Methods of treatment with mini-dystrophin nucleic acid sequences

Abstract

The present invention relates to compositions and methods for expressing mini-dystrophin peptides. In particular, the present invention provides compositions comprising nucleic acid sequences that are shorter than wild-type dystrophin cDNA and that express mini-dystrophin peptides that function in a similar manner as wild-type dystrophin proteins. The present invention also provides compositions comprising mini-dystrophin peptides, and methods for expressing mini-dystrophin peptides in target cells.

1 - 34 . (cancelled).  
     
     
         35 . A method comprising; 
 a) providing; 
 i) a vector comprising nucleic acid which is less than 5.0 kb in length and which encodes a mini-dystrophin peptide, wherein said mini-dystrophin peptide comprises a spectrin-like repeat domain comprising 4 dystrophin spectrin-like repeats, and wherein said mini-dystrophin peptide is capable of increasing a diaphragm specific force value in a DMD animal model by at least 20% of the wild type value; and  
 ii) a subject comprising target cells; and  
   b) contacting said vector with said subject under conditions such that said mini-dystrophin peptide is expressed in said target cells.    
     
     
         36 . The method of  claim 35 , wherein said vector comprises an adeno-associated vector.  
     
     
         37 . The method of  claim 35 , wherein said contacting is done by means of injecting said vector into said subject.  
     
     
         38 . The method of  claim 35 , wherein said mini-dystrophin peptide is capable of increasing a diaphragm specific force value in a DMD animal model by at least 30% of the wild type value.  
     
     
         39 . The method of  claim 35 , wherein said dystrophin spectrin-like repeats are human dystrophin spectrin-like repeats.  
     
     
         40 . The method of  claim 35 , wherein said nucleic acid comprises an actin-binding domain encoding sequence.  
     
     
         41 . The method of  claim 40 , wherein said actin binding domain comprises at least a portion of SEQ ID NO:6.  
     
     
         42 . The method of  claim 35 , wherein said nucleic acid comprises a β-dystroglycan binding domain.  
     
     
         43 . The method of  claim 42 , wherein said β-dystroglycan binding domain comprises at least a portion of a dystrophin hinge 4 encoding sequence, and at least a portion of a dystrophin cysteine-rich domain encoding sequence.  
     
     
         44 . The method of  claim 35 , wherein said spectrin-like repeat encoding sequences are selected from the group consisting of SEQ ID NOS:8-10, 12-27, and 29-33.  
     
     
         45 . The method of  claim 35 , wherein said nucleic acid contains less than 75% of a wild type dystrophin 5′ untranslated region.  
     
     
         46 . The method of  claim 35 , wherein said mini-dystrophin peptide further comprises a substantially deleted dystrophin C-terminal domain.  
     
     
         47 . The method of  claim 35 , wherein said nucleic acid contains less than 50% of a dystrophin 3′ untranslated region.  
     
     
         48 . The method of  claim 35 , wherein said mini-dystrophin peptide further comprises dystrophin hinge region  1  and dystrophin hinge region  4 .  
     
     
         49 . The method of  claim 35 , wherein said mini-dystrophin peptide further comprises dystrophin hinge region  2  or dystrophin hinge region  3 .  
     
     
         50 . The method of  claim 35 , wherein said nucleic acid comprises a promoter.  
     
     
         51 . The method of  claim 50 , wherein said promoter is an MCK promoter.  
     
     
         52 . The method of  claim 35 , wherein said 4 dystrophin spectrin-like repeats are selected from the group consisting of: dystrophin spectrin-like repeat number 1, dystrophin spectrin-like repeat number 2, dystrophin spectrin-like repeat number 3, dystrophin spectrin-like repeat number 22, dystrophin spectrin-like repeat number 23, and dystrophin spectrin like repeat number 24.