JNK3 modulators and methods of use

Abstract

The c-Jun NH 2 -terminal kinase (JNK) group of MAP kinases are activated by exposure of cells to environmental stress. The role of JNK in the brain was examined by targeted disruption of the gene that encodes the neuronal isoform JNK3. It was found that JNK3 is required for the normal response to seizure activity. Methods of screening for molecules and compounds that decrease JNK3 expression or activity are described. Such molecules or compounds are useful for treating disorders involving excitotoxicity such as seizure disorders, Alzheimer's disease, Huntington disease, Parkinson's disease, and ischaemia.

1 - 17 . (canceled)  
     
     
         18 . A method for identifying a modulator of c-Jun N-terminal kinase (JNK3), the method comprising: 
 (a) exposing a cell to a stress signal in the presence of a test compound under conditions that allow the cell to phosphorylate c-Jun; and    (b) determining the level of phosphorylated c-Jun in the cell,    wherein a difference in the level of phosphorylated c-Jun determined in (b) compared to the level of phosphorylated c-Jun in the absence of the test compound indicates that the test compound is a modulator of JNK3.    
     
     
         19 . The method of  claim 18 , wherein the cell is a neuronal cell.  
     
     
         20 . The method of  claim 19 , wherein the neuronal cell is a hippocampal cell.  
     
     
         21 . The method of  claim 18 , wherein the test compound is an antisense nucleic acid, an oligonucleotide, a ribozyme, a peptide, a peptidomimetic, an amino acid, a phosphopeptide, a small organic molecule, or a small inorganic molecule.  
     
     
         22 . A method for identifying a modulator of c-Jun N-terminal kinase (JNK3), the method comprising: 
 (a) exposing a cell to a stress signal in the presence of a test compound under conditions that allow the cell to increase AP-1 transcription activity; and    (b) determining the level of AP-1 transcription activity in the cell,    wherein a difference in the level of AP-1 transcription activity determined in (b) compared to the level of AP-1 transcription activity in the absence of the test compound indicates that the test compound is a modulator of JNK3.    
     
     
         23 . The method of  claim 22 , wherein the cell is a neuronal cell.  
     
     
         24 . The method of  claim 23 , wherein the neuronal cell is a hippocampal cell.  
     
     
         25 . The method of  claim 22 , wherein AP-1 transcription activity is measured using a transgene comprising an AP-1 binding site and a reporter gene.  
     
     
         26 . The method of  claim 22 , wherein the test compound is an antisense nucleic acid, an oligonucleotide, a ribozyme, a peptide, a peptidomimetic, an amino acid, a phosphopeptide, a small organic molecule, or a small inorganic molecule.  
     
     
         27 . A method of identifying a compound that modulates excitotoxicity, the method comprising: 
 (a) identifying a modulator of c-Jun N-terminal kinase (JNK3) expression;    (b) incubating a cell that can exhibit excitotoxicity with the modulator of JNK3 expression under conditions sufficient for the cell to exhibit excitotoxicity in the absence of the modulator of JNK3 expression; and    (c) determining excitotoxicity in the cell after (b),    wherein a difference in the level of excitotoxicity determined in (c) compared to the level of excitotoxicity in the absence of the modulator of JNK3 expression indicates that the modulator of JNK3 expression modulates excitotoxicity.    
     
     
         28 . The method of  claim 27 , wherein the modulator is an antisense nucleic acid, an oligonucleotide, a ribozyme, a peptide, a peptidomimetic, an amino acid, a phosphopeptide, a small organic molecule, or a small inorganic molecule.  
     
     
         29 . A method of identifying a compound that modulates excitotoxicity, the method comprising: 
 (a) identifying a modulator of c-Jun N-terminal kinase (JNK3) activity;    (b) incubating a cell that can exhibit excitotoxicity with the modulator of JNK3 activity under conditions for the cell to exhibit excitotoxicity in the absence of the modulator of JNK3 activity; and    (c) determining excitotoxicity in the cell after (b),    wherein a difference in the level of excitotoxicity determined in (c) compared to the level of excitotoxicity in the absence of the modulator of JNK3 activity indicates that the modulator of JNK3 activity modulates excitotoxicity.    
     
     
         30 . The method of  claim 29 , wherein the modulator is an oligonucleotide, a peptide, a peptidomimetic, an amino acid, a phosphopeptide, a small organic molecule, or a small inorganic molecule.  
     
     
         31 . A method of identifying a compound that modulates excitotoxicity, the method comprising: 
 (a) identifying a modulator of c-Jun N-terminal kinase (JNK3) substrate binding;    (b) incubating a cell that can exhibit excitotoxicity with the modulator of JNK3 substrate binding under conditions for the cell to exhibit excitotoxicity in the absence of the modulator of JNK3 substrate binding; and    (c) determining excitotoxicity in the cell after (b),    wherein a difference in the level of excitotoxicity determined in (c) compared to the level of excitotoxicity in the absence of the modulator of JNK3 substrate binding indicates that the modulator of JNK3 substrate binding modulates excitotoxicity.    
     
     
         32 . The method of  claim 31 , wherein the modulator is an oligonucleotide, a peptide, a peptidomimetic, an amino acid, a phosphopeptide, a small organic molecule, or a small inorganic molecule.