Methods for the prognostic and/or diagnostic of neurodegenerative disease, methods to identify candidate compounds and compounds for treating neurodegenerative disease

Abstract

The present invention provides methods and uses for the diagnostic of a subject predisposed or suspected of developing a neurodegenerative disease or suffering from a neurodegenerative disease. The present invention also relates to methods and uses for identifying candidate compounds and to compounds for treating neurodegenerative disease. The present invention also relates to an animal model for neurodegenerative disease.

1 - 80 . (canceled) 
     
     
         81 . A method for the diagnostic of a subject predisposed or suspected of developing a neurodegenerative disease or suffering from a neurodegenerative disease, the method comprising the steps of:
 contacting a TDP-43 polypeptide or fragment thereof with a TDP-43 agent in a biological sample of the subject;   contacting a p65 polypeptide or fragment thereof with a p65 agent in the biological sample; and   detecting the TDP-43 agent and/or the p65 agent to determine the level of interaction between the TDP-43 polypeptide or fragment thereof and the p65 polypeptide or fragment thereof,   wherein detecting an elevated level of interaction between TDP-43 polypeptide or fragment thereof and p65 polypeptide or fragment thereof in the biological sample relative to a reference level of interaction between TDP-43 polypeptide or fragment thereof and p65 polypeptide or fragment thereof indicates that the subject is predisposed or suspected of developing a neurodegenerative disease or is suffering from a neurodegenerative disease.   
     
     
         82 . The method of  claim 81 , wherein the elevated level of interaction between TDP-43 polypeptide or fragment thereof and p65 polypeptide or fragment thereof is at least 1.5-fold higher than the reference. 
     
     
         83 . The method of  claim 81 , wherein p65 polypeptide or fragment thereof interacts with the N-terminal portion of TDP-43 polypeptide or fragment thereof. 
     
     
         84 . The method of  claim 81 , wherein p65 polypeptide or fragment thereof interacts with a RRM domain of TDP-43 or fragment thereof. 
     
     
         85 . The method of  claim 81 , wherein the level of interaction between TDP-43 polypeptide or fragment thereof and p65 polypeptide or fragment thereof is determined by ELISA, radioimmunoassay, immunoprecipitation assay, Western blot assay, EMSA supershift or immunostaining assay. 
     
     
         86 . The method of  claim 81 , wherein the biological sample is from blood, urine, cerebrospinal fluid, cortical neurons, microglial cells, myeloid cells or spinal cord extract. 
     
     
         87 . The method of  claim 81  wherein the neurodegenerative disease is amyotropic lateral sclerosis, frontotemporal lobar degeneration, Alzheimer, motor neuron disease or Parkinson. 
     
     
         88 . The method of  claim 81  wherein the neurodegenerative disease is associated with TDP-43 proteinopathy. 
     
     
         89 . The method of  claim 81  wherein the determination of the level of interaction between TDP-43 polypeptide or fragment thereof and p65 polypeptide is repeated after a time interval in order to monitor the progression or regression of the neurodegenerative disease, and wherein detecting an increased level of interaction between TDP-43 polypeptide or fragment thereof and p65 polypeptide or fragment thereof over the time interval indicates a progression of the neurodegenerative disease and wherein observing a decreased level of interaction between TDP-43 polypeptide or fragment thereof and p65 polypeptide or fragment thereof over the time interval indicates a regression of the neurodegenerative disease. 
     
     
         90 . The method of  claim 89 , wherein the time interval is hourly, daily weekly, monthly or yearly. 
     
     
         91 . A kit for the diagnostic of a subject predisposed to or suspected of developing a neurodegenerative disease or suffering from a neurodegenerative disease for use with a biological sample of the subject, the kit comprising:
 i) at least one TDP-43 specific antibody or fragment thereof;   ii) at least one p65 specific antibody or fragment thereof;   iii) a reference corresponding to the level of interaction between TDP-43 polypeptide or fragment thereof and p65 polypeptide or fragment thereof,   iv) a container, and   v) a buffer or an appropriate reagent.   
     
     
         92 . The kit of  claim 91 , wherein the specific TDP-43 antibody and the specific p65 antibody are purified monoclonal or purified polyclonal antibodies or fragments thereof. 
     
     
         93 . The kit of  claim 91 , wherein the biological sample is from blood, urine, cerebrospinal fluid, cortical neurons, microglial cells, myeloid cells or spinal cord extract. 
     
     
         94 . A method for identifying a candidate compound useful for preventing and/or treating a neurodegenerative disease, the method comprising the steps of:
 contacting the candidate compound with a biological system comprising TDP-43 polypeptide or fragment thereof and p65 polypeptide or fragment thereof, and
 a) measuring the ability of the candidate compound to reduce or inhibit the interaction between TDP-43 polypeptide or fragment thereof and p65 polypeptide or fragment thereof, and/or 
 b) measuring the ability of the candidate compound to modulate the activation of NF-κB p65 in the biological system, and 
   determining if the candidate compound is useful for preventing and/or treating a neurodegenerative disease based on the result of step a) or step b) or the results of step a) and step b).   
     
     
         95 . The method of  claim 94 , wherein the measuring step a) comprises an ELISA assay, immunoprecipitation assay, coimmunoprecipitation assay, Western Blot assay, immunostaining assay or radioimmunoassay. 
     
     
         96 . A method for treating a subject suffering from a neurodegenerative disease comprising the step of administering at least one withanolide compound or pharmaceutically acceptable salt thereof to the subject. 
     
     
         97 . The method of  claim 96 , wherein the withanolide compound is withaferin A, withanolide N, withanolide O, withanolide D, withanolide E, withanolide P, withanolide S, withanolide Q, withanolide R, withanolide G, withanolide H, withanolide I, withanolide J, withanolide K, withanolide U, withanolide Y, analogs thereof or pharmaceutically salt thereof. 
     
     
         98 . The method of  claim 96 , wherein the withanolide compound is withaferin A, an analog or a pharmaceutically acceptable salt thereof. 
     
     
         99 . The method of  claim 96 , wherein the neurodegenerative disease is amyotropic lateral sclerosis, frontotemporal lobar degeneration, Alzheimer, motor neuron disease or Parkinson. 
     
     
         100 . The method of  claim 96 , wherein the neurodegenerative disease is associated with TDP-43 proteinopathy.