I-FLICE, a novel inhibitor of tumor necrosis factor receptor-1 and CD-95 induced apoptosis

Abstract

The present invention relates to a novel I-FLICE-1 or I-FLICE-2 protein which is a novel inhibitor of TNFR-1 and CD-95 induced apoptosis. In particular, isolated nucleic acid molecules are provided encoding the human I-FLICE-1 or I-FLICE-2 protein. I-FLICE-1 or I-FLICE-2 polypeptides are also provided as are vectors, host cells and recombinant methods for producing the same. The invention further relates to screening methods for identifying agonists and antagonists of I-FLICE-1 or I-FLICE-2 activity. Also provided are therapeutic methods for treating diseases and disorders associated with apoptosis.

1 . An isolated nucleic acid molecule comprising a polynucleotide having a nucleotide sequence at least 95% identical to a sequence selected from the group consisting of: 
 (a) a nucleotide sequence encoding a polypeptide comprising amino acids from about 1 to about 480 in SEQ ID NO:2;    (b) a nucleotide sequence encoding a polypeptide comprising amino acids from about 2 to about 480 in SEQ ID NO:2;    (c) a nucleotide sequence encoding a polypeptide comprising amino acids from about 1 to about 75 in SEQ ID NO:2;    (d) a nucleotide sequence encoding a polypeptide comprising amino acids from about 91 to about 171 in SEQ ID NO:2;    (e) a nucleotide sequence encoding a polypeptide comprising amino acids from about 172 to about 375 in SEQ ID NO:2;    (f) a nucleotide sequence encoding a polypeptide comprising amino acids from about 376 to about 480 in SEQ ID NO:2;    (g) a nucleotide sequence encoding a polypeptide comprising amino acids from about 1 to about 348 in SEQ ID NO:6;    (h) a nucleotide sequence encoding a polypeptide comprising amino acids from about 2 to about 348 in SEQ ID NO:6;    (i) a nucleotide sequence encoding a polypeptide comprising amino acids from about 1 to about 75 in SEQ ID NO:6;    (j) a nucleotide sequence encoding a polypeptide comprising amino acids from about 76 to about 252 in SEQ ID NO:6;    (k) a nucleotide sequence encoding a polypeptide comprising amino acids from about 253 to about 348 in SEQ ID NO:6;    (l) a nucleotide sequence encoding a polypeptide having the amnino acid sequence encoded by the cDNA clone contained in ATCC Deposit No. 209041;    (m) a nucleotide sequence encoding a polypeptide having the amino acid sequence encoded by the cDNA clone contained in ATCC Deposit No. 209038; and    (n) a nucleotide sequence complementary to any of the nucleotide sequences in (a), (b), (c), (d), (e), (f), (g), (h), (i), (j), (k), (l), (m), or (n).    
     
     
         2 . An isolated nucleic acid molecule comprising a polynucleotide which hybridizes under stringent hybridization conditions to a polynucleotide having a nucleotide sequence identical to a nucleotide sequence in (a), (b), (c), (d), (e), (f), (g), (h), (i), (j), (k), (l), (m), (n), or (o) of  claim 1  wherein said polynucleotide which hybridizes does not hybridize under stringent hybridization conditions to a polynucleotide having a nucleotide sequence consisting of only A residues or of only T residues.  
     
     
         3 . A method for making a recombinant vector comprising inserting an isolated nucleic acid molecule of  claim 1  into a vector.  
     
     
         4 . A recombinant vector produced by the method of  claim 3 .  
     
     
         5 . A method of making a recombinant host cell comprising introducing the recombinant vector of  claim 4  into a host cell.  
     
     
         6 . A recombinant host cell produced by the method of  claim 5 .  
     
     
         7 . A recombinant method for producing an I-FLICE-1 or I-FLICE-2 polypeptide, comprising culturing the recombinant host cell of  claim 6  under conditions such that said polypeptide is expressed and recovering said polypeptide.  
     
     
         8 . An isolated I-FLICE-1 or I-FLICE-2 polypeptide having an amino acid sequence at least 95% identical to a sequence selected from the group consisting of: 
 (a) amino acids from about 1 to about 480 in SEQ ID NO:2;    (b) amino acids from about 2 to about 480 in SEQ ID NO:2;    (c) amino acids from about 1 to about 348 in SEQ ID NO:6;    (d) amino acids from about 2 to about 348 in SEQ ID NO:6;    (e) the amino acid sequence of the I-FLICE-1 polypeptide having the amino acid sequence encoded by the cDNA clone contained in ATCC Deposit No. 209041;    (f) the amino acid sequence of the I-FLICE-2 polypeptide having the amino acid sequence encoded by the cDNA clone contained in ATCC Deposit No. 209038; and    (g) the amino acid sequence of an epitope-bearing portion of any one of the polypeptides of (a), (b), (c), (d), (e), or (f).    
     
     
         9 . An isolated antibody that binds specifically to an I-FLICE-1 or I-FLICE-2 polypeptide of  claim 8 .  
     
     
         10 . A method for treating diseases and disorders associated with apoptosis comprising administering to said individual a composition comprising an isolated polypeptide of  claim 8 .  
     
     
         11 . A method useful during the diagnosis of diseases and disorders associated with aberrant cell survival in an individual comprising: 
 (a) measuring I-FLICE-1 or I-FLICE-2 gene expression level in cells or body fluid of said individual;    (b) comparing the I-FLICE-1 or I-FLICE-2 gene expression level of said individual with a standard I-FLICE-1 or I-FLICE-2 gene expression level, whereby an increase or decrease in the I-FLICE-1 or I-FLICE-2 gene expression level of said individual compared to said standard expression level is indicative of disease or disorder associated with aberrant cell survival.