Map3ks as modifiers of the p53 pathway and methods of use

Abstract

Human MAP3K genes are identified as modulators of the p53 pathway, and thus are therapeutic targets for disorders associated with defective p53 function. Methods for identifying modulators of p53, comprising screening for agents that modulate the activity of MAP3K are provided.

1 . A method of identifying a candidate p53 pathway modulating agent, said method comprising the steps of: 
 (a) providing an assay system comprising a purified MAP3K polypeptide or nucleic acid or a functionally active fragment or derivative thereof;    (b) contacting the assay system with a test agent under conditions whereby, but for the presence of the test agent, the system provides a reference activity; and    (c) detecting a test agent-biased activity of the assay system, wherein a difference between the test agent-biased activity and the reference activity identifies the test agent as a candidate p53 pathway modulating agent.    
     
     
         2 . The method of  claim 1  wherein the assay system comprises cultured cells that express the MAP3K polypeptide.  
     
     
         3 . The method of  claim 2  wherein the cultured cells additionally have defective p53 function.  
     
     
         4 . The method of  claim 1  wherein the assay system includes a screening assay comprising a MAP3K polypeptide, and the candidate test agent is a small molecule modulator.  
     
     
         5 . The method of  claim 4  wherein the assay is a kinase assay.  
     
     
         6 . The method of  claim 1  wherein the assay system is selected from the group consisting of an apoptosis assay system, a cell proliferation assay system, an angiogenesis assay system, and a hypoxic induction assay system.  
     
     
         7 . The method of  claim 1  wherein the assay system includes a binding assay comprising a MAP3K polypeptide and the candidate test agent is an antibody.  
     
     
         8 . The method of  claim 1  wherein the assay system includes an expression assay comprising a MAP3K nucleic acid and the candidate test agent is a nucleic acid modulator.  
     
     
         9 . The method of  claim 8  wherein the nucleic acid modulator is an antisense oligomer.  
     
     
         10 . The method of  claim 8  wherein the nucleic acid modulator is a PMO.  
     
     
         11 . The method of  claim 1  additionally comprising: 
 (d) administering the candidate p53 pathway modulating agent identified in (c) to a model system comprising cells defective in p53 function and, detecting a phenotypic change in the model system that indicates that the p53 function is restored.    
     
     
         12 . The method of  claim 11  wherein the model system is a mouse model with defective p53 function.  
     
     
         13 . A method for modulating a p53 pathway of a cell comprising contacting a cell defective in p53 function with a candidate modulator that specifically binds to a MAP3K polypeptide comprising an amino acid sequence selected from group consisting of SEQ ID NOS: 8 and 9, whereby p53 function is restored.  
     
     
         14 . The method of  claim 13  wherein the candidate modulator is administered to a vertebrate animal predetermined to have a disease or disorder resulting from a defect in p53 function.  
     
     
         15 . The method of  claim 13  wherein the candidate modulator is selected from the group consisting of an antibody and a small molecule.  
     
     
         16 . The method of  claim 1 , comprising the additional steps of: 
 (d) providing a secondary assay system comprising cultured cells or a non-human animal expressing MAP3K,    (e) contacting the secondary assay system with the test agent of (b) or an agent derived therefrom under conditions whereby, but for the presence of the test agent or agent derived therefrom, the system provides a reference activity; and    (f) detecting an agent-biased activity of the second assay system,    wherein a difference between the agent-biased activity and the reference activity of the second assay system confirms the test agent or agent derived therefrom as a candidate p53 pathway modulating agent,    and wherein the second assay detects an agent-biased change in the p53 pathway.    
     
     
         17 . The method of  claim 16  wherein the secondary assay system comprises cultured cells.  
     
     
         18 . The method of  claim 16  wherein the secondary assay system comprises a non-human animal.  
     
     
         19 . The method of  claim 18  wherein the non-human animal mis-expresses a p53 pathway gene.  
     
     
         20 . A method of modulating p53 pathway in a mammalian cell comprising contacting the cell with an agent that specifically binds a MAP3K polypeptide or nucleic acid.  
     
     
         21 . The method of  claim 20  wherein the agent is administered to a mammalian animal predetermined to have a pathology associated with the p53 pathway.  
     
     
         22 . The method of  claim 20  wherein the agent is a small molecule modulator, a nucleic acid modulator, or an antibody.  
     
     
         23 . A method for diagnosing a disease in a patient comprising: 
 (a) obtaining a biological sample from the patient;    (b) contacting the sample with a probe for MAP3K expression;    (c) comparing results from step (b) with a control;    (d) determining whether step (c) indicates a likelihood of disease.    
     
     
         24 . The method of  claim 23  wherein said disease is cancer.  
     
     
         25 . The method according to  claim 24 , wherein said cancer is a cancer as shown in Table 1 as having >25% expression level.