Methods and Compositions for Treating Dystroglycanopathy Disorders

Abstract

The invention relates to synthetic polynucleotides encoding fukutin related protein (FKRP). The invention further relates to nucleic acid constructs comprising the synthetic polynucleotides and methods of using these synthetic polynucleotides to treat dystroglycanopathy disorders.

What is claimed is: 
     
         1 . A polynucleotide comprising a synthetic nucleotide sequence which has at least 90% identity to SEQ ID NO:1 and encodes a human fukutin-related protein (FKRP). 
     
     
         2 . The polynucleotide of  claim 1 , wherein the synthetic nucleotide sequence has a GC content that is reduced by about 5% to about 10% relative to the GC content of SEQ ID NO:2. 
     
     
         3 . The polynucleotide of  claim 1 , wherein the polynucleotide sequence is operably linked to a promoter. 
     
     
         4 . The polynucleotide of  claim 3 , wherein the promoter is a creatine kinase (CK) promoter, or a chicken β-actin promoter (CB). 
     
     
         5 . The polynucleotide of  claim 4 , wherein the promoter comprises an enhancer sequence. 
     
     
         6 . The polynucleotide of  claim 5 , wherein the enhancer sequence comprises a CMV enhancer, a muscle creatine kinase enhancer, and/or a myosin light chain enhancer. 
     
     
         7 . A vector comprising the polynucleotide of  claim 1 . 
     
     
         8 . The vector of  claim 7 , wherein the vector is a viral vector. 
     
     
         9 . The vector of  claim 8 , wherein the vector is an adeno-associated virus (AAV) vector. 
     
     
         10 . The vector of  claim 9 , wherein the AAV vector is an AAV type 1, AAV type 2, AAV type 3a, AAV type 3B, AAV type 4, AAV type 5, AAV type 6, AAV type 7, AAV type 8, AAV type 9, AAV type 10, AAV type 11, avian AAV, bovine AAV, canine AAV, equine AAV, or ovine AAV. 
     
     
         11 . The vector of  claim 9 , wherein the AAV vector is an AAV9 vector. 
     
     
         12 . The vector of  claim 7 , further comprising a mir 122 binding element. 
     
     
         13 . A transformed cell comprising the polynucleotide of  claim 1  and/or a vector comprising said polynucleotide. 
     
     
         14 . A transgenic animal comprising the polynucleotide  claim 1 , a vector comprising said polynucleotide and/or a transformed cell comprising the polynucleotide or vector. 
     
     
         15 . A method of increasing glycosylation of alpha-dystroglycan (α-DG) in a subject in need thereof, comprising: delivering to said subject the polynucleotide of  claim 1  and/or a vector comprising said polynucleotide, wherein the synthetic polynucleotide is expressed in said subject, thereby producing human FKRP and increasing glycosylation of α-DG. 
     
     
         16 . A method of delivering a nucleic acid to a cell comprising delivering to the cell a therapeutically effective amount of polynucleotide of  claim 1  and/or a vector comprising said polynucleotide. 
     
     
         17 . A method of delivering a nucleic acid to a subject in need thereof, comprising delivering to the subject a therapeutically effective amount of the polynucleotide  claim 1 , a vector comprising said polynucleotide and/or a transformed cell comprising the polynucleotide or vector. 
     
     
         18 . A method of treating a dystroglycanopathy in a subject in need thereof, comprising delivering to the subject a therapeutically effective amount of the polynucleotide  claim 1 , a vector comprising said polynucleotide and/or a transformed cell comprising the polynucleotide or vector, thereby treating dystroglycanopathy in the subject. 
     
     
         19 . The method of  claim 17 , wherein the dystroglycanopathy comprises a mutation in the nucleic acid encoding FKRP and/or a deficiency in glycosylation of alpha-dystroglycan (α-DG), or any combination thereof. 
     
     
         20 . The method of  claim 18 , wherein the dystroglycanopathy is limb girdle muscular dystrophy 2I, congenital muscular dystrophy, Walker-Warburg syndrome, muscle-eye-brain disease, or any combination thereof.