Novel adeno-associated virus capsid proteins

Abstract

The invention relates to novel adeno-associated virus (AAV) capsid proteins, AAV particles comprising a novel capsid protein, polynucleotides encoding these capsid proteins and AAV vectors expressing these capsid proteins. The invention also relates to methods of making the herein described AAV vectors expressing the novel capsid proteins of the invention and associated therapeutic uses of thereof.

1 .- 52 . (canceled) 
     
     
         53 . An adeno-associated virus (AAV) viral particle comprising:
 I. (a) a capsid protein, wherein the capsid protein comprises an amino acid sequence that is at least 95% identical to: (i) a VP1 region of SEQ ID NO: 84, (ii) a VP2 region of SEQ ID NO: 84, or (iii) a VP3 region of SEQ ID NO: 84, and wherein the capsid protein comprises variable region (VR) VI of SEQ ID NO: 84; and
 (b) a nucleic acid sequence comprising a transgene, wherein the transgene comprises a heterologous gene operably linked to a regulatory sequence that control the expression of the heterologous gene in a host cell; 
 or 
   II. (a) a capsid protein, wherein the capsid protein comprises an amino acid sequence that is at least 97% identical to: (i) a VP1 region of SEQ ID NO: 84, (ii) a VP2 region of SEQ ID NO: 84, or (iii) a VP3 region of SEQ ID NO: 84, and wherein the capsid protein comprises a GBS region, or variable region (VR) VII, or a GH loop of SEQ ID NO: 84; and
 (b) a nucleic acid sequence comprising a transgene, wherein the transgene comprises a heterologous gene operably linked to a regulatory sequence that controls the expression of the heterologous gene in a host cell; 
 or 
   III. (a) a capsid protein, wherein the capsid protein comprises an amino acid sequence that is at least 98% identical to: (i) a VP1 region of SEQ ID NO: 84, (ii) a VP2 region of SEQ ID NO: 84, or (iii) a VP3 region of SEQ ID NO: 84; and
 (b) a nucleic acid sequence comprising a transgene, wherein the transgene comprises a heterologous gene operably linked to a regulatory sequence that controls the expression of the heterologous gene in a host cell. 
   
     
     
         54 . The AAV viral particle of  claim 53 , wherein the capsid protein comprises a GH loop of SEQ ID NO: 84. 
     
     
         55 . The AAV viral particle of  claim 53 , wherein the capsid protein comprises an amino acid sequence that is at least 99% identical to: (i) the VP1 region of SEQ ID NO: 84, (ii) the VP2 region of SEQ ID NO: 84, or (iii) the VP3 region of SEQ ID NO: 84. 
     
     
         56 . The AAV viral particle of  claim 53 , wherein the capsid protein comprises an amino acid sequence that is identical to: (i) the VP1 region of SEQ ID NO: 84, (ii) the VP2 region of SEQ ID NO: 84, or (iii) the VP3 region of SEQ ID NO: 84. 
     
     
         57 . The AAV viral particle of  claim 53 , wherein the AAV viral particle comprises (i) a capsid protein comprising an amino acid sequence that is identical to the VP1 region of SEQ ID NO: 84, (ii) a capsid protein comprising an amino acid sequence that is identical to the VP2 region of SEQ ID NO: 84, and (iii) a capsid protein comprising an amino acid sequence that is identical to the VP3 region of SEQ ID NO: 84. 
     
     
         58 . The AAV viral particle of  claim 53 , wherein the nucleic acid sequence further comprises an AAV inverted terminal repeat (ITR). 
     
     
         59 . The AAV viral particle of  claim 53 , wherein the nucleic acid sequence further comprises a 5′ AAV ITR and a 3′ AAV ITR. 
     
     
         60 . The AAV viral particle of  claim 53 , wherein the heterologous gene encodes a therapeutic protein. 
     
     
         61 . A composition comprising, the AAV viral particle of  claim 53 , and a physiologically compatible carrier. 
     
