Treatment of sirtuin (sirt) related diseases by inhibition of natural antisense transcript to a sirtuin (sirt)

Abstract

The present invention relates to antisense oligonucleotides that modulate the expression of and/or function of a Sirtuin (SIRT), in particular, by targeting natural antisense polynucleotides of a Sirtuin (SIRT). The invention also relates to the identification of these antisense oligonucleotides and their use in treating diseases and disorders associated with the expression of Sirtuins (SIRT)s.

1 . A method of a function of and/or the expression of a Sirtuin (SIRT) polynucleotide in patient cells or tissues in vivo or in vitro comprising: contacting said cells or tissues with at least one antisense oligonucleotide 5 to 30 nucleotides in length wherein said at least one oligonucleotide has at least 50% sequence identity to a reverse complement of a polynucleotide comprising 5 to 30 consecutive nucleotides within nucleotides 1 to 1028 of SEQ ID NO: 5 or nucleotides 1 to 429 of SEQ ID NO: 6, or nucleotides 1 to 156 of SEQ ID NO: 7 or nucleotides 1 to 593 of SEQ ID NO:8, 1 to 373 of SEQ ID NO: 9, 1 to 1713 of SEQ ID NO: 10, 1 to 660 of SEQ ID NO:11, 1 to 589 of SEQ ID NO: 12, 1 to 428 of SEQ ID NO: 13 and 1 to 4041 of SEQ ID NO: 14; thereby modulating a function of and/or the expression of the Sirtuin (SIRT) polynucleotide in patient cells or tissues in vivo or in vitro. 
     
     
         2 . A method of modulating a function of and/or the expression of a Sirtuin (SIRT) polynucleotide in patient cells or tissues in vivo or in vitro comprising: contacting said cells or tissues with at least one antisense oligonucleotide 5 to 30 nucleotides in length wherein said at least one oligonucleotide has at least 50% sequence identity to a reverse complement of a natural antisense of a Sirtuin (SIRT) polynucleotide; thereby modulating a function of and/or the expression of the Sirtuin (SIRT) polynucleotide in patient cells or tissues in vivo or in vitro. 
     
     
         3 . A method of modulating a function of and/or the expression of a Sirtuin (SIRT) polynucleotide in patient cells or tissues in vivo or in vitro comprising: contacting said cells or tissues with at least one antisense oligonucleotide 5 to 30 nucleotides in length wherein said oligonucleotide has at least 50% sequence identity to an antisense oligonucleotide to the Sirtuin (SIRT) polynucleotide; thereby modulating a function of and/or the expression of the Sirtuin (SIRT) polynucleotide in patient cells or tissues in vivo or in vitro. 
     
     
         4 . A method of modulating a function of and/or the expression of a Sirtuin (SIRT) polynucleotide in patient cells or tissues in vivo or in vitro comprising: contacting said cells or tissues with at least one antisense oligonucleotide that targets a region of a natural antisense oligonucleotide of the Sirtuin (SIRT) polynucleotide; thereby modulating a function of and/or the expression of the Sirtuin (SIRT) polynucleotide in patient cells or tissues in vivo or in vitro. 
     
     
         5 . The method of  claim 4 , wherein a function of and/or the expression of the Sirtuin (SIRT) is increased in vivo or in vitro with respect to a control. 
     
     
         6 . The method of  claim 4 , wherein the at least one antisense oligonucleotide targets a natural antisense sequence of a Sirtuin (SIRT) polynucleotide. 
     
     
         7 . The method of  claim 4 , wherein the at least one antisense oligonucleotide targets a nucleic acid sequence comprising coding and/or non-coding nucleic acid sequences of a Sirtuin (SIRT) polynucleotide. 
     
     
         8 . The method of  claim 4 , wherein the at least one antisense oligonucleotide targets overlapping and/or non-overlapping sequences of a Sirtuin (SIRT) polynucleotide. 
     
     
         9 . The method of  claim 4 , wherein the at least one antisense oligonucleotide comprises one or more modifications selected from: at least one modified sugar moiety, at least one modified internucleoside linkage, at least one modified nucleotide, and combinations thereof. 
     
