Survivin, a protein that inhibits cellular apoptosis, and its modulation

Abstract

The present invention provides the amino acid of a protein that inhibits cellular apoptosis, herein termed the Survivin protein and nucleic acid molecules that encode Survivin. Based on this disclosure, the present invention provides isolated Survivin protein, isolated Survivin encoding nucleic acid molecules, methods of isolating other members of the Survivin family of proteins, methods for identifying agents that block Survivin mediated inhibition of cellular apoptosis, methods of using agents that block Survivin mediated inhibition or Survivin expression to modulate biological and pathological processes, and methods of assaying Survivin activity.

1 - 59 . (canceled)  
     
     
         60 . A method of assessing the proliferative potential of a human cell, the method comprising assessing expression of survivin in the cell, whereby a greater degree of survivin expression is correlated with increased proliferative potential.  
     
     
         61 . The method of  claim 60 , wherein expression of survivin in the cell is assessed by assessing occurrence of survivin shed from the cell into a surrounding biological fluid.  
     
     
         62 . The method of  claim 61 , wherein the fluid is selected from the group consisting of blood, plasma, semen, urine, and saliva.  
     
     
         63 . The method of  claim 60 , wherein expression of survivin in the cell is assessed by assessing occurrence of survivin in an extract prepared from the cell.  
     
     
         64 . The method of  claim 60 , wherein the cell is a tumor cell.  
     
     
         65 . The method of  claim 60 , wherein the cell is selected from the group consisting of a lung cell, a colon cell, a breast cell, a pancreas cell, a prostate cell, and a lymphocyte.  
     
     
         66 . The method of  claim 60 , wherein survivin expression is assessed by an immunological method.  
     
     
         67 . The method of  claim 66 , comprising contacting an extract of the cell with a first antibody that specifically binds with survivin.  
     
     
         68 . The method of  claim 67 , wherein the antibody is a monoclonal antibody.  
     
     
         69 . The method of  claim 67 , further comprising contacting any complex formed by binding of the first antibody and survivin with a second antibody that binds with the complex.  
     
     
         70 . The method of  claim 66 , comprising contacting an extract of the cell with a fragment of an antibody that specifically binds with survivin, wherein the fragment includes the survivin-binding domain of the antibody.  
     
     
         71 . The method of  claim 66 , wherein survivin expression is assessed in a biological fluid that contacts the cell.  
     
     
         72 . The method of  claim 60 , wherein survivin expression is assessed by assessing occurrence of mRNA encoding survivin in the cell.  
     
     
         73 . The method of  claim 72 , wherein occurrence of the mRNA is assessed by contacting an extract of the cell with a single-stranded polynucleotide probe that hybridizes under stringent conditions with mRNA encoding survivin, wherein the stringent conditions are selected from the group consisting of: (1) washing with 0.015 M sodium chloride, 0.0015 M sodium titrate, 0.1% SDS at 50° C.; (2) hybridization in 50% (vol/vol) formamide with 0.1% bovine serum albumin, 0.1% Ficoll, 0.1% polyvinylpyrrolidone, 50 mM sodium phosphate buffer at pH 6.5 with 750 mM sodium chloride, 75 mM sodium citrate at 42° C.; and (3) hybridization in 50% formamide, 5×SSC (0.75 M sodium chloride, 0.075 M sodium citrate), 50 mM sodium phosphate (pH 6.8), 0.1% sodium pyrophosphate, 5× Denhardt's solution, sonicated salmon sperm DNA (50 micrograms/milliliter), 0.1% SDS, and 10% dextran sulfate at 42° C., with washes at 42° C. in 0.2×SSC and 0.1% SDS.  
     
     
         74 . The method of  claim 73 , further comprising amplifying any double-stranded nucleic acid formed by hybridization of the probe and the mRNA.  
     
     
         75 . The method of  claim 73 , further comprising extending the probe and amplifying any double-stranded nucleic acid formed by hybridization of the probe and the mRNA.  
     
     
         76 . The method of  claim 73 , wherein the probe hybridizes under stringent conditions with mRNA encoding survivin but not with a polynucleotide having the sequence SEQ ID NO: 35.  
     
     
         77 . A method of assessing the cancerous state of a tissue, the method comprising assessing the proliferative potential of at least one cell obtained from the tissue by the method of  claim 60 , whereby increased proliferative potential of the cell, relative to normal cells of the same tissue, is an indication that the tissue is cancerous.  
     
     
         78 . The method of  claim 77 , wherein the tissue is selected from the group consisting of lung, colon, breast, pancreas, and prostate tissues.  
     
     
         79 . The method of  claim 77 , wherein the tissue is blood and the cell is a lymphocyte.