Novel Gene Targets Associated with Amyotrophic Lateral Sclerosis and Methods of Use Thereof

Abstract

Novel gene targets and methods of use thereof for the diagnosis and treatment of Amyotrophic Lateral Sclerosis are provided.

1 . A method for predicting an increased risk of an individual for developing amyotrophic lateral sclerosis (ALS) disease, said method comprising: obtaining a nucleic acid sample encoding EWSR1 from said individual and determining whether or not said EWSR1 comprises a mutation selected from the group consisting of
 i) a missense mutation in exon 16 at 1532 G>C causing a Gly511Ala alteration, and   ii) a missense mutation in exon 17 at 1655 C>T causing a Pro522Leu alteration, relative to wild-type EWSR1 encoding nucleic acids, wherein the presence of at least one mutation in said EWSR1 encoding nucleic acid when compared to wild type EWSR1 encoding nucleic acids obtained from patients not having ALS is indicative of an increased risk of ALS.   
     
     
         2 . (canceled) 
     
     
         3 . The method of  claim 1 , wherein said method predicts an increased risk of early onset ALS. 
     
     
         4 . The method of  claim 1 , wherein said nucleic acid is obtained from a blood, tissue or skin sample. 
     
     
         5 . The method of  claim 1 , wherein said mutations are detected using an EWSR1 specific probe or primer. 
     
     
         6 . A diagnostic kit for performing the method of  claim 5 , comprising reagents suitable for isolation of DNA, and reagents suitable for detection of said mutation containing EWSR1 nucleic acids. 
     
     
         7 . A method for identifying agents which inhibit TDP-43 associated complex formation with at least one RNA binding protein listed in Table 1, comprising:
 a) providing a cell which expresses TDP-43 and said RNA binding protein, said expression being associated with increased cellular toxicity and cytoplasmic aggregate formation;   b) contacting said cell with an effective amount of an agent; and   c) measuring cellular toxicity and/or aggregate formation in the presence of said agent relative to a non-treated control cell, wherein a decrease in cellular toxicity identifies an agent which reduces TDP-43-RNA binding protein mediated cellular toxicity and cytoplasmic aggregate formation.   
     
     
         8 . The method of  claim 7 , wherein said cell is a  Saccharomyces cerevisiae  or  Drosophila  cell. 
     
     
         9 . The method of  claim 7 , wherein said RNA binding protein is EWSR1, said EWSR1 optionally comprising at least one missense mutation. 
     
     
         10 . The method of  claim 7 , wherein said RNA binding protein is TAF15, said TAF15 optionally comprising at least one missense mutation. 
     
     
         11 . An agent identified via the method of  claim 8  or  claim 9 . 
     
     
         12 . An isolated nucleic acid encoding an altered EWSR1 protein comprising a genetic alteration selected from the group consisting of
 i) a missense mutation in exon 16 at 1532 G>C causing a Gly511Ala alteration, and   ii) a missense mutation in exon 17 at 1655 C>T causing a Pro522Leu alteration.   
     
     
         13 . An isolated altered EWSR1 protein encoded by the nucleic acid of  claim 12 . 
     
     
         14 . (canceled) 
     
     
         15 . (canceled) 
     
     
         16 . The nucleic acid of  claim 12  affixed to a solid support. 
     
     
         17 . A vector comprising the nucleic acid of  claim 12 . 
     
     
         18 . A host cell comprising the vector of  claim 17 . 
     
     
         19 . The host cell of  claim 18  which is a  drosophila  cell or a yeast cell.