Adeno-Associated Virus Virions for Treatment of Epilepsy

Abstract

Provided is a novel gene therapy means for neurological diseases including epilepsy. The present invention provides: a recombinant adeno-associated virus vector for use in the treatment of neurological diseases including epilepsy, which comprises a polynucleotide encoding a protein capable of improving the excitation-inhibiting function of an inhibitory synapse in vivo, preferably neuroligin-2 protein; a pharmaceutical composition comprising said recombinant vector; and others. The present invention also provides a method for treating a disease such as epilepsy using the recombinant vector.

1 . A recombinant adeno-associated virus vector, wherein the virus vector comprises a polynucleotide encoding a protein for improving an excitation-inhibiting function of inhibitory synapses in a living subject, and is used for treatment of a disease selected from the group consisting of epilepsy, schizophrenia, autism spectrum disorder, mental retardation, anxiety, manic-depressive psychosis, migraine, phobic and compulsive symptoms, drug addiction, Angelman syndrome, dyskinesia, dystonia, Alzheimer's disease, and developmental disorders (attention deficit hyperactivity disorder and Asperger's syndrome),
 wherein the polynucleotide comprises a nucleotide sequence encoding a neuroligin 2 protein which comprises the amino acid sequence of SEQ ID NO: 2, 4 or 6, or an amino acid sequence having about 90% or more identity with said amino acid sequence and capable of binding to neurexin, and   wherein the recombinant adeno-associated virus vector comprises:   a protein having a variant amino acid sequence in which tyrosine at position 445 in the amino acid sequence of a wild-type AAV1 capsid protein is substituted with phenylalanine;   a protein having a variant amino acid sequence in which tyrosine at position 445 in the amino acid sequence of a wild-type AAV2 capsid protein is substituted with phenylalanine; or   a protein having a variant amino acid sequence in which tyrosine at position 446 in the amino acid sequence of a wild-type AAV9 capsid protein is substituted with phenylalanine.   
     
     
         2 . (canceled) 
     
     
         3 . The recombinant adeno-associated virus vector according to  claim 1 , wherein the disease is epilepsy. 
     
     
         4 . (canceled) 
     
     
         5 . The recombinant adeno-associated virus vector according to  claim 1 , wherein the polynucleotide comprises a promoter sequence selected from the group consisting of a synapsin I promoter sequence, a myelin basic protein promoter sequence, a neuron specific enolase promoter sequence, a calcium/calmodulin-dependent protein kinase II (CMKII) promoter sequence, a tubulin α1 promoter sequence, a platelet-derived growth factor β chain promoter sequence, a glial fibrillary acidic protein (GFAP) promoter sequence, a L7 promoter (cerebellar Purkinje cell specific promoter) sequence, a glial fibrillary acidic protein (hGfa2) promoter sequence, and a glutamate receptor delta 2 promoter (cerebellar Purkinje cell specific promoter) sequence, and a glutamic acid decarboxylase (GAD65/GAD67) promoter sequence. 
     
     
         6 . The recombinant adeno-associated virus vector according to  claim 1 , wherein the polynucleotide comprises an inverted terminal repeat (ITR) selected from the group consisting of AAV1, AAV2, AAV3, AAV4, AAV8, and AAV9. 
     
     
         7 . The recombinant adeno-associated virus vector according to  claim 1 , wherein the polynucleotide further comprises a polynucleotide for inhibiting the excitation of excitatory synapses. 
     
     
         8 . A pharmaceutical composition, comprising the recombinant adeno-associated virus recombinant vector according to  claim 1 . 
     
     
         9 . The pharmaceutical composition according to  claim 8 , which is administered intracerebrally. 
     
     
         10 . The pharmaceutical composition according to  claim 8 , which is administered intrathecally. 
     
     
         11 . The pharmaceutical composition according to  claim 8 , which is administered peripherally. 
     
     
         12 . The pharmaceutical composition according to  claim 8 , which is used in combination with a chemotherapeutic agent for a neuropsychiatric disease.