Chimeric pro-caspases and methods of using same

Abstract

The present invention relates to a chimeric pro-caspase, which contains a pro-caspase domain and an oligomerizing domain. The invention also relates to an antibody that reacts specifically with a chimeric pro-caspase. In addition, the invention further relates to a polynucleotide encoding a chimeric pro-caspase, and to nucleotide sequences, which can hybridize specifically with a polynucleotide encoding a chimeric pro-caspase. The present invention also relates to a method of inducing apoptosis in a cell by providing a chimeric pro-caspase in the cell, wherein the chimeric pro-caspase includes a pro-caspase domain and an oligomerizing domain, whereby the chimeric pro-caspase forms an oligomer in the cell, thereby activating caspase activity of the chimeric pro-caspase and inducing apoptosis in the cell. The present invention further relates to a method of reducing the severity of a pathologic condition in a subject, by providing cells of the subject that are involved in the pathologic condition with a chimeric pro-caspase comprising a pro-caspase domain and an oligomerizing domain, whereby the chimeric pro-caspase forms an oligomer in the cells, thereby activating caspase activity of the chimeric pro-caspase, inducing apoptosis in the cells, and reducing the severity of the pathologic condition in the subject.

1 . A chimeric pro-caspase, comprising a pro-caspase domain and an oligomerizing domain.  
   
   
       2 . The chimeric pro-caspase of  claim 1 , wherein the pro-caspase domain comprises pro-caspase-8 or a peptide portion of said pro-caspase-8 having caspase-8 activity potential.  
   
   
       3 . The chimeric pro-caspase of  claim 1 , wherein the pro-caspase domain comprises a pro-caspase form of an initiator caspase or a peptide portion of said pro-caspase having initiator caspase activity potential.  
   
   
       4 . The chimeric pro-caspase of  claim 1 , wherein the oligomerizing domain comprises an FK506 binding protein.  
   
   
       5 . The chimeric pro-caspase of  claim 1 , wherein the oligomerizing domain comprises a polypeptide that interacts specifically with cellular protein in the cell.  
   
   
       6 . The chimeric pro-caspase of  claim 5 , wherein the oligomerizing domain comprises a Raf domain, which interacts specifically with a Ras cellular protein.  
   
   
       7 . The chimeric pro-caspase of  claim 5 , wherein the oligomerizing domain comprises a guanine exchange factor domain, which interacts specifically with a Ras cellular protein.  
   
   
       8 . The chimeric pro-caspase of  claim 1 , further comprising a protein transduction domain.  
   
   
       9 . The chimeric pro-caspase of  claim 8 , wherein the protein transduction domain is the human immunodeficiency virus TAT protein transduction domain.  
   
   
       10 . The chimeric pro-caspase of  claim 1 , further comprising a cell compartment localization domain.  
   
   
       11 . A pharmaceutical composition, comprising a chimeric pro-caspase of  claim 1 .  
   
   
       12 . An antibody that reacts specifically with a chimeric pro-caspase of  claim 1 .  
   
   
       13 . A kit comprising an antibody of  claim 12 .  
   
   
       14 . A polynucleotide encoding a chimeric pro-caspase of  claim 1 .  
   
   
       15 . The polynucleotide of  claim 14 , which is contained in a vector.  
   
   
       16 . The polynucleotide of  claim 15 , wherein the vector is an expression vector.  
   
   
       17 . The polynucleotide of  claim 16 , wherein the expression vector is a viral vector.  
   
   
       18 . A host cell containing the vector of  claim 15 .  
   
   
       19 . An oligonucleotide, which hybridizes specifically with a polynucleotide of  claim 14 .  
   
   
       20 . A pharmaceutical composition comprising the polynucleotide of  claim 14 .  
   
   
       21 . A method of inducing apoptosis in a cell, comprising providing a chimeric pro-caspase in the cell, wherein the chimeric pro-caspase comprises a pro-caspase domain and an oligomerizing domain, and whereby the chimeric pro-caspase forms an oligomer in the cell, thereby activating caspase activity of the chimeric pro-caspase and inducing apoptosis in the cell.  
   
   
       22 . The method of  claim 21 , wherein providing the chimeric pro-caspase comprises introducing a polynucleotide encoding the chimeric pro-caspase into the cell, and expressing the encoded chimeric pro-caspase.  
   
   
       23 . The method of  claim 22 , wherein the polynucleotide encoding the chimeric pro-caspase is contained in a vector.  
   
   
       24 . The method of  claim 23 , wherein the vector is a viral vector.  
   
   
       25 . The method of  claim 21 , wherein the chimeric pro-caspase further comprises a protein transduction domain, and wherein providing the chimeric pro-caspase comprises contacting the cell with the chimeric pro-caspase comprising the protein transduction domain.  
   
   
       26 . The method of  claim 25 , wherein the protein transduction domain is the human immunodeficiency virus TAT protein transduction domain.  
   
   
       27 . The method of  claim 21 , wherein the pro-caspase domain comprises a pro-caspase form of an initiator caspase or a peptide portion of said pro-caspase having initiator caspase activity potential.  
   
   
       28 . The method of  claim 21 , wherein the pro-caspase domain comprises pro-caspase-8 or a peptide portion of said pro-caspase-8 having caspase-8 activity potential.  
   
   
       29 . The method of  claim 21 , wherein the chimeric pro-caspase is induced to form an oligomer in the cell.  
   
   
       30 . The method of  claim 29 , wherein the oligomerizing domain of the chimeric pro-caspase comprises an FK506 binding protein, and wherein the chimeric pro-caspase is induced to form an oligomer due to contacting the cell with an agent that induces oligomerization of FK506 binding proteins.  
   
   
       31 - 45 . (canceled)