Treatment of NANOG related diseases by inhibition of natural antisense transcript to NANOG
The present invention relates to antisense oligonucleotides that modulate the expression of and/or function of NANOG, in particular, by targeting natural antisense polynucleotides of NANOG. The invention also relates to the identification of these antisense oligonucleotides and their use in treating diseases and disorders associated with the expression of NANOG.
1. A pharmaceutical composition comprising a synthetic, modified oligonucleotide of 10 to 30 nucleotides in length comprising at least one modification wherein the at least one modification is selected from: at least one modified sugar moiety; at least one modified internucleotide linkage; at least one modified nucleotide, and combinations thereof; wherein said oligonucleotide is a single-stranded antisense compound which specifically hybridizes to a target region of a natural antisense polynucleotide of a NANOG gene having SEQ ID NO: 2 and is at least 90% complementary to said natural antisense polynucleotide and upregulates the function and/or expression of an NANOG gene in vivo or in vitro as compared to a normal control and a pharmaceutically acceptable excipient.
2. The composition according to claim 1 wherein said oligonucleotide is 10 to 30 nucleotides in length and has at least 95% sequence identity to the reverse complement of 10-30 nucleotides within a natural antisense transcript of the NANOG gene.
3. The composition of claim 1 , wherein the at least one modification comprises an internucleotide linkage selected from the group consisting of phosphorothioate, alkylphosphonate, phosphorodithioate, alkylphosphonothioate, phosphoramidate, carbamate, carbonate, phosphate triester, acetamidate, carboxymethyl ester, and combinations thereof.
4. The composition of claim 1 , wherein said oligonucleotide comprises at least one phosphorothioate internucleotide linkage.
5. The composition of claim 1 , wherein said oligonucleotide comprises a backbone of phosphorothioate internucleotide linkages.
6. The composition of claim 1 , wherein the oligonucleotide comprises at least one modified nucleotide, said modified nucleotide selected from: a peptide nucleic acid, a locked nucleic acid (LNA), analogue, derivative, and a combination thereof.
7. The composition of claim 1 , wherein the oligonucleotide comprises a plurality of modifications, wherein said modifications comprise modified nucleotides selected from: phosphorothioate, alkylphosphonate, phosphorodithioate, alkylphosphonothioate, phosphoramidate, carbamate, carbonate, phosphate triester, acetamidate, carboxymethyl ester, and a combination thereof.
8. The composition of claim 1 , wherein the oligonucleotide comprises a plurality of modifications, wherein said modifications comprise modified nucleotides selected from: peptide nucleic acids, locked nucleic acids (LNA), analogues, derivatives, and a combination thereof.
9. The composition of claim 1 , wherein the oligonucleotide comprises 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 a combination thereof.
10. The composition of claim 1 , wherein the oligonucleotide 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. An oligonucleotide which comprises the sequences set forth as SEQ ID NOS: 3 to 5.
12. A pharmaceutical composition comprising one or more oligonucleotides according to claim 11 and a pharmaceutically acceptable excipient.
13. The composition of claim 12 , wherein the oligonucleotides have at least about 90% sequence identity as compared to any one of the nucleotide sequences set forth as SEQ ID NOS: 3 to 5.
14. The composition of claim 12 , wherein the oligonucleotides comprise nucleotide sequences set forth as SEQ ID NOS: 3 to 5.
15. The composition of claim 14 , wherein the oligonucleotides set forth as SEQ ID NOS: 3 to 5 comprise one or more modifications or substitutions.
16. The composition of claim 15 , wherein the one or more modifications are selected from: phosphorothioate, methylphosphonate, peptide nucleic acid, locked nucleic acid (LNA) molecules, and combinations thereof.