Variant AAV and Compositions, Methods and Uses for Gene Transfer to Cells, Organs and Tissues

Abstract

The invention relates to adeno-associated virus (AAV) serotype AAV-Rh74 and related AAV vectors, and AAV-Rh74 and related AAV vector mediated gene transfer methods and uses. In particular, AAV-Rh74 and related AAV vectors target polynucleotides to cells, tissues or organs for expression (transcription) of genes encoding therapeutic proteins and peptides, and polynucleotides that function as or are transcribed into inhibitory nucleic acid sequences.

1 .- 6 . (canceled) 
     
     
         7 . A recombinant AAV particle, comprising the AAV capsid sequence of SEQ ID NO:5, wherein the genome of said recombinant AAV particle comprises an AAV inverted terminal repeat and a heterologous polynucleotide sequence that encodes an enzyme involved in a lysosomal storage disease, said polynucleotide operably linked to an expression control element. 
     
     
         8 .- 10 . (canceled) 
     
     
         11 . A method for delivering or transferring a heterologous polynucleotide sequence into a mammal, comprising administering a recombinant AAV particle comprising a recombinant vector genome comprising a heterologous polynucleotide sequence to said mammal, wherein the heterologous polynucleotide sequence encodes an enzyme involved in a lysosomal storage disease, thereby delivering or transferring the heterologous polynucleotide sequence into the mammal, wherein the recombinant AAV particle comprises the AAV capsid sequence of  claim 7 . 
     
     
         12 . (canceled) 
     
     
         13 . The method of  claim 11 , wherein the heterologous polynucleotide sequence is operably linked to an expression control element conferring transcription of said heterologous polynucleotide sequence. 
     
     
         14 . A method of treating a mammal deficient in an enzyme involved in a lysosomal storage disease, comprising: (a) providing a recombinant AAV particle comprising the AAV capsid sequence of SEQ ID NO:5 wherein the vector genome comprises a heterologous polynucleotide sequence encoding an enzyme that can correct for the deficient enzyme expression or function, wherein the heterologous polynucleotide sequence is operably linked to an expression control element conferring transcription of said heterologous polynucleotide sequence; and (b) administering an amount of the recombinant AAV particle to the mammal wherein said enzyme is expressed in the mammal 
     
     
         15 .- 28 . (canceled) 
     
     
         29 . The method of  claim 11  or  14 , wherein said enzyme involved in a lysosomal storage disease is expressed at levels having a therapeutic effect on the mammal 
     
     
         30 . The method of  claim 11  or  14 , wherein said adeno-associated virus (AAV) vector has increased tropism for hepatocytes compared to Rh74, AAV2 or AAV8 serotypes. 
     
     
         31 . The method of  claim 11  or  14 , wherein said enzyme involved in a lysosomal storage disease is expressed in a cell, tissue or organ of said mammal. 
     
     
         32 . The method of  claim 31 , wherein the cell comprises a secretory cell. 
     
     
         33 . The method of  claim 31 , wherein the cell comprises an endocrine cell. 
     
     
         34 . The method of  claim 31 , wherein the cell comprises hepatocyte, an epithelial cell, or a totipotent, pluripotent or multipotent stem cell. 
     
     
         35 . The method of  claim 31 , wherein the tissue or organ of said mammal comprises liver. 
     
     
         36 . The method of  claim 11  or  14 , wherein the mammal produces an insufficient amount of the enzyme involved in a lysosomal storage disease or a defective or aberrant enzyme involved in a lysosomal storage disease. 
     
     
         37 .- 43 . (canceled) 
     
     
         44 . The method of  claim 11  or  14 , wherein the expression control element comprises a constitutive or regulatable control element. 
     
     
         45 . The method of  claim 11  or  14 , wherein the expression control element comprises a tissue-specific expression control element or promoter. 
     
     
         46 . The method of  claim 11  or  14 , wherein the AAV vector is delivered intravenously, intraarterially, intramuscularly, subcutaneously, orally, by intubation, via catheter, dermally, intra-cranially, via inhalation, intra-cavity, or mucosally. 
     
