High-transduction-efficiency rAAV vectors, compositions, and methods of use
The present invention provides AAV capsid proteins comprising modification of one or a combination of the surface-exposed lysine, serine, threonine and/or tyrosine residues in the VP3 region. Also provided are rAAV virions comprising the AAV capsid proteins of the present invention, as well as nucleic acid molecules and rAAV vectors encoding the AAV capsid proteins of the present invention. Advantageously, the rAAV vectors and virions of the present invention have improved efficiency in transduction of a variety of cells, tissues and organs of interest, when compared to wild-type rAAV vectors and virions.
1. A method for treating a disease or condition, comprising administering a therapeutically effective amount of an rAAV particle comprising an AAV VP3 protein, comprising:
a non-threonine amino acid residue at a position that corresponds to T251, T329, T330, T454, T455, T491, T503, T550, T581, T592, T597, T659, T660, T671, T701, or T716 of the wild-type AAV2 capsid protein of SEQ ID NO:2, wherein the non-threonine amino acid residue is not alanine, serine, or lysine.
2. The method of claim 1 , wherein the disease or condition is cancer, diabetes, autoimmune disease, kidney disease, cardiovascular disease, pancreatic disease, intestinal disease, liver disease, neurological disease, neuromuscular disorder, neuromotor deficit, neuroskeletal impairment, neurological disability, neurosensory dysfunction, stroke, ischemia, eating disorder, α 1 -antitrypsin (AAT) deficiency, Batten's disease, Alzheimer's disease, Huntington's disease, Parkinson's disease, skeletal disease, trauma, or pulmonary disease in a mammal.
3. The method of claim 1 , wherein the step of administering comprises transducing a population of mammalian cells with a composition that comprises an effective amount of the rAAV particle comprising the AAV VP3 protein.
4. The method of claim 3 , wherein the mammalian cells are endothelial, epithelial, vascular, liver, lung, heart, pancreas, intestinal, kidney, muscle, bone, dendritic, cardiac, neural, blood, brain, fibroblast, or cancer cells.
5. The method of claim 1 , wherein the AAV VP3 protein comprises a valine residue at a position that corresponds to T491.
6. The method of claim 5 , wherein the disease or condition is cancer, diabetes, autoimmune disease, kidney disease, cardiovascular disease, pancreatic disease, intestinal disease, liver disease, neurological disease, neuromuscular disorder, neuromotor deficit, neuroskeletal impairment, neurological disability, neurosensory dysfunction, stroke, ischemia, eating disorder, α 1 -antitrypsin (AAT) deficiency, Batten's disease, Alzheimer's disease, Huntington's disease, Parkinson's disease, skeletal disease, trauma, or pulmonary disease in a mammal.
7. The method of claim 6 , wherein the step of administering comprises transducing a population of mammalian cells with a composition that comprises an effective amount of the rAAV particle comprising the AAV VP3 protein wherein the mammalian cells are endothelial, epithelial, vascular, liver, lung, heart, pancreas, intestinal, kidney, muscle, bone, dendritic, cardiac, neural, blood, brain, fibroblast, or cancer cells.
8. The method of claim 1 , wherein the non-threonine amino acid residue of the AAV VP3 protein is valine.
9. The method of claim 1 , wherein the rAAV particle is an AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, or AAV12 serotype particle.
10. The method of claim 1 , wherein the rAAV particle comprises a nucleic acid segment encoding a therapeutic or diagnostic agent.
11. A method comprising administering to a subject an rAAV particle comprising an AAV VP3 protein, wherein the AAV VP3 protein comprises:
a non-threonine amino acid residue at a position that corresponds to T251, T329, T330, T454, T455, T491, T503, T550, T581, T592, T597, T659, T660, T671, T701, or T716 of the wild-type AAV2 capsid protein of SEQ ID NO:2, wherein the non-threonine amino acid residue is not alanine, serine, or lysine.
12. The method of claim 11 , wherein the subject has a disease or condition, wherein the disease or condition is cancer, diabetes, autoimmune disease, kidney disease, cardiovascular disease, pancreatic disease, intestinal disease, liver disease, neurological disease, neuromuscular disorder, neuromotor deficit, neuroskeletal impairment, neurological disability, neurosensory dysfunction, stroke, ischemia, eating disorder, α 1 -antitrypsin (AAT) deficiency, Batten's disease, Alzheimer's disease, Huntington's disease, Parkinson's disease, skeletal disease, trauma, or pulmonary disease.
13. The method of claim 11 , wherein the non-threonine amino acid residue of the AAV VP3 protein is valine.
14. The method of claim 11 , wherein the AAV VP3 protein comprises a valine residue at a position that corresponds to T491 of the wild-type AAV2 capsid protein of SEQ ID NO:2.
15. The method of claim 11 , wherein the rAAV particle is an AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, or AAV12 serotype particle.
16. A method of transducing a population of cells, the method comprising introducing into a population of cells a composition comprising an rAAV particle comprising an AAV VP3 protein, wherein the AAV VP3 protein comprises:
a non-threonine amino acid residue at a position that corresponds to T251, T329, T330, T454, T455, T491, T503, T550, T581, T592, T597, T659, T660, T671, T701, or T716 of the wild-type AAV2 capsid protein of SEQ ID NO:2, wherein the non-threonine amino acid is not alanine, valine, or lysine.
17. The method of claim 16 , wherein the population of cells comprises endothelial, epithelial, vascular, liver, lung, heart, pancreas, intestinal, kidney, muscle, bone, dendritic, cardiac, neural, blood, brain, fibroblast, or cancer cells.
18. The method of claim 16 , wherein the non-threonine amino acid residue of the AAV VP3 protein is valine.
19. The method of claim 16 , wherein the AAV VP3 protein comprises a valine residue at a position that corresponds to T491.
20. The method of claim 16 , wherein the rAAV particle is an AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, or AAV12 serotype particle.
21. The method of claim 1 , wherein the AAV VP3 protein comprises a non-threonine amino acid residue at a position that corresponds to T491 of the wild-type AAV3 capsid protein of SEQ ID NO: 2, and further comprises at least a non-threonine amino acid residue at a position that corresponds to T251, T329, T330, T454, T455, T503, T550, T581, T592, T597, T659, T660, T671, T701, T713, or T716 of the wild-type AAV2 capsid protein of SEQ ID NO:2.