Methods and compositions for the treatment of neurological disease

Abstract

The present invention relates to a method for delivering a nucleic acid sequence encoding neuropeptide Y, or a derivative or functional fragment thereof, to a mammalian nervous system target cell. The expression of exogenous NPY, or a derivative or a functional fragment thereof in the target cell(s) provides therapeutic benefit for subjects afflicted with a neurological disorder.

1 . A method for delivering a nucleic acid sequence to a mammalian nervous system target cell, wherein said nucleic acid sequence is expressible in the target cell for greater than three months, said method comprising administering an expression vector to the target cell, wherein said expression vector comprises a nucleic acid sequence encoding neuropeptide Y, or a derivative or functional fragment thereof. 
     
     
         2 . The method of  claim 1 , wherein the nucleic acid sequence encoding neuropeptide Y, or a derivative or functional fragment thereof, is expressed in said target cell either constitutively or under regulatable conditions. 
     
     
         3 . The method of  claim 2 , wherein expression of neuropeptide Y, or a derivative or functional fragment thereof, in said target cell alters neuronal excitability. 
     
     
         4 . The method of  claim 2 , wherein expression of neuropeptide Y, or a derivative or functional fragment thereof, in said target cell reduces neuronal excitability. 
     
     
         5 . The method of  claim 2 , wherein expression of neuropeptide Y, or a derivative or functional fragment thereof, in said target cell reduces symptoms associated with neuronal hyperexcitability. 
     
     
         6 . The method of  claim 1 , wherein the expression vector is a viral or a non-viral expression vector. 
     
     
         7 . The method of  claim 6 , wherein the viral expression vector is an adeno-associated virus (AAV) vector, a lentivirus vector, an adenovirus vector, or a herpes simplex virus (HSV) vector. 
     
     
         8 . The method of  claim 7 , wherein the viral expression vector is an AAV vector capable of transducing the target cell and said AAV vector is free of both wildtype and helper virus. 
     
     
         9 . The method of  claim 8 , wherein the AAV vector is a serotype 2 AAV vector or a chimeric serotype 1/2 AAV vector. 
     
     
         10 . The method of  claim 2 , wherein the nucleic acid sequence encoding neuropeptide Y, or a derivative or functional fragment thereof, is operably linked to an inducible regulatory sequence, wherein activation of said inducible regulatory sequence effects transcription of messenger RNA encoding neuropeptide Y from said nucleic acid sequence. 
     
     
         11 . The method of  claim 10 , wherein said inducible regulatory sequence renders NPY expression nervous system-specific or central nervous system-specific. 
     
     
         12 . The method of  claim 10 , wherein expression of neuropeptide Y is specific to a medial temporal lobe or temporal cortex of the central nervous system. 
     
     
         13 . The method of  claim 12 , wherein expression of neuropeptide Y in the medial temporal lobe is localized to the hippocampus and/or amygdala. 
     
     
         14 . The method of  claim 12 , wherein the expression is neural or glial specific. 
     
     
         15 . The method of  claim 1 , wherein the target cell is a mammalian cell of a mammalian order selected from the group consisting of Primata, Rodenta, Carnivora and Arteriodactyla. 
     
     
         16 . The method of  claim 15 , wherein the target cell is a human cell. 
     
     
         17 . The method of  claim 1 , wherein the target cell is in cell culture. 
     
     
         18 . The method of  claim 1 , wherein the target cell is in a living mammal. 
     
     
         19 . The method of  claim 18 , wherein the vector is delivered to essentially all nervous system cells of the mammal. 
     
     
         20 . The method of  claim 18 , wherein the vector is specifically delivered to particular cell types or regions of the nervous system of the mammal. 
     
     
         21 . The method of  claim 1 , wherein said method for delivering nucleic acid encoding neuropeptide Y to cells of the nervous system to effect expression of neuropeptide Y in cells of the nervous system treats a disorder of the nervous system. 
     
     
         22 . The method of  claim 21 , wherein said disorder of the nervous system is epilepsy. 
     
     
         23 . The method of  claim 22 , wherein said epilepsy is intractable epilepsy. 
     
     
         24 . The method of  claim 23 , wherein said epilepsy is temporal lobe epilepsy. 
     
     
         25 . The method of  claim 1 , wherein said nucleic acid sequence encoding neuropeptide Y is a nucleic acid sequence encoding an amino acid sequence comprising SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, or SEQ ID NO: 8, or a derivative or functional fragment thereof. 
     
     
         26 . The method of  claim 1 , wherein said nucleic acid sequence encoding neuropeptide Y is a nucleic acid sequence encoding an amino acid sequence comprising SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, or SEQ ID NO: 8, or an amino acid sequence at least 90% homologous to SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, or SEQ ID NO: 8. 
     
     
         27 . The method of  claim 1 , wherein said nucleic acid sequence encoding neuropeptide Y is a nucleic acid sequence encoding an amino acid sequence comprising SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, or SEQ ID NO: 8, or an amino acid sequence at least 85% homologous to SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, or SEQ ID NO: 8. 
     
     
         28 . The method of  claim 1 , wherein said nucleic acid sequence encoding neuropeptide Y is a nucleic acid sequence comprising SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, or SEQ ID NO: 7, or a derivative or functional fragment thereof. 
     
     
         29 . The method of  claim 1 , wherein said nucleic acid sequence encoding neuropeptide Y is a nucleic acid sequence comprising SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, or SEQ ID NO: 7, or a nucleic acid sequence at least 90% homologous to SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, or SEQ ID NO: 7. 
     
     
         30 . The method of  claim 1 , wherein said nucleic acid sequence encoding neuropeptide Y is a nucleic acid sequence comprising SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, or SEQ ID NO: 7, or a nucleic acid sequence at least 85% homologous to SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, or SEQ ID NO: 7. 
     
