Treatment of insulin gene (ins) related diseases by inhibition of natural antisense transcript to an insulin gene (ins)

Abstract

The present invention relates to antisense oligonucleotides that modulate the expression of and/or function of an Insulin gene (INS), in particular, by targeting natural antisense polynucleotides of an Insulin gene (INS). The invention also relates to the identification of these antisense oligonucleotides and their use in treating diseases and disorders associated with the expression of Insulin Gene (INS).

What is claimed is: 
     
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         14 . A method of modulating a function of and/or the expression of an Insulin gene (INS) in mammalian cells or tissues in vivo or in vitro comprising: contacting said cells or tissues with at least one short interfering RNA (siRNA) oligonucleotide 5 to 30 nucleotides in length, said at least one siRNA oligonucleotide being specific for an antisense polynucleotide of an Insulin gene (INS) polynucleotide, wherein said at least one siRNA oligonucleotide has at least 50% sequence identity to a complementary sequence of at least about five consecutive nucleic acids of the antisense and/or sense nucleic acid molecule of the Insulin gene (INS) polynucleotide; and, modulating a function of and/or the expression of an Insulin gene (INS) in mammalian cells or tissues in vivo or in vitro. 
     
     
         15 . The method of  claim 14 , wherein said oligonucleotide has at least 80% sequence identity to a sequence of at least about five consecutive nucleic acids that is complementary to the antisense and/or sense nucleic acid molecule of the Insulin gene (INS) polynucleotide. 
     
     
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         17 . 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 an antisense compound which specifically hybridizes to a natural antisense polynucleotide of an insulin (INS) gene and upregulates the function and/or expression of said Insulin gene (INS) in vivo or in vitro as compared to a normal control. 
     
     
         18 . The oligonucleotide of  claim 17 , 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. 
     
     
         19 . The oligonucleotide of  claim 17 , wherein said oligonucicotide comprises at least one phosphorothioate internucleotide linkage. 
     
     
         20 . The oligonucleotide of  claim 17 , wherein said oligonucleotide comprises a backbone of phosphorothioate internucleotide linkages. 
     
     
         21 . The oligonucleotide of  claim 17 , 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. 
     
     
         22 . The oligonucleotide of  claim 17 , 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. 
     
     
         23 . The oligonucleotide of  claim 17 , 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. 
     
     
         24 . The oligonucleotide of  claim 17 , 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. 
     
     
         25 . The oligonucleotide of  claim 17 , 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. 
     
     
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         29 . The oligonucleotide of  claim 17 , wherein the oligonucleotide comprises the sequences set forth as SEQ ID NOS: 4 to 11. 
     
     
         30 . A composition comprising one or more oligonucleotides according to  claim 17  and a pharmaceutically acceptable excipient. 
     
     
         31 . The composition of  claim 30 , wherein the oligonucleotides have at least but 40% sequence identity as compared to any one of the nucleotide sequences set forth as SEQ ID NOS: 4 to 11. 
     
     
         32 . The composition of  claim 30 , wherein the oligonucleotides comprise nucleotide sequences set forth as SEQ ID NOS: 4 to 11. 
     
     
         33 . The composition of  claim 32 , wherein the oligonucleotides set forth as SEQ ID NOS: 4 to 11 comprise one or more modifications or substitutions. 
     
     
         34 . The composition of  claim 33 , wherein the one or more modifications are selected from: phosphorothioate, methylphosphonate, peptide nucleic acid, locked nucleic acid (LNA) molecules, and combinations thereof. 
     
     
         35 . A method of preventing or treating a disease associated with at least one Insulin gene (INS) polynucleotide and/or at least one encoded product thereof, comprising: administering to a patient a therapeutically effective dose of at least one antisense oligonucleotide that binds to a natural antisense sequence of said at least one Insulin gene (INS) polynucleotide and modulates expression of said at least one Insulin gene (INS) polynucleotide; thereby preventing or treating the disease associated with the at least one Insulin gene (INS) polynucleotide and/or at least one encoded product thereof. 
     
     
         36 . The method of  claim 35 , wherein a disease associated with the at least one Insulin gene (INS) polynucleotide is selected from: a disease or disorder associated with abnormal function and/or expression of INS, diabetes or related disorders thereof (e.g., type I, type II diabetes, gestational diabetes, diabetic ketoacidosis, nonketotic hyperosmolar coma, hypoglycemia, diabetic coma, Neonatal diabetes mellitus, Permanent Neonatal diabetes mellitus, Insulin resistance, obesity, etc.), a metabolic disease or disorder (e.g., an insulin resistant non diabetic state, obesity, impaired glucose tolerance (IGT), Metabolic Syndrome, MODY syndrome, diabetic retinopathy, insulinoma, adult-onset diabetes, diabetic nephropathy, hyperglycemia, diabetic nephropathy, hypercholesterolemia, hyperinsulinism, dyslipidemia hyperlipidemia and an age-related metabolic disease etc.), a disease or disorder associated with impaired regulation of insulin level, neuropathy (e.g., sensory neuropathy, autonomic neuropathy, motor neuropathy, retinopathy), Polycystic Ovary Syndrome, cancer, inflammation, a neurological disease or disorder, a cardiac disease or disorder, an autoimmune disease, a prostatic neoplasm, hirsuitism and hypertension. 
     
     
         37 . A method of identifying and selecting at least one oligonucleotide for in vivo administration comprising: selecting a target polynucleotide associated with a disease state; identifying at least one oligonucleotide comprising at least five consecutive nucleotides which are complementary to the selected target polynucleotide or to a polynucleotide that is antisense to the selected target polynucleotide; measuring the thermal melting point of a hybrid of an antisense oligonucleotide and the target polynucleotide or the polynucleotide that is antisense to the selected target polynucleotide under stringent hybridization conditions; and selecting at least one oligonucleotide for in vivo administration based on the information obtained.