Inhibition of apoptosis-specific eIF-5A ("eIF-5A1") with antisense oligonucleotides and siRNA as anti-inflammatory therapeutics

Abstract

The present invention relates to apoptosis specific eucaryotic initiation factor 5A (eIF-5A), referred to as apoptosis-specific eIF-5A or eIF5A1, nucleic acids and polypeptides and methods for inhibiting or suppressing apoptosis in cells using antisense nucleotides or siRNAs to inhibit expression of apoptosis-specific eIF-5A. The invention also relates to suppressing or inhibiting expression of pro-inflammatory cytokines or inhibiting activation of NFKB by inhibiting expression of apoptosis-specific eIF-5A.

1 . An antisense oligonucleotide of apoptosis-specific eIF-5A wherein said antisense oligonucleotide suppresses endogenous expression of apoptosis-specific eIF-5A in a cell.  
     
     
         2 . The antisense oligonucleotide of  claim 1 , wherein apoptosis-specific eIF-5A is encoded by nucleotide sequence of SEQ ID NO:20.  
     
     
         3 . The antisense oligonucleotide of  claim 1 , wherein apoptosis-specific eIF-5A is encoded by nucleotide sequence of SEQ ID NO:41.  
     
     
         4 . The antisense oligonucleotide of  claim 1  wherein the antisense oligonucleotide has the sequence set forth in SEQ ID NO:35.  
     
     
         5 . The antisense oligonucleotide of  claim 1  wherein the antisense oligonucleotide has the sequence set forth in SEQ ID NO:37.  
     
     
         6 . The antisense oligonucleotide of  claim 1  wherein the antisense oligonucleotide has the sequence set forth in SEQ ID NO:39.  
     
     
         7 . An expression vector for the transfection of a mammalian cell comprising the antisense oligonucleotide of  claim 1  and regulatory sequences operatively linked to the antisense oligonucleotide to allow transcription of said antisense oligonucleotide in said cell.  
     
     
         8 . An expression vector for the transfection of a mammalian cell comprising the antisense oligonucleotide of  claim 4  and regulatory sequences operatively linked to the antisense oligonucleotide to allow transcription of said antisense oligonucleotide in said cell.  
     
     
         9 . An expression vector for the transfection of a mammalian cell comprising the antisense oligonucleotide of  claim 5  and regulatory sequences operatively linked to the antisense oligonucleotide to allow transcription of said antisense oligonucleotide in said cell.  
     
     
         10 . An expression vector for the transfection of a mammalian cell comprising the antisense oligonucleotide of  claim 6  and regulatory sequences operatively linked to the antisense oligonucleotide to allow transcription of said antisense oligonucleotide in said cell.  
     
     
         11 . A method of inhibiting expression of apoptosis-specific eIF-5A in a cell, the method comprising administering the expression vector of  claim 7  to said cell whereby said apoptosis-specific eIF-5A antisense oligonucleotide inhibits expression of endogenous apoptosis-specific eIF-5A in said cell.  
     
     
         12 . A method of inhibiting expression of apoptosis-specific eIF-5A in a cell, the method comprising administering the expression vector of  claim 8 ,  9 , or  10  to said cell whereby said apoptosis-specific eIF-5A antisense oligonucleotide inhibits expression of endogenous apoptosis-specific eIF-5A in said cell.  
     
     
         13 . The method of  claim 11  wherein said inhibition of expression of endogenous apoptosis-specific eIF-5A in said cell has an effect on the cell selected from the group consisting of suppressing apoptosis in said cell, reducing expression of p53 in said cell, reducing levels of a cytokine produced in said cell, reducing levels of a cytokine produced in said cell, increasing expression of Bcl-2 in said cell; reducing levels of myeloperoxidase produced in said cell, reducing levels of active NFk beta in said cell, reducing levels of TLR4 in said cell, reducing levels of TNFR-1 in said cell and reducing levels of iNOS in said cell.  
     
