Identification of a novel repressor on IFN-lambda promoter and siRNA against ZEB1 and BLIMP-1 to increase IFN-lambda gene activity
The present invention is directed to the identification of a novel repressor located between ˜1.2 kb to ˜1.6 kb from the translation start site of the IFN-λ1 promoter. The present invention provides a method of using siRNAs against ZEB1 (binds to the repressor region) and BLIMP-1 (binds outside the repressor region) and increases the promoter activity of IFN-λ1 (i.e., increases the production of IFN-λ1 protein). siRNAs against ZEB1 mRNA or BLIMP-1 mRNA increase IFN-λ1 gene activity. There is provided a therapeutic application of siRNAs against ZEB1 and BLIMP-1 mRNAs in treating a mammal (including a human) by increasing the production of IFN-λ1 protein that promotes an anti-viral response as well as treats asthma diseases.
1. A method of increasing IFN-λ1 gene activity in a mammalian cell, comprising the steps of:
(a) providing a mammalian cell in need of increasing IFN-λ1 gene activity, said mammalian cell is viral-stimulated; and
(b) exposing said mammalian cell to a siRNA oligonucleotide targeted against ZEB1 transcriptional factor mRNA, thereby increasing IFN-λ1 gene activity in said mammalian cell, as indicated by either an increase in IFN-λ1 mRNA or IFN-λ1 protein.
2. The method of claim 1 , wherein said siRNA oligonucleotide is at least one siRNA oligonucleotide selected from the group consisting of SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, and SEQ ID NO: 15, and said ZEB1 mRNA has a nucleotide sequence set forth in Accession No: NM — 030751 or Accession No: NM — 001128128.
3. The method of claim 1 , wherein said increased IFN-λ1 gene activity is indicated by an increase in IFN-λ1 mRNA.
4. The method of claim 3 , wherein said IFN-λ1 mRNA is measured by qPCR.
5. The method of claim 1 , wherein said mammalian cell is a human airway epithelial cell.
6. A method of treating a human subject inflicted with an asthmatic disease, comprising the step of administering a therapeutically effective amount of a siRNA oligonucleotide to said human subject, said siRNA oligonucleotide is targeted against ZEB1 mRNA, and induces the production of an IFN-λ1 protein having an amino acid sequence set forth in GenBank Accession No. NP — 742152.
7. The method of claim 6 wherein said siRNA oligonucleotide is at least one oligonucleotide selected from the group consisting of SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, and SEQ ID NO: 15, and said ZEB1 mRNA has a nucleotide sequence set forth in GenBank Accession No: NM — 030751 or GenBank Accession No: NM — 001128128.
8. The method of claim 1 , wherein said increased IFN-λ1 gene activity is indicated by an increase in IFN-λ1 protein.
9. The method of claim 8 , wherein said IFN-λ1 protein is measured by an ELISA.