Methods of modulating cell death based on the bit1/aes regulatory pathway

Abstract

The present invention provides a method of identifying an effective agent that alters the association of a Bit1 polypeptide with an AES polypeptide. The method is practiced by contacting a Bit1 polypeptide, or active fragment thereof, and an AES polypeptide, or active fragment thereof, with an agent under conditions that allow the Bit1 polypeptide or active fragment thereof to associate with the AES polypeptide or active fragment thereof; and detecting an altered association of the Bit1 polypeptide or active fragment thereof and the AES polypeptide or active fragment thereof, where an altered association indicates that the agent is an effective agent that alters the association of a Bit1 polypeptide with an AES polypeptide. Such an effective agent can modulate apoptosis and can be a useful therapeutic agent.

1 - 14 . (canceled) 
     
     
         15 . A method of identifying an effective agent that modulates cell death, comprising the steps of:
 (a) contacting a cell containing an exogenous nucleic acid molecule encoding a Bit1 polypeptide or active fragment thereof with an agent; and   (b) detecting altered cell death, said altered cell death indicating that said agent is an effective agent that modulates cell death.   
     
     
         16 . The method of  claim 15 , wherein said cell expresses an AES polypeptide or active fragment thereof 
     
     
         17 . The method of  claim 16 , wherein said AES polypeptide is an exogenous AES polypeptide or active fragment thereof. 
     
     
         18 . The method of  claim 15 , wherein said cell is a mammalian cell. 
     
     
         19 . The method of  claim 15 , wherein said altered cell death is increased cell death. 
     
     
         20 . The method of  claim 15 , wherein said altered cell death is decreased cell death. 
     
     
         21 . The method of  claim 15 , wherein step (b) comprises an assay selected from the group consisting of trypan blue exclusion, thymidine uptake, deoxytransferase-mediated (TdT) dUTP biotin nick end-labeling (TUNEL), digoxygenin labeling, and a DNA filter elution assay. 
     
     
         22 . A method of identifying an effective agent that modulates apoptosis, comprising the steps of:
 (a) contacting a cell containing an exogenous nucleic acid molecule encoding a Bit1 polypeptide or active fragment thereof and a Bcl-2 promoter with an agent; and   (b) detecting an altered Bcl-2 level, said altered Bcl-2 level indicating that said agent is an effective agent that modulates apoptosis.   
     
     
         23 . The method of  claim 22 , wherein said cell expresses an AES polypeptide or active fragment thereof 
     
     
         24 . The method of  claim 23 , wherein said AES polypeptide is an exogenous AES polypeptide or active fragment thereof. 
     
     
         25 . The method of  claim 22 , wherein said altered Bcl-2 level is an increased level. 
     
     
         26 . The method of  claim 22 , wherein said altered Bcl-2 level is a decreased level. 
     
     
         27 . The method of  claim 22 , wherein said Bcl-2 promoter is operably linked to a reporter gene. 
     
     
         28 . The method of  claim 27 , wherein said reporter gene is selected from the group consisting of luciferase, green fluorescent protein (GFP) and β-galactosidase (β-GAL). 
     
     
         29 - 46 . (canceled) 
     
     
         47 . A method of preventing or reducing the severity of a disorder of cell loss in a subject, comprising administering to said subject an effective agent that selectively decreases Bit1 expression or activity, thereby inhibiting apoptosis in said subject. 
     
     
         48 . The method of  claim 47 , wherein said disorder of cell loss is selected from the group consisting of Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, retinitis pigmentosa, anemia, myocardial infarction and stroke. 
     
     
         49 . The method of  claim 47 , wherein said agent is a small molecule. 
     
     
         50 . The method of  claim 47 , wherein said agent is a nucleic acid molecule. 
     
     
         51 . The method of  claim 47 , wherein said agent is a polypeptide, peptide or peptidomimetic. 
     
     
         52 . The method of  claim 47 , wherein said agent is linked to a homing peptide. 
     
     
         53 . The method of  claim 47 , wherein said agent is an antibody or antigen-binding fragment thereof. 
     
     
         54 - 96 . (canceled)