cPPT/FLAP;

(iv) an RRE;

(v) a MND promoter, operably linked to a cDNA encoding a human ABCD1 polypeptide;

(vi) a right (3′) self-inactivating (SIN) HIV-1 LTR; and

(vii) at rabbit β-globin polyadenylation sequence;

wherein the vector does not comprise a post-transcriptional regulatory element;

d) a vector comprising from 5′ to 3′:

(i) a left (5′) retroviral LTR;

(ii) a central polypurine tract/DNA flap (cPPT/FLAP);

(iii) a retroviral export element;

(iv) a promoter active in a microglial cell, operably linked to a polynucleotide encoding an ATP-binding cassette, sub-family D, member 1 (ABCD1) polypeptide;

(v) a right (3′) retroviral LTR;

(vi) a polyadenylation (polyA) sequence; and

wherein the vector does not comprise a post-transcriptional regulatory element, and

e) a lentiviral vector comprising from 5′ to 3′:

(i) a left (5′) HIV-1 LTR;

(ii) a Psi (•) packaging signal;

(iii) a cPPT/FLAP;

(iv) an RRE;

(v) a MND promoter, operably linked to a cDNA encoding a human ABCD1 polypeptide;

vi) a right (3′) self-inactivating (SIN) HIV-1 LTR; and

(vii) a polyadenylation (polyA) sequence;

wherein the vector does not comprise a post-transcriptional regulatory element.

10. The pharmaceutical composition of claim 9 , wherein the pharmaceutically acceptable excipient, carrier, or diluent comprises a sterile aqueous solution.

11. The pharmaceutical composition of claim 9 , wherein the pharmaceutically acceptable excipient, carrier, or diluent comprises a pharmaceutically acceptable cell culture medium.

12. The pharmaceutical composition of claim 9 , wherein the one or more cells are embryonic stem cells, somatic stem cells, or progenitor cells.

13. The pharmaceutical composition of claim 12 , wherein the somatic stem cells are bone marrow stem cells, umbilical cord stem cells, or mesenchymal stem cells.

14. The pharmaceutical composition of claim 12 , wherein the somatic stem cells are hematopoietic stem cells.

15. The pharmaceutical composition of claim 9 , wherein, in the vector:

a) the promoter of: the left (5′) LTR, the left (5′) retroviral LTR, or left (5′) HIV-1 LTR, is replaced with a heterologous promoter selected from the group consisting of: a cytomegalovirus (CMV) promoter, a Rous Sarcoma Virus (RSV) promoter, or a Simian Virus 40 (SV40) promoter;

b) the right (3′) LTR or the right (3′) retroviral LTR is a self-inactivating (SIN) LTR;

c) the promoter active in i microglial cell comprises a myeloproliferative sarcoma virus enhancer, negative control region deleted, d1587rev primer-binding site substituted (MND) promoter or transcriptionally active fragment thereof;

d) the polynucleotide encoding the ABCD1 polypeptide is a cDNA;

e) the polynucleotide encoding the ABCD1 polypeptide is a cDNA that comprises an optimized Kozak sequence; or

f) the polyadenylation sequence is a bovine growth hormone polyadenylation signal or a rabbit β-globin polyadenylation sequence.

16. A formulation comprising a pharmaceutically-acceptable solution and one or more cells comprising a vector selected from the group consisting of:

a) a vector comprising from 5′ to 3′:

(i) a left (5′) retroviral LTR;

(ii) a Psi (Ψ) packaging signal;

(iii) a central polypurine tract/DNA flap (cPPT/FLAP);

(iv) a retroviral export element;

(v) a promoter active in a microglial cell, operably linked to a polynucleotide encoding an ATP-binding cassette, sub-family D, member 1 (ABCD1) polypeptide; and

(vi) a right (3′) retroviral LTR;

wherein the vector does not comprise a post-transcriptional regulatory element;

b) a lentiviral vector comprising from 5′ to 3′:

(i) a left (5′) LTR;

(ii) a Psi (Ψ) packaging signal;

(iii) a cPPT/FLAP;

(iv) an RRE;

(v) a MND promoter operably linked to a poly nucleotide encoding a human ABCD1 polypeptide;

(vi) a right (3′) LTR; and

(vii) a polyadenylation sequence;

wherein the vector does not comprise a post-transcriptional regulatory element;

c) a lentiviral vector comprising from 5′ to 3′:

(i) a left (5′) HIV-1 LTR;

(ii) a Psi (Ψ) packaging signal;

(iii) a cPPT/FLAP;

(iv) an RRE;

(v) a MND promoter, operably linked to a cDNA encoding; a human ABCD1 polypeptide;

(vi) a right (3′) self-inactivating (SIN) HIV-1 LTR; and

(vii) a rabbit β-globin polyadenylation sequence;

wherein the sector does not comprise a post-transcriptional regulatory element;

d) a vector comprising from 5′ to 3′:

(i) a left (5′) retroviral LTR;

(ii) a cPPT/FLAP;

(iii) a retroviral export element;

(iv) a promoter active in a microglial cell, operably linked to a polynucleotide encoding an ATP-binding cassette, sub-family D, member 1 (ABCD1) polypeptide;

(v) a right (3′) retroviral LTR;

(vii) a polyadenylation (polyA) sequence; and

wherein the vector does not comprise a post-transcriptional regulatory element; and

e) a lentiviral vector comprising from 5′ to 3′:

(i) a left (5′) HIV-1 LTR;

(ii) a Psi (Ψ) packaging signal;

(iii) a cPPT/FLAP;

(iv) an RRE;

(v) a MND promoter operably linked to a cDNA encoding a human ABCD1 polypeptide;

(vi) a right (3′) self-inactivating (SIN) HIV-1 LTR; and

(vi) a polyadenylation (polyA) sequence;

wherein the vector does not comprise a post-transcriptional regulatory element.

17. The formulation of claim 16 , wherein the formulation comprises a sterile isotonic aqueous solution.

18. The formulation of claim 16 , wherein the one or more cells are embryonic stem cells, somatic stem cells, or progenitor cells.

19. The formulation of claim 18 , wherein the somatic stem cells are bone marrow stem cells, umbilical cord stem cells, or mesenchymal stem cells.

20. The formulation of claim 18 , wherein the somatic stem cells are hematopoietic stem cells.

21. The formulation of claim 16 , wherein, in the vector:

a) the promoter of the left (5′) LTR, the left (5′) retroviral LTR, or left (5′) HIV-1 LTR, is replaced with a heterologous promoter selected from the group consisting of: a cytomegalovirus (CMV) promoter, a Rous Sarcoma Virus (RSV) promoter, or a Simian Virus 40 (SV40) promoter;

b) the right (3′) LTR or the right (3′) retroviral LTR is a self-inactivating (SIN) LTR;

c) the promoter active in a microglial cell comprises a myeloproliferative sarcoma virus enhancer, negative control region deleted, d1587rev primer-binding site substituted (MND) promoter or transcriptionally active fragment thereof;

d) the polynucleotide encoding the ABCD1 polypeptide is a cDNA;

e) the polynucleotide encoding the ABCD1 polypeptide is a cDNA that comprises an optimized Kozak sequence; or

f) the polyadenylation sequence is a bovine growth hormone polyadenylation signal or a rabbit β-globin polyadenylation sequence.