Mir-92 inhibitors and uses thereof

Abstract

The present invention provides oligonucleotide inhibitors of miR-92 and methods of using said inhibitors for inhibiting the function and/or activity of miR-92 in a subject in need thereof. The present invention also provides methods for evaluating or monitoring the efficacy of a therapeutic for promoting wound healing and selecting a subject for treatment with a therapeutic that modulates miR-92 function and/or activity.

1 . A method for promoting wound healing in a subject comprising administering an oligonucleotide comprising a sequence that is at least partially complementary to miR-92, wherein the administration of the oligonucleotide reduces function or activity of miR-92, thereby promoting wound healing. 
     
     
         2 . The method of  claim 1 , wherein the oligonucleotide comprises at least one locked nucleic acid (LNA) containing a 2′ to 4′ methylene bridge. 
     
     
         3 . The method of  claim 1  or  2 , wherein the oligonucleotide comprising a sequence that is at least partially complementary to miR-92 comprises a sequence of at least 16 nucleotides, wherein the sequence comprises no more than three contiguous LNAs, wherein from the 5′ end to the 3′ end, positions 1, 6, 10, 11, 13 and 16 of the sequence are LNAs. 
     
     
         4 . The method of  claim 3 , wherein from the 5′ end to the 3′ end, the sequence further comprises LNAs at positions 3, 9, and 14. 
     
     
         5 . The method of  claim 3 , wherein from the 5′ end to the 3′ end, the sequence further comprises LNAs at positions 3, 8, and 14. 
     
     
         6 . The method of  claim 3 , wherein from the 5′ end to the 3′ end, the sequence further comprises LNAs at positions 5, 8, and 15. 
     
     
         7 . The method of any one of  claims 4 - 6 , wherein from the 5′ end to the 3′ end, the sequence further comprises a deoxyribonucleic acid (DNA) nucleotide at the second nucleotide position. 
     
     
         8 . The method of  claim 7 , wherein the DNA nucleotide at the second nucleotide position contains a chemically modified nitrogenous base. 
     
     
         9 . The method of  claim 8 , wherein the chemically modified nitrogenous base is 5-methylcytosine. 
     
     
         10 . The method of any one of  claims 1 - 9 , wherein the oligonucleotide comprises at least one nucleotide that is 2′-deoxy, 2′ O-alkyl or 2′ halo modified. 
     
     
         11 . The method of any one of  claims 1 - 10 , wherein the oligonucleotide has a 5′ cap structure, 3′ cap structure, or 5′ and 3′ cap structure. 
     
     
         12 . The method of any one of  claims 1 - 11 , wherein the oligonucleotide comprises one or more phosphorothioate linkages. 
     
     
         13 . The oligonucleotide of  claim 12 , wherein the oligonucleotide is fully phosphorothioate-linked. 
     
     
         14 . The method of any one of  claims 1 - 13 , further comprising a pendent lipophilic group. 
     
     
         15 . The method of  claim 1 , wherein the oligonucleotide comprises a sequence selected from Table 1 or 2. 
     
     
         16 . The method of any one of  claims 1 - 15 , wherein the subject is human. 
     
     
         17 . The method of any one of  claims 1 - 16 , wherein the subject suffers from diabetes. 
     
     
         18 . The method of any one of  claims 1 - 17 , wherein the wound healing is for a chronic wound, diabetic foot ulcer, venous stasis leg ulcer or pressure sore. 
     
     
         19 . The method of any one of  claims 1 - 18 , wherein the administration of the oligonucleotide produces an increased rate of re-epithelialization, granulation, and/or neoangiogenesis during wound healing as compared to no treatment or treatment with an agent known to promote wound healing. 
     
     
         20 . The method of  claim 19 , wherein the agent known to promote wound healing is platelet derived growth factor (PDGF) or vascular endothelial growth factor (VEGF). 
     
     
         21 . An oligonucleotide comprising a sequence selected from Table 2. 
     
     
         22 . The oligonucleotide of  claim 21 , wherein at least one non-locked nucleotide of the oligonucleotide is 2′ deoxy, 2′ O-alkyl or 2′ halo modified. 
     
     
         23 . The oligonucleotide of  claim 21  or  22 , wherein at least one locked nucleic acid (LNA) of the oligonucleotide has a 2′ to 4′ methylene bridge. 
     
     
         24 . The oligonucleotide of any one of  claims 21 - 23 , wherein the oligonucleotide has a 5′ cap structure , 3′ cap structure, or 5′ and 3′ cap structure. 
     
     
         25 . The oligonucleotide of any one of  claims 21 - 24 , wherein the oligonucleotide comprises one or more phosphorothioate linkages. 
     
     
         26 . The oligonucleotide of  claim 25 , wherein the oligonucleotide is fully phosphorothioate-linked. 
     
     
         27 . The oligonucleotide of any one of  claims 21 - 26 , further comprising a pendent lipophilic group. 
     
     
         28 . A pharmaceutical composition comprising an effective amount of the oligonucleotide of any one of  claims 21 - 27 , or a pharmaceutically-acceptable salt thereof, and a pharmaceutically-acceptable carrier or diluent. 
     
     
         29 . The pharmaceutical composition of  claim 28 , wherein the pharmaceutically-acceptable carrier comprises a colloidal dispersion system, macromolecular complex, nanocapsule, microsphere, bead, oil-in-water emulsion, micelle, mixed micelle, or liposome. 
     
     
         30 . A method of reducing or inhibiting activity of miR-92 in a cell comprising contacting the cell with the oligonucleotide of any one of  claims 21 - 27 . 
     
     
         31 . The method of  claim 30 , wherein the cell is a mammalian cell. 
     
