Aav virions with decreased immunoreactivity and uses therefor

Abstract

Methods of making and using recombinant AAV virions with decreased immunoreactivity are described. The recombinant AAV virions include mutated capsid proteins or are derived from non-primate mammalian AAV serotypes and isolates that display decreased immunoreactivity relative to AAV-2.

1 . A mutated adeno-associated virus (AAV) capsid protein that when present in an AAV virion imparts decreased immunoreactivity to the virion as compared to the corresponding wild-type virion. 
     
     
         2 . The protein of  claim 1 , wherein the mutation comprises at least one amino acid substitution, deletion or insertion to the native protein. 
     
     
         3 . The protein of  claim 2 , wherein the mutation comprises at least one amino acid substitution. 
     
     
         4 . The protein of  claim 3 , wherein the at least one amino acid substitution is in the spike or plateau region of the AAV virion surface. 
     
     
         5 . The protein of  claim 4 , wherein the amino acid substitution comprises a substitution of one or more of the amino acids occurring at a position corresponding to a position of the AAV-2 VP2 capsid selected from the group consisting of amino acid 126, 127, 128, 130, 132, 134, 247, 248, 315, 334, 354, 357, 360, 361, 365, 372, 375, 377, 390, 393, 394, 395, 396, 407, 411, 413, 418, 437, 449, 450, 568, 569, and 571. 
     
     
         6 . The protein of  claim 5 , wherein the naturally occurring amino acid at the position is substituted with an alanine. 
     
     
         7 . The protein of  claim 6 , wherein the protein further comprises a substitution of histidine for the amino acid occurring at the position corresponding to the amino acid found at position 360 of AAV-2 VP2. 
     
     
         8 . The protein of  claim 5 , wherein the protein comprises a substitution of lysine for the amino acid occurring at the position corresponding to the amino acid found at position 571 of AAV-2 VP2. 
     
     
         9 . A polynucleotide encoding the mutated protein of  claim 1 . 
     
     
         10 . A polynucleotide encoding the mutated protein of  claim 5 . 
     
     
         11 . A polynucleotide encoding the mutated protein of  claim 6 . 
     
     
         12 . A polynucleotide encoding the mutated protein of  claim 8 . 
     
     
         13 . A recombinant AAV virion comprising the mutated protein of  claim 1 . 
     
     
         14 . A recombinant AAV virion comprising the mutated protein of  claim 5 . 
     
     
         15 . A recombinant AAV virion comprising the mutated protein of  claim 6 . 
     
     
         16 . A recombinant AAV virion comprising the mutated protein of  claim 8 . 
     
     
         17 . The recombinant AAV virion of  claim 13 , wherein said virion comprises a heterologous nucleic acid molecule encoding an antisense RNA or a ribozymes. 
     
     
         18 . The recombinant AAV virion of  claim 13 , wherein said virion comprises a heterologous nucleic acid molecule encoding a therapeutic protein operably linked to control elements capable of directing the in vivo transcription and translation of said protein. 
     
     
         19 . The recombinant AAV virion of  claim 14 , wherein said virion comprises a heterologous nucleic acid molecule encoding a therapeutic protein operably linked to control elements capable of directing the in vivo transcription and translation of said protein. 
     
     
         20 . The recombinant AAV virion of  claim 15 , wherein said virion comprises a heterologous nucleic acid molecule encoding a therapeutic protein operably linked to control elements capable of directing the in vivo transcription and translation of said protein. 
     
     
         21 . The recombinant AAV virion of  claim 16 , wherein said virion comprises a heterologous nucleic acid molecule encoding a therapeutic protein operably linked to control elements capable of directing the in vivo transcription and translation of said protein. 
     
     
         22 . A method of delivering a recombinant AAV virion to a cell or tissue of a vertebrate subject, said method comprising:
 (a) providing a recombinant AAV virion according to  claim 13 ;   (b) delivering said recombinant AAV virion to said cell or tissue, whereby said protein is expressed at a level that provides a therapeutic effect.   
     
     
         23 . The method of  claim 22 , wherein said cell or tissue is a muscle cell or tissue. 
     
     
         24 . The method of  claim 23 , wherein said muscle cell or tissue is derived from skeletal muscle. 
     
     
         25 . The method of  claim 22 , wherein said recombinant AAV virion is delivered into said cell or tissue in vivo. 
     
     
         26 . The method of  claim 25 , wherein said recombinant AAV virion is delivered by intramuscular injection. 
     
     
         27 . The method of  claim 22 , wherein said recombinant AAV virion is delivered into said cell or tissue in vitro. 
     
     
         28 . The method of  claim 22 , wherein said recombinant AAV virion is delivered into the bloodstream. 
     
     
         29 . The method of  claim 28 , wherein said recombinant AAV virion is delivered intravenously. 
     
     
         30 . The method of  claim 28 , wherein said recombinant AAV virion is delivered intraarterially. 
     
     
         31 . The method of  claim 22 , wherein said recombinant AAV virion is delivered to the liver. 
     
     
         32 . The method of  claim 22 , wherein said recombinant AAV virion is delivered to the brain. 
     
     
         33 . A method of delivering a recombinant AAV virion to a cell or tissue of a vertebrate subject, said method comprising:
 (a) providing a recombinant AAV virion, wherein said AAV virion comprises
 (i) a non-primate, mammalian adeno-associated virus (AAV) capsid protein that when present in an AAV virion imparts decreased immunoreactivity to the virion as compared to immunoreactivity of primate AAV-2; and 
 (ii) a heterologous nucleic acid molecule encoding a therapeutic protein operably linked to control elements capable of directing the in vivo transcription and translation of said protein; 
   (b) delivering said recombinant AAV virion to said cell or tissue, whereby said protein is expressed at a level that provides a therapeutic effect.   
     
     
         34 . The method of  claim 33 , wherein said cell or tissue is a muscle cell or tissue. 
     
     
         35 . The method of  claim 34 , wherein said muscle cell or tissue is derived from skeletal muscle. 
     
     
         36 . The method of  claim 33 , wherein said recombinant AAV virion is delivered into said cell or tissue in vivo. 
     
     
         37 . The method of  claim 36 , wherein said recombinant AAV virion is delivered by intramuscular injection. 
     
     
         38 . The method of  claim 33 , wherein said recombinant AAV virion is delivered into said cell or tissue in vitro. 
     
     
         39 . The method of  claim 33 , wherein said recombinant AAV virion is delivered into the bloodstream. 
     
     
         40 . The method of  claim 39 , wherein said recombinant AAV virion is delivered intravenously. 
     
     
         41 . The method of  claim 39 , wherein said recombinant AAV virion is delivered intraarterially. 
     
     
         42 . The method of  claim 33 , wherein said recombinant AAV virion is delivered to the liver. 
     
     
         43 . The method of  claim 33 , wherein said recombinant AAV virion is delivered to the brain.