     
         10 . The method of  claim 9 , 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. 
     
     
         11 . The method of  claim 9 , wherein the one or more modifications comprise at least one modified internucleoside linkage selected from: a phosphorothioate, 2′-Omethoxyethyl (MOE), 2′-fluoro, alkylphosphonate, phosphorodithioate, alkylphosphonothioate, phosphoramidate, carbamate, carbonate, phosphate triester, acetamidate, carboxymethyl ester, and combinations thereof. 
     
     
         12 . The method of  claim 9 , 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. 
     
     
         13 . The method of  claim 1 , wherein the at least one oligonucleotide comprises at least one oligonucleotide sequences set forth as SEQ ID NOS: 15 to 94. 
     
     
         14 . A method of modulating a function of and/or the expression of a Sirtuin (SIRT) 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 a Sirtuin (SIRT) 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 Sirtuin (SIRT) polynucleotide; and, modulating a function of and/or the expression of a Sirtuin (SIRT) 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 Sirtuin (SIRT) polynucleotide. 
     
     
         16 . A method of modulating a function of and/or the expression of a Sirtuin (SIRT) in mammalian cells or tissues in vivo or in vitro comprising: contacting said cells or tissues with at least one antisense oligonucleotide of about 5 to 30 nucleotides in length specific for noncoding and/or coding sequences of a sense and/or natural antisense strand of a Sirtuin (SIRT) polynucleotide wherein said at least one antisense oligonucleotide has at least 50% sequence identity to at least one nucleic acid sequence set forth as SEQ ID NOS: 1 to 14; and, modulating the function and/or expression of the Sirtuin (SIRT) in mammalian cells or tissues in vivo or in vitro. 
     
     
         17 . A synthetic, modified oligonucleotide 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 hybridizes to and modulates the function and/or expression of a Sirtuin (SIRT) 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 oligonucleotide 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. 
     
     
         26 . The oligonucleotide of  claim 17 , wherein the oligonucleotide is of at least about 5 to 30 nucleotides in length and hybridizes to an antisense and/or sense strand of a Sirtuin (SIRT) polynucleotide wherein said oligonucleotide has at least about 20% sequence identity to a complementary sequence of at least about five consecutive nucleic acids of the antisense and/or sense coding and/or noncoding nucleic acid sequences of the Sirtuin (SIRT) polynucleotide. 
     
     
         27 . The oligonucleotide of  claim 17 , wherein the oligonucleotide has at least about 80% sequence identity to a complementary sequence of at least about five consecutive nucleic acids of the antisense and/or sense coding and/or noncoding nucleic acid sequence of the Sirtuin (SIRT) polynucleotide. 
     
     
         28 . The oligonucleotide of  claim 17 , wherein said oligonucleotide hybridizes to and modulates expression and/or function of at least one Sirtuin (SIRT) polynucleotide in vivo or in vitro, as compared to a normal control. 
     
     
         29 . The oligonucleotide of  claim 17 , wherein the oligonucleotide comprises the sequences set forth as SEQ ID NOS: 15 to 94. 
     
     
         30 . A composition comprising one or more oligonucleotides specific for one or more Sirtuin (SIRT) polynucleotides, said polynucleotides comprising antisense sequences, complementary sequences, alleles, homologs, isoforms, variants, derivatives, mutants, fragments, or combinations thereof. 
     
     
         31 . The composition of  claim 30 , wherein the oligonucleotides have at least about 40% sequence identity as compared to any one of the nucleotide sequences set forth as SEQ ID NOS: 15 to 94. 
     
     
         32 . The composition of  claim 30 , wherein the oligonucleotides comprise nucleotide sequences set forth as SEQ NOS: 15 to 94. 
     
     
         33 . The composition of  claim 32 , wherein the oligonucleotides set forth as SEQ ID NOS: 15 to 94 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 Sirtuin (SIRT) 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 Sirtuin (SIRT) polynucleotide and modulates expression of said at least one Sirtuin (SIRT) polynucleotide; thereby preventing or treating the disease associated with the at least one Sirtuin (SIRT) polynucleotide and/or at least one encoded product thereof. 
     