     
         32 . The method of  claim 31 , wherein the cell is a cardiac cell, muscle cell, fibrocyte, fibroblast, keratinocyte or endothelial cell. 
     
     
         33 . The method of any one of  claims 30 - 32 , wherein the cell is in vitro, in vivo or ex vivo. 
     
     
         34 . A method of promoting angiogenesis in a subject comprising administering to the subject the oligonucleotide of any one of  claims 21 - 27 . 
     
     
         35 . The method of  claim 34 , wherein the subject suffers from ischemia, myocardial infarction, chronic ischemic heart disease, peripheral or coronary artery occlusion, ischemic infarction, stroke, atherosclerosis, acute coronary syndrome, coronary artery disease, carotid artery disease, diabetes, chronic wound(s), peripheral vascular disease or peripheral artery disease. 
     
     
         36 . The method of  claim 34  or  35 , wherein the subject is a human. 
     
     
         37 . A method of treating diabetes, chronic wound(s), ischemia, myocardial infarction, chronic ischemic heart disease, peripheral or coronary artery occlusion, ischemic infarction, stroke, atherosclerosis, acute coronary syndrome, coronary artery disease, carotid artery disease, or peripheral artery disease comprising administering to the subject the oligonucleotide of any one of  claims 21 - 27 . 
     
     
         38 . The method of  claim 37 , wherein the subject is a human. 
     
     
         39 . A method for evaluating or monitoring the efficacy of a therapeutic for modulating angiogenesis in a subject receiving the therapeutic comprising:
 a) measuring the expression of one or more genes listed in Table 3 in a sample from the subject; and   b) comparing the expression of the one or more genes to a pre-determined reference level or level of the one or more genes in a control sample, wherein the comparison is indicative of the efficacy of the therapeutic.   
     
     
         40 . The method of  claim 39 , further comprising performing a walk time test on the subject, determining an ankle-bronchial index (ABI) for the subject, performing an arteriography or angiography on the subject, or performing a SPECT analysis on the subject. 
     
     
         41 . A method for evaluating or monitoring the efficacy of a therapeutic for modulating wound healing in a subject receiving the therapeutic comprising:
 a) measuring the expression of one or more genes listed in Table 3 in a sample from the subject; and   b) comparing the expression of the one or more genes to a pre-determined reference level or level of the one or more genes in a control sample, wherein the comparison is indicative of the efficacy of the therapeutic.   
     
     
         42 . The method of any one of  claims 39 - 41 , wherein the therapeutic modulates miR-92 function and/or activity. 
     
     
         43 . The method of  claim 42 , wherein the therapeutic is a miR-92 oligonucleotide inhibitor. 
     
     
         44 . The method of  claim 43 , wherein the miR-92 oligonucleotide inhibitor is selected from Tables 1 and 2. 
     
     
         45 . The method of any one of  claims 39 - 44 , wherein the subject suffers from ischemia, myocardial infarction, chronic ischemic heart disease, peripheral coronary artery occlusion, ischemic infarction, stroke, atherosclerosis, acute coronary syndrome, coronary artery disease, carotid artery disease, diabetes, chronic wound(s), peripheral vascular disease or peripheral artery disease. 
     
     
         46 . The method of any one of  claims 39 - 45 , wherein the subject is a human. 
     
     
         47 . A method for evaluating an agent's ability to promote angiogenesis or wound healing comprising:
 a) measuring the expression of one or more genes listed in Table 3 in a cell contacted with the agent; and   b) comparing the expression of the one or more genes to a pre-determined reference level or level of the one or more genes in a control sample, wherein the comparison is indicative of the agent's ability to promote angiogenesis or wound healing.   
     
     
         48 . The method of  claim 47 , further comprising determining miR-92 function and/or activity in the cell contacted with the agent. 
     
     
         49 . The method of  claim 47  or  48 , wherein the cell is a mammalian cell. 
     
     
         50 . The method of  claim 49 , wherein the cell is a cardiac cell, muscle cell, fibrocyte, fibroblast, keratinocyte or endothelial cell. 
     
     
         51 . The method of any one of  claims 47 - 50 , wherein the cell is in vitro, in vivo or ex vivo. 
     
     
         52 . A method for selecting a subject for treatment with a therapeutic that modulates miR-92 function and/or activity comprising:
 a) measuring the expression of one or more genes listed in Table 3 in a sample from the subject, wherein the subject is treated with the therapeutic; and   b) comparing the expression of the one or more genes to a pre-determined reference level or level of the one or more genes in a control sample, wherein the comparison is indicative of whether the subject should be selected for treatment with the therapeutic.   
     
     
         53 . The method of  claim 52 , further comprising further comprising performing a walk time test on the subject, determining an ankle-bronchial index (ABI) for the subject, performing an arteriography or angiography on the subject, or performing a SPECT analysis on the subject. 
     
     
         54 . The method of  claim 52 , further comprising determining miR-92 function and/or activity in the sample. 
     
     
         55 . The method of any one of  claim 52 - 54 , wherein the therapeutic is a miR-92 oligonucleotide inhibitor. 
     
     
         56 . The method of  claim 55 , wherein the miR-92 oligonucleotide inhibitor is selected from Tables 1 and 2. 
     
     
         57 . The method of any one of  claims 52 - 56 , wherein the subject suffers from ischemia, myocardial infarction, chronic ischemic heart disease, peripheral or coronary artery occlusion, ischemic infarction, stroke, atherosclerosis, acute coronary syndrome, coronary artery disease, carotid artery disease, diabetes, chronic wound(s), peripheral vascular disease or peripheral artery disease. 
     
     
         58 . The method of any one of  claims 52 - 57 , wherein the subject is a human.