Polynucleotides encoding low density lipoprotein receptor

Abstract

The invention relates to compositions and methods for the preparation, manufacture and therapeutic use of polynucleotide molecules encoding low density lipoprotein receptor comprising at least one mutation (e.g., an LDLR signally enhancing mutation).

1 . A polynucleotide encoding a low density lipoprotein receptor (LDLR) wherein the LDLR comprises at least one mutation in a domain selected from the group consisting of EGF-A domain, the intracellular domain and both the EGF-domain and the intracellular domain, wherein the polynucleotide comprises;
 (a) a first region of linked nucleosides, said first region encoding a polypeptide of interest, said polypeptide of interest selected from the group consisting of SEQ ID NOs 37-55 and 724-729;   (b) a first flanking region located 5′ relative to said first region comprising at least one 5′ terminal cap;   (c) a second flanking region located 3′ relative to said first region comprising a 3′ tailing sequence of linked nucleosides; and   wherein said polynucleotide comprises at least one chemically modified nucleoside.   
     
     
         2 . The polynucleotide of  claim 1 , wherein the first region of linked nucleosides comprises at least an open reading frame of a nucleic acid sequence, wherein the nucleic acid sequence selected from the group consisting of SEQ ID NOs: 56-718 and 732-737. 
     
     
         3 . The polynucleotide of  claim 1 , wherein the polypeptide of interest comprises at least two mutations. 
     
     
         4 . The polynucleotide of  claim 3 , wherein the at least two mutations are located in the intracellular domain. 
     
     
         5 . The polynucleotide of  claim 4 , wherein the two mutations are selected from the group consisting of K816R, K830R and C839A. 
     
     
         6 . The polynucleotide of  claim 5 , wherein the two mutations are K830R and C839A. 
     
     
         7 . The polynucleotide of  claim 5 , wherein the polypeptide of interest comprises three mutations in the intracellular domain and the three mutations are K816R, K830R and C839A. 
     
     
         8 . The polynucleotide of  claim 3 , wherein the polypeptide of interest comprises one mutation in the EGF-A domain and at least two mutations in the intracellular domain. 
     
     
         9 . The polynucleotide of  claim 8 , wherein the one mutation in the EGF-A domain is selected from the group consisting of N316A and L339D and the at least two mutations in the intracellular domain are selected from the group consisting of K816R, K830R and C839A. 
     
     
         10 . The polynucleotide of  claim 9 , wherein the one mutation in the EGF-A domain is N316A and the intracellular domain comprises the mutations K830R and C839A. 
     
     
         11 . The polynucleotide of  claim 9 , wherein the one mutant in the EGF-A domain is N316A and the intracellular domain comprises the mutations K816R, K830R and C839A. 
     
     
         12 . The polynucleotide of  claim 9 , wherein the one mutant in the EGF-A domain is L339D and the intracellular domain comprises the mutations K830R and C839A. 
     
     
         13 . The polynucleotide of  claim 9 , wherein the one mutant in the EGF-A domain is L339D and the intracellular domain comprises the mutations K816R, K830R and C839A. 
     
     
         14 . The polynucleotide of  claim 1 , wherein the domain is the intracellular domain and the at least one mutation prevent IDOL from degrading LDLR. 
     
     
         15 . The polynucleotide of  claim 1 , wherein the at least one chemically modified nucleoside is selected from the modifications of Table 5. 
     
     
         16 . The polynucleotide of  claim 15 , wherein the modification of Table 5 is a uridine modification. 
     
     
         17 . The polynucleotide of  claim 15 , wherein the modification of Table 5 is a cytidine modification. 
     
     
         18 . The polynucleotide of  claim 15 , wherein the polynucleotide comprises two chemically modified nucleosides. 
     
     
         19 . The polynucleotide of  claim 18 , wherein the first chemically modified nucleoside is a uridine and the second chemically modified nucleoside is a cytosine. 
     
     
         20 . A method for modulating cholesterol levels in plasma of a subject comprising contacting said subject with a composition comprising any of the polynucleotides of  claims 1 - 19 . 
     
     
         21 . A method for increasing the level or amount of LDLR on the cell surface comprising contacting said subject with a composition comprising any of the polynucleotides of  claims 1 - 19 .