Competitive modulation of micrornas

Abstract

The present invention provides compounds and methods for competitive modulation of microRNAs. Such copounds and methods have profound effects on cells.

1 - 135 . (canceled) 
     
     
         136 . A method of modulating miR-tone in a cell comprising contacting the cell with a broad competitive microRNA modulating compound, wherein the cell is in a human. 
     
     
         137 . The method of  claim 136 , wherein the modulating comprises a decrease in the activity and/or amount of a plurality of natural microRNAs. 
     
     
         138 . The method of  claim 136 , wherein the modulating comprises an increase in the activity and/or amount of a plurality of natural microRNAs. 
     
     
         139 . (canceled) 
     
     
         140 . The method of  claim 136 , wherein the human has a condition selected from among: cancer, inflammatory disease, metabolic disease, infectious disease, autoimmune disease, neurodegenerative disease and cardiovascular disease. 
     
     
         141 . (canceled) 
     
     
         142 . The method of  claim 137  wherein the natural microRNAs are selected from among mir-16, mir-17, mir-21, mir-24, mir-27a, mir-92, mir-202, mir-378, and mir-422a. 
     
     
         143 . (canceled) 
     
     
         144 . The method of  claim 136 , wherein the human has a condition selected from among: leukemia, lymphomas, multiple myeloma, ovarian neoplasms, colorectal cancer, pituitary neoplasms, adrenocortical carcinoma, prostatic neoplasms, Patau syndrome, non-small cell lung carcinoma, glioblastoma, breast neoplasms, myocardial infarction, Alzheimer, Parkinson, Hodgkin, medulloblastoma, multiple sclerosis, mesothelioma, obesity, periodontitis, pancreatic neoplasms, psoriasis, diabetes, eclampsia, Sezary syndrome, hypertension, fibrosis, Schizophrenia, myeolodysplastic syndrome, heptatocellular carcinoma, endometriosis, laryngeal neoplasms, systemic lupus erythematosus (SLE), rheumatoid arthritis, asthma, osteoarthritis, ulcerative colitis and atopic eczema. 
     
     
         145 . The method of claim  1 , wherein the method comprises performing an assay for determining whether a test compound is capable of competing with one or more natural microRNA for a microRNA-associated protein comprises:
 a. creating a first test sample comprising a test compound, a microRNA, and a microRNA-associated protein, wherein the test compound is at a first concentration;   b. creating a second test sample comprising the test compound, the microRNA, and the microRNA-associated protein, wherein the test compound is present at a second concentration;   c. determining the amount of test compound and/or the amount of microRNA bound to the microRNA-associated protein in each of the first and second test sample; and   d. calculating the relative binding affinity of the test compound and the microRNA for their binding for the microRNA-associated protein.   
     
     
         146 . The method of  claim 145 , wherein the assay for determining whether a test compound is capable of competing with one or more natural microRNA for a microRNA-associated protein comprises:
 a. creating a first test sample comprising a test compound, a microRNA, and a microRNA-associated protein, wherein the test compound is at a first concentration;   b. creating a second test sample comprising the test compound, the microRNA, and the microRNA-associated protein, wherein the test compound is present at a second concentration;   c. determining the activity of at least one microRNA in each of the first and second test sample; and;   d. calculating the degree to which the test sample alters activity of the at least one microRNA.   
     
     
         147 . The method of  claim 145 , wherein the microRNA-associated protein is provided at excess. 
     
     
         148 . The method of  claim 137 , wherein the natural microRNAs are selected from among mir-133, mir-331, mir-339, mir-532, and mir-615. 
     
     
         149 . The method of  claim 136 , wherein the broad competitive microRNA modulating compound is an oligonucleotide. 
     
     
         150 . The method of  claim 149 , wherein the broad competitive microRNA modulating compound is a single-stranded oligonucleotide 
     
     
         151 . The method of  claim 149 , wherein the broad competitive microRNA modulating compound is a double-stranded oligonucleotide. 
     
     
         152 . The method of  claim 136 , wherein the broad competitive microRNA modulating compound modulates the amount or activity of at least one object nucleic acid in the cells. 
     
     
         153 . The method of  claim 136 , wherein the broad competitive microRNA modulating compound modulates the amount or activity of at least one object protein in the cell. 
     
     
         154 . The method of  claim 153 , wherein the at least one object protein is a protease. 
     
     
         155 . The method of  claim 154 , wherein the protease is granzyme B. 
     
     
         156 . The method of  claim 154 , wherein the protease is granzyme M. 
     
     
         157 . The method of  claim 136 , wherein the broad competitive microRNA modulating compound ultimately modulates the amount or activity of α-tubulin. 
     
     
         158 . The method of  claim 149 , wherein the oligonucleotide comprises at least one modified nucleoside comprising a 2′-substituted sugar moiety.