     
         36 . The method of  claim 35 , wherein a disease associated with the at least one Sirtuin (SIR) polynucleotide is selected from cancer (e.g., breast cancer, colorectal cancer, CCL, CML, prostate cancer), a neurodegenerative disease or disorder (e.g., Alzheimer's Disease (AD), Huntington's disease, Parkinson's disease, Amyotrophic Lateral Sclerosis (ALS), Multiple Sclerosis, and disorders caused by polyglutamine agwegation); skeletal muscle disease (e.g., Duchene muscular dystrophy, skeletal muscle atrophy, Becker's dystrophy, or myotonic dystrophy); a metabolic disease or disorder (e.g., insulin resistance, diabetes, type 2 diabetes, obesity, impaired glucose tolerance, metabolic syndrome, adult-onset diabetes, diabetic nephropathy, hyperglycemia, diabetic nephropathy, Hypercholesterolemia, 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), a disease or disorder associated with a ketogenic condition, a disease or disorder associated with impaired energy homeostasis, a disease or disorder associated with impaired Acetyl-CoA synthetase 2 activity, a disease or disorder associated with metabolic homeostasis, a lipid metabolism disease or disorder, a disease or disorder associated with impaired thermogenesis, a disease or disorder associated with mitochondrial dysfunction, neuropathy (e.g., sensory neuropathy, autonomic neuropathy, motor neuropathy, retinopathy), a liver disease (e.g., due to alcohol abuse or hepatitis, fatty liver disease etc.); age-related macular degeneration, bone disease (e.g., osteoporosis), a blood disease (e.g., a leukemia); liver disease (e.g., due to alcohol abuse or hepatitis); obesity; bone resorption, age-related macular degeneration, AIDS related dementia, ALS, Bell's Palsy, atherosclerosis, a cardiac disease (e.g., cardiac dysrhymias, chronic congestive heart failure, ischemic stroke, coronary artery disease and cardiomyopathy), chronically degenerative disease (e.g., cardiac muscle disease), chronic renal failure, type 2 diabetes, ulceration, cataract, presbiopia, glomerulonephritis, Guillan-Barre syndrome, hemorrhagic stroke, rheumatoid arthritis, inflammatory bowel disease, SLE, Crohn's disease, osteoarthritis, osteoporosis, Chronic Obstructive Pulmonary Disease (COPD), pneumonia, skin aging, urinary incontinence, a disease or disorder associated with mitochondrial dysfunction (e.g., mitochondrial myopathy, encephalopathy, Leber's disease, Leigh encephalopathia, Pearson's disease, lactic acidosis, ‘mitochondrial encephalopathy; lactic acidosis and stroke like symptoms’ (MELAS) etc.) and a disease or disorder associated with neuronal cell death, degenerative syndrome, aging, a disease or disorder associated with telomere dysfunction, a disease or disorder associated with impaired chromatin regulation, a disease or disorder associated with premature cellular senescence, a disease or disorder associated with impared SIRT6 mediated DNA repair and a condition characterized by unwanted cell loss. 
     
     
         37 . A method of preventing or treating a skin condition associated with at least one Sirtuin (SIRT) polynucleotide and/or at least one encoded product thereof, comprising: administering to a patient having a skin condition or at risk of developing a skin condition a therapeutically effective dose of at least one antisense oligonucleotide that binds to a natural antisense sequence of said at least one Sirtuin (SIRT) polynucleotide and modulates expression of said at least one Sirtuin (SIRT) polynucleotide; thereby preventing or treating the disease skin condition associated with the at least one Sirtuin (SIRT) polynucleotide and/or at least one encoded product thereof. 
     
     
         38 . The method of  claim 38 , wherein the skin condition is caused by caused by inflammation, light damage or aging. 
     
     
         39 . The method of  claim 39 , wherein the skin condition is the development of wrinkles, contact dermatitis, ample dermatitis, actinic keratosis, keratinization disorders, an epidermolysis bullosa disease, exfoliative dermatitis, seborrheic dermatitis, an erythema, discoid lupus erythematosus, dermatomyositis, skin cancer, or an effect of natural aging.