     
         47 . The method of  claim 11  or  14 , wherein the mammal is human. 
     
     
         48 . The method of  claim 11  or  14 , wherein the mammal is sero-positive for an AAV serotype other than AAV-Rh74. 
     
     
         49 . The method of  claim 11  or  14 , wherein the mammal is sero-positive for an AAV serotype AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10 or AAV11. 
     
     
         50 . The method of  claim 11  or  14 , wherein the mammal is sero-negative or sero-positive for AAV-Rh74. 
     
     
         51 . The method of  claim 11  or  14 , further comprising administering empty capsid AAV. 
     
     
         52 . The method of  claim 11  or  14 , further comprising administering empty capsid AAV-Rh74 or empty capsid comprising the AAV capsid sequence of SEQ ID NO:5. 
     
     
         53 . (canceled) 
     
     
         54 . The method of  claim 11  or  14 , wherein the AAV particle comprises a pharmaceutical composition. 
     
     
         55 . The method of  claim 54 , wherein the pharmaceutical composition comprises empty capsid AAV. 
     
     
         56 . The method of  claim 54 , wherein the pharmaceutical composition comprises empty capsid AAV-Rh74 or empty capsid comprising the AAV capsid sequence of SEQ ID NO:5. 
     
     
         57 . The recombinant AAV particle of  claim 7 , wherein said expression control element comprises a liver-specific enhancer and promoter. 
     
     
         58 . The recombinant AAV particle of  claim 7 , wherein said heterologous polynucleotide sequence is linked to a poly-A sequence. 
     
     
         59 . The recombinant AAV particle of  claim 7 , wherein said AAV inverted terminal repeat is from AAV2. 
     
     
         60 . The recombinant AAV particle of  claim 7 , wherein said expression control element is an ApoE-hAAT enhancer-promoter. 
     
     
         61 . The recombinant AAV particle of  claim 7 , wherein said heterologous polynucleotide sequence is flanked by 5′ and 3′ untranslated regions. 
     
     
         62 . The method of  claim 11 , wherein the lysosomal storage disease is Pompe's disease. 
     
     
         63 . The method of  claim 14 , wherein the mammal has Pompe's disease. 
     
     
         64 . The method of  claim 45 , wherein said tissue-specific expression control element or promoter is specific for liver. 
     
     
         65 . A method for producing the recombinant AAV particle of  claim 7 , comprising culturing a helper cell comprising a recombinant plasmid comprising the genome of the recombinant AAV particle of  claim 7 . 
     
     
         66 . A recombinant AAV (rAAV) particle comprising a VP1 capsid protein having the amino acid sequence of SEQ ID NO:5, wherein the genome of said rAAV particle comprises, in 5′ to 3′ order:
 (a) a first AAV inverted terminal repeat, 
 (b) a liver-specific enhancer and promoter, 
 (c) a polynucleotide sequence that encodes an enzyme involved in a lysosomal storage disease operably linked to the liver-specific enhancer and promoter, 
 (d) a poly-A sequence; and 
 (e) a second AAV inverted terminal repeat. 
 
     
     
         67 . The rAAV particle of  claim 66 , wherein said first and second AAV inverted terminal repeats are from AAV2. 
     
     
         68 . The rAAV particle of  claim 66 , wherein said liver-specific enhancer and promoter is an ApoE-hAAT enhancer-promoter. 
     
     
         69 . The rAAV particle of  claim 66 , wherein said polynucleotide sequence is flanked by 5′ and 3′ untranslated regions. 
     
     
         70 . A pharmaceutical composition comprising the rAAV particle of  claim 7  or  66  and a pharmaceutically acceptable carrier. 
     
     
         71 . A method of treating a subject in need of treatment for Pompe's disease comprising administering to said subject a therapeutically effective amount of the pharmaceutical composition of  claim 70 .