Compositions and methods of altering cholesterol levels

Abstract

The present invention relates to compositions, methods and kits using polynucleotides, primary transcripts and mmRNA molecules.

What is claimed is: 
     
         1 . A composition comprising;
 (a) a synthetic polynucleotide encoding PCSK9 negative LDLR, and   (b) a synthetic polynucleotide encoding CYP7A1, in an acceptable diluent or carrier.   
     
     
         2 . The composition of  claim 1 , further comprising;
 (c) a statin selected from the group consisting of atorvastatin, cerivastatin, fluvastatin, lovastatin, mevastatin, pitavastatin, pravastatin, simvastatin, and combinations thereof.   
     
     
         3 . The composition of  claim 1 , wherein the synthetic polynucleotide comprises:
 (a) a first region of linked nucleosides, said first region encoding at least one cholesterol regulating polypeptide;   (b) a first flanking region located at the 5′ terminus of said first region comprising;
 (i) a sequence of linked nucleosides selected from the group consisting of the native 5′ untranslated region (UTR), SEQ ID NO: 1 and functional variants thereof; 
   (c) a second flanking region located at the 3′ terminus of said first region comprising;
 (i′) a sequence of linked nucleosides selected from the group consisting of the native 3′ UTR, any of the forgoing comprising one or more microRNA or microRNA binding site or microRNA seeds and functional variants or combinations thereof; and 
 (ii′) a 3′ tailing sequence of linked nucleosides; 
 wherein the first region of linked nucleosides comprises at least a first modified nucleoside. 
   
     
     
         4 . The composition of  claim 3  wherein the second flanking region encodes at least one miR binding site. 
     
     
         5 . The composition of  claim 4 , wherein said miR binding site is selected from the group consisting of miR-122a and miR-422a. 
     
     
         6 . The composition of any of  claims 1  wherein the synthetic polynucleotide comprises at least one chemical modification. 
     
     
         7 . A method of treating a disease or disorder in a subject in need thereof comprising administering to said subject the composition of any of  claims 1 . 
     
     
         8 . The method of  claim 7  further comprising measuring cholesterol levels in plasma of said subject. 
     
     
         9 . The method of  claim 7 , further comprising contacting said liver cell with a statin. 
     
     
         10 . The method of  claim 7  wherein the subject is a human. 
     
     
         11 . The method of  claim 10 , wherein the human has a polymorphism in CYP7A1. 
     
     
         12 . The method of  claim 7  wherein the disease or disorder is selected from the group consisting of fatty liver disease, hepatocellular carcinoma, NASH, steatosis, familial hypercholesterolemia (FH), hypercholesterolemia, and aberrant lipoprotein profile. 
     
     
         13 . The method of  claim 7  wherein the disease or disorder is familiar hypercholesterolemia (FH). 
     
     
         14 . The method of  claim 7  wherein the composition further comprises a pharmaceutically acceptable excipient. 
     
     
         15 . The method of  claim 7 , where the composition comprises a lipid and wherein said lipid is selected from DLin-DMA, DLin-K-DMA, DLin-KC2-DMA, 98N12-5, C12-200, DLin-MC3-DMA, reLNP, PLGA, PEG, PEG-DMA and PEGylated lipids and mixtures thereof. 
     
     
         16 . The method of  claim 7  wherein the synthetic polynucleotide is administered at a total daily dose of between 1 ug and 150 ug. 
     
     
         17 . A method of modulating cholesterol levels in plasma of a subject comprising contacting said subject with the composition of  claim 1 . 
     
     
         18 . The method of  claim 18 , wherein modulation is lowered levels.