Gene therapy for neurodegenerative disorders
Compositions and methods for treating disorders affecting motor function, such as motor function affected by disease or injury to the brain and/or spinal cord, are disclosed.
1. A method of treating spinal muscular atrophy (SMA) comprising administering to a subject in need thereof a therapeutically effective amount of recombinant adeno-associated virus (rAAV) virions comprising a self-complementary adeno-associated virus (scAAV) vector comprising a heterologous nucleic acid construct, wherein the heterologous nucleic acid construct comprises:
a. a first AAV2 inverted terminal repeat (ITR);
b. a cytomegalovirus enhancer/chicken-β actin (CBA) promoter;
c. a polynucleotide encoding a survival motor neuron (SMN) protein comprising the amino acid sequence of SEQ ID NO: 2; and
d. a second AAV2 ITR, and
wherein the rAAV virions comprise an AAV8 or AAV9 capsid, and
wherein the scAAV virions are administered via intrathecal injection.
2. The method of claim 1 , wherein the first or the second ITR is mutated.
3. The method of claim 2 , wherein the first or the second ITR comprises a mutation in a terminal resolution sequence.
4. The method of claim 1 , wherein a single-dose of the rAAV virions is administered.
5. A method of expressing survival motor neuron (SMN) protein in motor neurons of the spinal cord comprising administering to a subject in need thereof a therapeutically effective amount of recombinant adeno-associated virus (rAAV) virions, wherein the rAAV virions comprise a self-complementary adeno-associated virus (scAAV) vector comprising a heterologous nucleic acid construct,
wherein the subject has spinal muscular atrophy (SMA), wherein the heterologous nucleic acid construct comprises:
a. a first AAV2 inverted terminal repeat (ITR);
b. a cytomegalovirus enhancer/chicken-β actin (CBA) promoter;
c. a polynucleotide encoding a survival motor neuron (SMN) protein comprising the amino acid sequence of SEQ ID NO: 2; and
d. a second AAV2 ITR, and
wherein the rAAV virions comprise an AAV8 or AAV9 capsid, and
wherein the rAAV virions are administered via intrathecal injection.
6. The method of claim 5 , wherein the first or the second ITR is mutated.
7. The method of claim 6 , wherein the first or the second ITR comprises a mutation in a terminal resolution sequence.
8. The method of claim 5 , wherein a single-dose of the rAAV virions is administered.
9. The method of claim 5 , wherein the motor neurons of the spinal cord are in the dorsal and ventral horns of the spinal cord.
10. The method of claim 5 , wherein the motor neurons of the spinal cord are in the lumbar, thoracic and/or cervical segments of the spinal cord.
11. A method of increasing survival, comprising administering to a subject in need thereof a therapeutically effective amount of recombinant adeno-associated virus (rAAV) virions,
wherein the subject has spinal muscular atrophy (SMA),
wherein the rAAV virions comprise a self-complementary adeno-associated virus (scAAV) vector comprising a heterologous nucleic acid construct, wherein the heterologous nucleic acid construct comprises:
a. a first AAV2 inverted terminal repeat (ITR);
b. a cytomegalovirus enhancer/chicken-β actin (CBA) promoter;
c. a polynucleotide encoding a survival motor neuron (SMN) protein comprising the amino acid sequence of SEQ ID NO: 2; and
d. a second AAV2 ITR,
wherein the rAAV virions comprise an AAV8 or AAV9 capsid, and
wherein the rAAV virions are administered via intrathecal injection.
12. The method of claim 11 , wherein a single-dose of the rAAV virions is administered.
13. The method of claim 1 , wherein about 10 6 to about 10 15 rAAV virions are administered.
14. The method of claim 5 , wherein about 10 6 to about 10 15 rAAV virions are administered.
15. The method of claim 11 , wherein about 10 6 to about 10 15 rAAV virions are administered.
16. The method of claim 11 , wherein the first or the second ITR is mutated.
17. The method of claim 16 , wherein the first or the second ITR comprises a mutation in a terminal resolution sequence.
18. The method of claim 2 , wherein the mutation comprises a deletion at the terminal resolution site of the first or the second ITR.
19. The method of claim 6 , wherein the mutation comprises a deletion at the terminal resolution site of the first or the second ITR.
20. The method of claim 16 , wherein the mutation comprises a deletion at the terminal resolution site of the first or the second ITR.