     
         62 . A vector comprising:
 I. a nucleic acid sequence encoding an adeno-associated virus (AAV) capsid protein, wherein the capsid protein comprises an amino acid sequence that is at least 95% identical to: (i) a VP1 region of SEQ ID NO: 84; (ii) the VP2 region of SEQ ID NO: 84; or (iii) the VP3 region of SEQ ID NO: 84, wherein the capsid protein comprises variable region (VR) VI of SEQ ID NO: 84, and wherein the nucleic acid sequence is operably linked to a heterologous regulatory sequence that controls the expression of the capsid protein in a host cell;
 or 
   II. a nucleic acid sequence encoding an adeno-associated virus (AAV) capsid protein comprising an amino acid sequence that is at least 97% identical to: (i) a VP1 region of SEQ ID NO: 84, (ii) a VP2 region of SEQ ID NO: 84, or (iii) a VP3 region of SEQ ID NO: 84, and wherein the capsid protein comprises a GBS region, or variable region (VR) VII, or a GH loop of SEQ ID NO: 84, and wherein the nucleic acid sequence is operably linked to a heterologous regulatory sequence that controls the expression of the capsid protein in a host cell;
 or 
   III. a nucleic acid sequence encoding an adeno-associated virus (AAV) capsid protein comprising an amino acid sequence that is at least 98% identical to: (i) a VP1 region of SEQ ID NO: 84, (ii) a VP2 region of SEQ ID NO: 84, or (iii) a VP3 region of SEQ ID NO: 84, wherein the nucleic acid sequence is operably linked to a heterologous regulatory sequence that controls the expression of the capsid protein in a host cell.   
     
     
         63 . The vector of  claim 62 , wherein the capsid protein comprises an amino acid sequence that is at least 99% identical to: (i) the VP1 region of SEQ ID NO: 84, (ii) the VP2 region of SEQ ID NO: 84, or (iii) the VP3 region of SEQ ID NO: 84. 
     
     
         64 . The vector of  claim 62 , wherein the capsid protein comprises an amino acid sequence that is identical to: (i) the VP1 region of SEQ ID NO: 84, (ii) the VP2 region of SEQ ID NO: 84, or (iii) the VP3 region of SEQ ID NO: 84. 
     
     
         65 . The vector of  claim 62 , wherein the capsid protein comprises a GH loop of SEQ ID NO: 84. 
     
     
         66 . The vector of  claim 62 , wherein the heterologous regulatory sequence comprises a constitutive promoter, an inducible promoter, or a tissue-specific promoter. 
     
     
         67 . A cultured host cell containing:
 I. a nucleic acid sequence encoding an AAV capsid protein linked to a heterologous amino acid sequence, wherein the capsid protein comprises an amino acid sequence that is at least 95% identical to: (i) a VP1 region of SEQ ID NO: 84; (ii) the VP2 region of SEQ ID NO: 84; or (iii) the VP3 region of SEQ ID NO: 84, and wherein the capsid protein comprises variable region (VR) VI of SEQ ID NO: 84;
 or 
   II. a nucleic acid sequence encoding an AAV capsid protein linked to a heterologous amino acid sequence, wherein the capsid protein comprises an amino acid sequence that is at least 97% identical to: (i) a VP1 region of SEQ ID NO: 84, (ii) a VP2 region of SEQ ID NO: 84, or (iii) a VP3 region of SEQ ID NO: 84, and wherein the capsid protein comprises a GBS region, or variable region (VR) VII, or a GH loop of SEQ ID NO: 84;
 or 
   III. a nucleic acid sequence encoding an AAV capsid protein linked to a heterologous amino acid sequence, wherein the capsid protein comprises an amino acid sequence that is at least 98% identical to: (i) a VP1 region of SEQ ID NO: 84, (ii) a VP2 region of SEQ ID NO: 84, or (iii) a VP3 region of SEQ ID NO: 84.   
     
     
         68 . The cultured host cell of  claim 67 , wherein the capsid protein comprises an amino acid sequence that is at least 99% identical to: (i) the VP1 region of SEQ ID NO: 84, (ii) the VP2 region of SEQ ID NO: 84, or (iii) the VP3 region of SEQ ID NO: 84. 
     
     
         69 . The cultured host cell of  claim 67 , wherein the capsid protein comprises an amino acid sequence that is identical to: (i) the VP1 region of SEQ ID NO: 84, (ii) the VP2 region of SEQ ID NO: 84, or (iii) the VP3 region of SEQ ID NO: 84. 
     
     
         70 . The cultured host cell of  claim 67 , wherein the cultured host cell further contains a functional AAV rep gene. 
     
     
         71 . The cultured host cell of  claim 67 , wherein the cultured host cell is an insect or mammalian cell. 
     
     
         72 . A host cell comprising, the vector of  claim 62 .