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