Methods of screening for modulators of cell proliferation and methods of diagnosing cell proliferation states

Abstract

Described herein are methods that can be used for diagnosis and prognosis of cellular proliferation. Also described herein are methods that can be used to screen candidate bioactive agents for the ability to modulate cellular proliferation. Additionally, methods and molecular targets (genes and their products) for therapeutic intervention in cancers are described.

1 . A method of screening drug candidates comprising: 
 (a) providing a cell that expresses recombinant human KSP or a fragment thereof;    (b) adding a drug candidate to said cell under conditions where the drug candidate is taken up by the cell; and    (c) determining the effect of said drug candidate on the bioactivity of said recombinant human KSP.    
     
     
         2 - 59 . (Canceled)  
     
     
         60 . A method of assessing an individual's risk for a hyper-proliferative disorder, comprising: 
 (a) determining the expression level of KSP in a sample obtained from the individual; and    (b) comparing the KSP expression level in the sample with the expression level of KSP in a control of known proliferation state; and    (c) assessing the individual's risk for the hyper-proliferative disorder on the basis of the comparison of step (b).    
     
     
         61 . The method of  claim 60 , wherein 
 comparing comprises comparing the KSP expression level in the sample with a control that is representative of normal cells not in a hyper-proliferative state;    and assessing comprises identifying the individual as at risk for the hyper-proliferative disorder if there is a difference in KSP expression levels between the sample and the control.    
     
     
         62 . The method of  claim 60 , wherein 
 determining comprises determining the expression level of a plurality of target molecules correlated with cell proliferation, wherein one of the target molecules is KSP;    comparing comprises comparing the expression levels of each of the plurality of target molecules with the expression level of the same target molecules in the control.    
     
     
         63 . The method of  claim 62 , wherein the plurality of target molecules include a plurality of kinesins.  
     
     
         64 . The method of  claim 60 , wherein determining comprises determining the amount of nucleic acid encoding KSP.  
     
     
         65 . The method of  claim 64 , wherein the amount of nucleic acid encoding KSP is determined by the extent of binding to probes of a nucleic acid probe array that specifically hybridize to nucleic acids encoding KSP.  
     
     
         66 . The method of  claim 64 , wherein the amount of nucleic acid encoding KSP is determined by in situ hybridization.  
     
     
         67 . The method of  claim 64 , wherein the nucleic acid is DNA.  
     
     
         68 . The method of  claim 64 , wherein the nucleic acid is RNA.  
     
     
         69 . The method of  claim 60 , wherein determining comprises determining the amount of KSP protein in the sample.  
     
     
         70 . The method of  claim 69 , wherein the amount of KSP protein is determined by mass spectroscopy.  
     
     
         71 . The method of  claim 69 , wherein the amount of KSP protein is determined by an immunological method.  
     
     
         72 . The method of  claim 70 , wherein the immunological method is an enzyme-linked immunoassay assay (ELISA).  
     
     
         73 . The method of  claim 69 , wherein the KSP protein level is determined by two-dimensional gel electrophoresis.  
     
     
         74 . The method of  claim 60 , wherein the hyper-proliferative disorder is a cancer.  
     
     
         75 . The method of  claim 60 , further comprising determining whether a variant form of a cell proliferation gene is present in the sample, the presence of a variant cell proliferation gene being an indication that the individual is at risk for the hyper-proliferative disorder.  
     
     
         76 . The method of  claim 75 , wherein the cell proliferation gene is KSP.  
     
     
         77 . The method of  claim 61 , wherein 
 the hyper-proliferative disorder is a cancer; and    determining comprises determining the amount of nucleic acid encoding KSP from the extent of binding to probes of a nucleic acid probe array that specifically hybridize to nucleic acids encoding KSP or by in situ hybridization.