     
         31 . The method of  claim 18 , wherein said administering is by stereotaxic microinjection. 
     
     
         32 . An AAV vector which retains only the replication and packaging signals of AAV, and which comprises a nucleic acid sequence encoding neuropeptide Y, or a derivative or a functional fragment thereof. 
     
     
         33 . The AAV vector of  claim 32 , wherein said nucleic acid sequence comprises a nucleic acid sequence of SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, or SEQ ID NO: 7, or a derivative or a functional fragment thereof. 
     
     
         34 . The AAV vector of  claim 32 , wherein said nucleic acid sequence encodes an amino acid sequence comprising SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, or SEQ ID NO: 8, or a derivative or a functional fragment thereof. 
     
     
         35 . A composition comprising an AAV vector of  claim 32  and a pharmaceutically acceptable carrier. 
     
     
         36 . A composition comprising an AAV vector of  claim 33  and a pharmaceutically acceptable carrier. 
     
     
         37 . A composition comprising an AAV vector of  claim 34  and a pharmaceutically acceptable carrier. 
     
     
         38 . A method for treating a mammal with a neurological disease, said method comprising administering an expression vector to a target cell in the mammal, wherein said expression vector comprises a nucleic acid sequence encoding neuropeptide Y, or a derivative or functional fragment thereof, and wherein said administering results in expression of neuropeptide Y, or a derivative or functional fragment thereof, in said target cell and said expression reduces the symptoms of the neurological disease, thereby treating the mammal with the neurological disease. 
     
     
         39 . The method of  claim 38 , wherein the expression vector is a viral or a non-viral expression vector. 
     
     
         40 . The method of  claim 39 , wherein the viral expression vector is an adeno-associated virus (AAV) vector, a lentivirus vector, an adenovirus vector, or a herpes simplex virus (HSV) vector. 
     
     
         41 . The method of  claim 38 , wherein said nucleic acid sequence encoding neuropeptide Y is a nucleic acid sequence encoding an amino acid sequence comprising SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, or SEQ ID NO: 8, or a derivative or a functional fragment thereof. 
     
     
         42 . The method of  claim 41 , wherein said nucleic acid sequence encoding neuropeptide Y is a nucleic acid sequence encoding an amino acid sequence comprising SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, or SEQ ID NO: 8, or an amino acid sequence at least 90% homologous to SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, or SEQ ID NO: 8. 
     
     
         43 . The method of  claim 41 , wherein said nucleic acid sequence encoding neuropeptide Y is a nucleic acid sequence encoding an amino acid sequence comprising SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, or SEQ ID NO: 8, or an amino acid sequence at least 85% homologous to SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, or SEQ ID NO: 8. 
     
     
         44 . The method of  claim 38 , wherein said nucleic acid sequence encoding neuropeptide Y is a nucleic acid sequence comprising SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, or SEQ ID NO: 7, or a derivative or a functional fragment thereof. 
     
     
         45 . The method of  claim 38 , wherein said nucleic acid sequence encoding neuropeptide Y is a nucleic acid sequence comprising SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, or SEQ ID NO: 7, or a nucleic acid sequence at least 90% homologous to SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, or SEQ ID NO: 7. 
     
     
         46 . The method of  claim 38 , wherein said nucleic acid sequence encoding neuropeptide Y is a nucleic acid sequence comprising SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, or SEQ ID NO: 7, or a nucleic acid sequence at least 85% homologous to SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, or SEQ ID NO: 7. 
     
     
         47 . The method of  claim 38 , wherein said neurological disease is epilepsy. 
     
     
         48 . The method of  claim 38 , wherein said neurological disease is intractable epilepsy. 
     
     
         49 . The method of  claim 38 , wherein said neurological disease is temporal lobe epilepsy. 
     
     
         50 . The method of  claim 38 , wherein said administering is by stereotaxic microinjection. 
     
     
         51 . The method of  claim 38 , wherein said administering is by stereotaxic microinjection to a medial temporal lobe or temporal cortex of the central nervous system. 
     
     
         52 . The method of  claim 51 , wherein said administering to the medial temporal lobe is localized to the hippocampus and/or amygdala. 
     
     
         53 . A method for delivering a nucleic acid sequence to a mammalian nervous system target cell, wherein said nucleic acid sequence is expressible in the target cell for greater than three months, said method comprising administering an adeno-associated virus (AAV) vector to the target cell, wherein said vector transduces the target cell; and wherein said AAV vector comprises an AAV vector of  claim 32 , and is free of both wildtype and helper virus. 
     
     
         54 . A method for delivering a nucleic acid sequence to a mammalian nervous system target cell, wherein said nucleic acid sequence is expressible in the target cell for greater than three months, said method comprising administering a composition of  claim 35  to the target cell, said composition comprising an AAV vector capable of transducing the target cell; and wherein said AAV vector is free of both wildtype and helper virus. 
     
     
         55 . A method for treating a mammal with a neurological disease, said method comprising administering an adeno-associated virus (AAV) vector to a target cell in the mammal, wherein said AAV vector comprises an AAV vector of  claim 32 , and wherein said administering results in expression of neuropeptide Y, or a derivative or functional fragment thereof, in said target cell and said expression reduces the symptoms of the neurological disease, thereby treating the mammal with the neurological disease. 
     
     
         56 . A method for treating a mammal with a neurological disease, said method comprising administering a composition of  claim 35  to the target cell, said composition comprising an AAV vector capable of transducing the target cell and wherein said administering results in expression of neuropeptide Y, or a derivative or functional fragment thereof, in said target cell and said expression reduces the symptoms of the neurological disease, thereby treating the mammal with the neurological disease.