Treatment of atonal homolog 1 (ATOH1) related diseases by inhibition of natural antisense transcript to ATOH1
The present invention relates to antisense oligonucleotides that modulate the expression of and/or function of Atonal homolog 1 (ATOH1), in particular, by targeting natural antisense polynucleotides of Atonal homolog 1 (ATOH1). The invention also relates to the identification of these antisense oligonucleotides and their use in treating diseases and disorders associated with the expression of ATOH1.
1. A method of identifying at least one modified oligonucleotide of 10 to 30 nucleotides in length which specifically hybridizes to a natural antisense transcript of a ATOH1 polynucleotide and modulates the expression of said ATOH1 polynucleotide comprising: identifying at least one oligonucleotide comprising 10-30 consecutive nucleotides which is at least 90% complementary to a natural antisense polynucleotide of the ATOH1 polynucleotide; measuring the thermal melting point of a hybrid of said antisense oligonucleotide and the natural antisense polynucleotide of said ATOH1 polynucleotide under stringent hybridization conditions; measuring the ATOH1 polynucleotide modulatory activity of the antisense oligonucleotide in vitro and identifying at least one antisense oligonucleotide which modulates the expression of said ATOH1 polynucleotide in vitro.
2. The method 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.
3. The method of claim 1 , wherein said oligonucleotide comprises at least one phosphorothioate internucleotide linkage.
4. The method of claim 1 , wherein said oligonucleotide comprises a backbone of phosphorothioate internucleotide linkages.
5. The method 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.
6. The method 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.
7. The method 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.
8. The method 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.
9. The method 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.
10. The method of claim 1 , wherein the oligonucleotide is of at least about 12 to 30 nucleotides in length and hybridizes to a natural antisense sense strand of an Atonal homolog 1 (ATOH1) polynucleotide wherein said oligonucleotide has 100% sequence complementarity to said natural antisense polynucleotide of the Atonal homolog 1 (ATOH1) polynucleotide.
11. The method of claim 1 , wherein the oligonucleotide specifically hybridizes to a natural antisense polynucleotide having SEQ ID NO: 2.
12. The method of claim 1 , wherein said oligonucleotide hybridizes to and modulates expression and/or function of at least one Atonal homolog 1 (ATOH1) polynucleotide in vivo or in vitro, as compared to a normal control.
13. The method of claim 1 , wherein the oligonucleotide comprises the sequences set forth as SEQ ID NOS: 3 and 4.