Gene Therapy For Neurodegenerative Disorders

Abstract

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 - 19 . (canceled) 
     
     
         20 . A recombinant virus comprising a self-complementary adeno-associated virus (scAAV) vector comprising:
 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 polynucleotide encoding the SMN protein comprising the amino acid sequence of SEQ ID NO: 2 is expressed in motor neurons when administered to a subject having spinal muscular atrophy (SMA).   
     
     
         21 . The recombinant virus of  claim 20 , wherein the second AAV2 ITR comprises a mutated terminal resolution site. 
     
     
         22 . The recombinant virus of  claim 21 , wherein the mutated terminal resolution site comprises a deletion of nucleotides in the terminal resolution site sequence. 
     
     
         23 . The recombinant virus of  claim 20 , wherein the first and second ITRs are wild-type, wherein the scAAV vector further comprises a third AAV2 inverted terminal repeat (ITR) with a mutated terminal resolution site comprising a deletion of nucleotides in the terminal resolution site sequence, and further wherein the first and second AAV2 ITRs are located at each end of the scAAV vector and the third AAV2 ITR is located in between the first and the second ITR. 
     
     
         24 . The recombinant virus of  claim 20 , further comprising a bovine growth hormone (BGH) polyadenylation site. 
     
     
         25 . A composition comprising a recombinant adeno-associated virus (rAAV) virion and a pharmaceutical excipient, wherein the rAAV virion comprises a self-complementary adeno-associated virus (scAAV) vector comprising:
 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 virion comprises an AAV8 or AAV9 capsid. 
     
     
         26 . The composition of  claim 25 , wherein the polynucleotide encoding a SMN protein comprising the amino acid sequence of SEQ ID NO: 2 is expressed in motor neurons when the composition is administered to subject having spinal muscular atrophy (SMA). 
     
     
         27 . The composition of  claim 25 , wherein the composition comprises a therapeutically effective amount of rAAV virions. 
     
     
         28 . The composition of  claim 25 , wherein the composition comprises about 10 8  to 10 14  gp/mL of rAAV virions. 
     
     
         29 . The composition of  claim 25 , wherein the second AAV2 ITR comprises a mutated terminal resolution site. 
     
     
         30 . The composition of  claim 26 , wherein the mutated terminal resolution site comprises a deletion of nucleotides in the terminal resolution site sequence. 
     
     
         31 . The composition of  claim 25 , wherein the first and second ITRs are wild-type, wherein the scAAV vector further comprises a third AAV2 inverted terminal repeat (ITR) with a mutated terminal resolution site comprising a deletion of nucleotides in the terminal resolution site sequence, and further wherein the first and second AAV2 ITRs are located at each end of the scAAV vector and the third AAV2 ITR is located between the first and the second ITR. 
     
     
         32 . The composition of  claim 25 , wherein the scAAV vector further comprises a bovine growth hormone (BGH) polyadenylation site. 
     
     
         33 . A kit comprising:
 a) a container comprising a composition comprising a recombinant adeno-associated virus (rAAV) virion and a pharmaceutical excipient, the rAAV virion comprising a self-complementary adeno-associated virus (scAAV) vector comprising:
 i) a first AAV2 inverted terminal repeat (ITR); 
 ii) a cytomegalovirus enhancer/chicken-β actin (CBA) promoter; 
 iii) a polynucleotide encoding a survival motor neuron (SMN) protein comprising the amino acid sequence of SEQ ID NO: 2; and 
 iv) a second AAV2 ITR, 
 wherein the rAAV virion comprises an AAV8 or AAV9 capsid, and 
   b) instructions for administering the composition to a subject with spinal muscular atrophy (SMA).   
     
     
         34 . The kit of  claim 33 , wherein the polynucleotide encoding a SMN protein comprising the amino acid sequence of SEQ ID NO: 2 is expressed in motor neurons when the composition is administered to subject having spinal muscular atrophy (SMA). 
     
     
         35 . The kit of  claim 33 , wherein the composition comprises a therapeutically effective amount of rAAV virions. 
     
     
         36 . The kit of  claim 33 , wherein the composition comprises about 10 8  to 10 14  gp/mL of rAAV virions. 
     
     
         37 . The kit of  claim 33 , wherein the second AAV2 ITR comprises a mutated terminal resolution site. 
     
     
         38 . The kit of  claim 37 , wherein the mutated terminal resolution site comprises a deletion of nucleotides in the terminal resolution site sequence. 
     
     
         39 . The kit of  claim 33 , wherein the first and second ITRs are wild-type, wherein the scAAV vector further comprises a third AAV2 inverted terminal repeat (ITR) with a mutated terminal resolution site comprising a deletion of nucleotides in the terminal resolution site sequence, and further wherein the first and second AAV2 ITRs are located at each end of the scAAV vector and the third AAV2 ITR is located between the first and the second ITR. 
     
     
         40 . The kit of  claim 33 , wherein the scAAV vector further comprises a bovine growth hormone (BGH) polyadenylation site.