Aav-mediated delivery of atp1a3 genes to central nervous system

Abstract

This invention relates to a transgenic vector for transducing cells of a mammal's CNS and transgenically expressing a protein in the mammal's CNS. The transgenic vector comprises a virus-derived vector, a nucleic acid sequence encoding the protein, and an endogenous ATP1A3 promoter sequence. This invention also relates to a composition comprising a recombinant AAV vector comprising a nucleic acid sequence encoding a ATP1A3 protein, in a form compatible with administration into the CNS. The invention also relates to a method for treating a subject having a neurological disorder associated with mutations in the ATP1A3 gene and a method for delivering a transgenic ATP1A3 DNA to the central nervous system of a mammal by administering the recombinant AAV vector into the mammal's CNS.

We claim: 
     
         1 . A method for treating a subject having alternating hemiplegia of childhood (AHC), the method comprising:
 administering into the central nervous system (CNS) of the subject a recombinant adeno-associated virus (AAV) vector composition comprising a nucleic acid sequence encoding a ATP1A3 protein, and a promoter sequence that renders the expression of the ATP1A3 protein-coding nucleic acid sequence specific to the central nervous system; wherein the AAV vector is AAV9, and wherein the promoter is a human neuron-specific promoter with a strong bias towards inhibitory neuron (P hSyn ),   thereby restoring some degree of neurological function in said subject.   
     
     
         2 . The method of  claim 1 , wherein the recombinant AAV vector is administered to one or more CNS components selected from the group consisting of cerebrospinal fluid (CSF), cisterna magna (cerebellomedullaris cistern), cerebral ventricles, the spinal cord, brainstem, hippocampus, cerebellum, cerebral cortex, diencephalon, and telencephalon. 
     
     
         3 . The method of  claim 1 , wherein the administration is via intracisternal, intrathecal, or intracerebroventricular injection. 
     
     
         4 . The method of  claim 1 , wherein the subject is human. 
     
     
         5 . The method of  claim 1 , wherein the recombinant AAV vector composition further comprising one or more nucleic acid regulatory sequences, linked directly or indirectly to the ATP1A3 protein-coding nucleic acid sequence. 
     
     
         6 . The method of  claim 5 , wherein the nucleic acid regulatory sequence comprises a sequence to regulate ribosome binding and/or translation efficiency of the ATP1A3 gene. 
     
     
         7 . The method of  claim 6 , wherein the sequence to regulate ribosome binding and/or translation efficiency is a Kozak sequence. 
     
     
         8 . The method of  claim 5 , wherein the nucleic acid regulatory sequence comprises a 3′-UTR sequence that contains a polyadenylation signal sequence. 
     
     
         9 . The method of  claim 8 , wherein the 3′-UTR sequence is a rabbit beta globin polyadenylation signal sequence (rBGpA). 
     
     
         10 . The method of  claim 5 , wherein the recombinant AAV vector comprises AAV9/P hSyn -ATP1A3 cDNA-rBGpA. 
     
     
         11 . The method of  claim 1 , wherein the AAV9 vector has a single-stranded DNA genome. 
     
     
         12 . The method of  claim 1 , wherein the recombinant AAV vector comprises SEQ ID NO: 3. 
     
     
         13 . The method of  claim 1 , wherein the recombinant AAV vector comprises SEQ ID NO: 4. 
     
     
         14 . The method of  claim 1 , wherein the recombinant AAV vector comprises SEQ ID NO: 5. 
     
     
         15 . The method of  claim 1 , wherein the recombinant AAV vector comprises SEQ ID NO: 6.