     
         14 . The method of  claim 12  herein said inhibition of expression of endogenous apoptosis-specific eIF-5A in said cell has an effect on the cell selected from the group consisting of suppressing apoptosis in said cell, reducing expression of p53 in said cell, reducing levels of a cytokine produced in said cell, reducing levels of a cytokine produced in said cell, increasing expression of Bcl-2 in said cell; reducing levels of myeloperoxidase produced in said cell, and reducing levels of active NFk beta in said cell, reducing levels of TLR4 in said cell, reducing levels of TNFR-1 in said cell and reducing levels of iNOS in said cell.  
     
     
         15 . A method delivering siRNA to lung cells of a mammal in vivo, the method comprising mixing said siRNA with water and delivering to a mammal intranasally.  
     
     
         16 . An siRNA of apoptosis-specific eIF-5A wherein said siRNA suppresses endogenous expression of apoptosis-specific eIF-5A in a cell.  
     
     
         17 . The siRNA of  claim 16 , wherein apoptosis-specific eIF-5A is encoded by nucleotide sequence of SEQ ID NO:20.  
     
     
         18 . The siRNA of  claim 16 , wherein apoptosis-specific eIF-5A is encoded by nucleotide sequence of SEQ ID NO:41.  
     
     
         19 . The siRNA of  claim 16  wherein the siRNA has the sequence set forth in SEQ ID NO:30.  
     
     
         20 . The siRNA of  claim 16  wherein the siRNA has the sequence set forth in SEQ ID NO:31.  
     
     
         21 . The siRNA of  claim 16  wherein the siRNA has the sequence set forth in SEQ ID NO:32.  
     
     
         22 . The siRNA of  claim 16  wherein the siRNA has the sequence set forth in SEQ ID NO:33.  
     
     
         23 . A method of inhibiting expression of apoptosis-specific eIF-5A in a cell, the method comprising administering the siRNA of  claim 16  to said cell whereby said apoptosis-specific eIF-5A siRNA inhibits expression of endogenous apoptosis-specific eIF-5A in said cell.  
     
     
         24 . A method of inhibiting expression of apoptosis-specific eIF-5A in a cell, the method comprising administering the siRNA of  claim 19 ,  20 ,  21 , or  22  to said cell whereby said apoptosis-specific eIF-5A siRNA inhibits expression of endogenous apoptosis-specific eIF-5A in said cell.  
     
     
         25 . A method of inhibiting expression of apoptosis-specific eIF-5A in a cell, the method comprising administering the siRNA of  claim 16  to said cell whereby said apoptosis-specific eIF-5A siRNA inhibits expression of endogenous apoptosis-specific eIF-5A in said cell.  
     
     
         26 . A method of inhibiting expression of apoptosis-specific eIF-5A in a cell, the method comprising administering the siRNA of  claim 19 ,  20 ,  21 , or  22  to said cell whereby said apoptosis-specific eIF-5A siRNA inhibits expression of endogenous apoptosis-specific eIF-5A in said cell.  
     
     
         27 . The method of  claim 25  wherein said inhibition of expression of endogenous apoptosis-specific eIF-5A in said cell has an effect on the cell selected from the group consisting of suppressing apoptosis in said cell, reducing expression of p53 in said cell, reducing levels of a cytokine produced in said cell, reducing levels of a cytokine produced in said cell, increasing expression of Bcl-2 in said cell; reducing levels of myeloperoxidase produced in said cell, and reducing levels of active NFk beta in said cell, reducing levels of TLR4 in said cell, reducing levels of TNFR-1 in said cell and reducing levels of iNOS in said cell.  
     
     
         28 . The method of  claim 26  herein said inhibition of expression of endogenous apoptosis-specific eIF-5A in said cell has an effect on the cell selected from the group consisting of suppressing apoptosis in said cell, reducing expression of p53 in said cell, reducing levels of a cytokine produced in said cell, reducing levels of a cytokine produced in said cell, increasing expression of Bcl-2 in said cell; reducing levels of myeloperoxidase produced in said cell, and reducing levels of active NFk beta in said cell, reducing levels of TLR4 in said cell, reducing levels of TNFR-1 in said cell and reducing levels of iNOS in said cell.