Gene therapy vectors for adrenoleukodystrophy and adrenomyeloneuropathy
The present invention provides compositions comprising retroviral vectors, transduced cells, and methods of using the same for gene therapy. In particular, the present invention relates to lentiviral vectors and cells transduced with those vectors to provide gene therapy to subjects having an adrenoleukodystrophy and/or adrenomyeloneuropathy.
1. A method of gene therapy comprising intravenously administering to a subject having adrenoleukodystrophy or adrenomyeloneuropathy, one or more CD34 + hematopoietic stem or progenitor 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 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) 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;
wherein the gene therapy treats adrenoleukodystrophy or adrenomyeloneuropathy in the subject.
2. A method of treating adrenoleukodystrophy or adrenomyeloneuropathy, comprising intravenously administering to a subject one or more CD34 + hematopoietic stem or progenitor cells transduced with 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) 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;
wherein the treatment reduces or ameliorates of symptoms of adrenoleukodystrophy or adrenomyeloneuropathy in the subject, or the delays progression of adrenoleukodystrophy or adrenomyeloneuropathy in the subject.
3. The method of claim 1 or claim 2 , wherein the one or more CD34 + hematopoietic stem or progenitor cells are CD34 + hematopoietic stem cells.
4. The method of claim 3 , wherein the CD34 + hematopoietic stem cells are isolated from bone marrow, umbilical cord blood, or peripheral blood.
5. The method of claim 1 or claim 2 , wherein the subject has undergone chemoablative or radioablative bone marrow therapy.
6. The method of claim 1 or claim 2 , wherein the subject is a young male.
7. The method of claim 6 , wherein the young male is between 3 to 15 years of age.
8. The method of claim 1 or claim 2 , wherein, in the vector, 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 an Simian Virus 40 (SV40) promoter.
9. The method of claim 1 or claim 2 , wherein, in the vector, the right (3′) LTR or the right (3′) retroviral LTR is a self-inactivating (SIN) LTR.
10. The method of claim 8 , wherein, in the vector, the right (3′) LTR or the right (3′) retroviral LTR is a self-inactivating (SIN) LTR.
11. The method of claim 1 or claim 2 , wherein, in the vector, the promoter active in a microglial cell comprises a myeloproliferative sarcoma virus enhancer, negative control region deleted, dl587rev primer-binding site substituted (MND) promoter or transcriptionally active fragment thereof.
12. The method of claim 1 or claim 2 , wherein, in the vector, the polynucleotide encoding the ABCD1 polypeptide is a cDNA.
13. The method of claim 1 or claim 2 , wherein, in the vector, the cDNA comprises an optimized Kozak sequence.
14. The method of claim 1 or claim 2 , wherein, in the vector, the polyadenylation sequence is a bovine growth hormone polyadenylation signal or a rabbit β-globin polyadenylation sequence.