Treatment of interferon regulatory factor 8 (IRF8) related diseases by inhibition of natural antisense transcript to IRF8
The present invention relates to antisense oligonucleotides that modulate the expression of and/or function of Interferon Regulatory Factor 8 (IRF8), in particular, by targeting natural antisense polynucleotides of Interferon Regulatory Factor 8 (IRF8). The invention also relates to the identification of these antisense oligonucleotides and their use in treating diseases and disorders associated with the expression of IRF8.
1. A method of testing an oligonucleotide for modulatory activity against an Interferon Regulatory Factor 8 (IRF8) polynucleotide comprising selecting a target IRF8 polynucleotide comprising SEQ ID NO: 1; identifying at least one natural antisense polynucleotide antisense to said target polynucleotide; identifying modified antisense oligonucleotides comprising oligonucleotides between 10 and 30 nucleotides in length which are complementary to the identified natural antisense polynucleotide; and, measuring the thermal melting point between binding of the modified antisense oligonucleotide and target natural antisense polynucleotide under stringent hybridization conditions; measuring the modulatory activity of the modified antisense oligonucleotide.
2. The method of claim 1 wherein the natural antisense polynucleotide has SEQ ID NO: 2.
3. The method of claim 1 , wherein the modified antisense oligonucleotide comprises one or more modifications selected from: at least one modified sugar moiety, at least one modified intemucleoside linkage, at least one modified nucleotide, and combinations thereof.
4. The method of claim 1 , wherein the one or more modifications comprise at least one modified sugar moiety selected from: a 2′-O-methoxyethyl modified sugar moiety, a 2′-methoxy modified sugar moiety, a 2′-O-alkyl modified sugar moiety, a bicyclic sugar moiety, and combinations thereof.
5. The method of claim 1 , wherein the one or more modifications comprise at least one modified intemucleoside linkage selected from: a phosphorothioate, 2′-O-methoxyethyl (MOE), 2′-fluoro, alkylphosphonate, phosphorodithioate, alkylphosphonothioate, phosphoramidate, carbamate, carbonate, phosphate triester, acetamidate, carboxymethyl ester, and combinations thereof.
6. The method of claim 1 , wherein the one or more modifications comprise at least one modified nucleotide selected from: a peptide nucleic acid (PNA), a locked nucleic acid (LNA), an arabino-nucleic acid (FANA), an analogue, a derivative, and combinations thereof.
7. The method of claim 1 , wherein the modified oligonucleotide comprises at least one oligonucleotide sequences set forth as SEQ ID NOS: 3 to 6.
8. The method of claim 3 , wherein said at least one modification comprises at least one phosphorothioate internucleotide linkage.
9. The oligonucleotide of claim 6 , wherein said oligonucleotide comprises a backbone of phosphorothioate internucleotide linkages.
10. The method of claim 1 , wherein the at least one modification comprises a plurality of modifications, wherein said modifications comprise modified sugar moieties selected from: a 2′-O-methoxyethyl modified sugar moiety, a 2′-methoxy modified sugar moiety, a 2′-O-alkyl modified sugar moiety, a bicyclic sugar moiety, and a combination thereof.
11. The method of claim 1 wherein the at least one modified oligonucleotide of 10 to 30 nucleotides in length upregulates the expression of the IRF8 gene.