Chimeric vectors
The present invention is based, in part, on the discovery that parvovirus (including AAV) capsids can be engineered to incorporate small, selective regions from other parvoviruses that confer desirable properties. The inventors have discovered that in some cases as little as a single amino acid insertion or substitution from a first parvovirus (e.g., an AAV) into the capsid structure of another parvovirus (e.g., an AAV) to create a chimeric parvovirus is sufficient to confer one or more of the desirable properties of the first parvovirus to the resulting chimeric parvovirus and/or to confer a property that is not exhibited by the first parvovirus or is present to a lesser extent.
1. A method of administering a nucleic acid to a cell comprising contacting the cell with a chimeric virus vector comprising:
a) a chimeric adeno-associated virus (AAV) capsid comprising a selective amino acid insertion immediately following amino acid position 264 in an AAV2 capsid subunit having the amino acid sequence of (SEQ ID NO:3) or a corresponding change in another AAV capsid subunit; and
b) a nucleic acid comprising an AAV terminal repeat sequence and a heterologous nucleic acid sequence,
wherein the nucleic acid is packaged within the chimeric AAV capsid.
2. A method of delivering a nucleic acid to a subject comprising administering to the subject a chimeric virus vector comprising:
a) a chimeric adeno-associated virus (AAV) capsid comprising a selective amino acid insertion immediately following amino acid position 264 in an AAV2 capsid subunit having the amino acid sequence of (SEQ ID NO:3) or a corresponding change in another AAV capsid subunit; and
b) a nucleic acid comprising an AAV terminal repeat sequence and a heterologous nucleic acid sequence,
wherein the nucleic acid is packaged within the chimeric AAV capsid.
3. The method of claim 2 , wherein the subject is a human subject.
4. The method of claim 2 , wherein the subject has or is at risk for a disorder selected from the group consisting of a muscular dystrophy including Duchenne or Becker muscular dystrophy, hemophilia A, hemophilia B, multiple sclerosis, diabetes mellitus, Gaucher disease, Fabry disease, Pompe disease, cancer, arthritis, muscle wasting, heart disease including congenital heart failure or peripheral artery disease, intimal hyperplasia, a neurological disorder including epilepsy, Huntington's disease, Parkinson's disease or Alzheimer's disease, an autoimmune disease, cystic fibrosis, thalassemia, Hurler's disease, Krabbe's disease, phenylketonuria, Batten's disease, spinal cerebral ataxia, LDL receptor deficiency, hyperammonemia, anemia, arthritis, a retinal degenerative disorder including macular degeneration, adenosine deaminase deficiency, and cancer including tumor-forming cancers.
5. The method of claim 2 , wherein the chimeric virus vector or pharmaceutical formulation is administered to skeletal muscle.
6. The method of claim 5 , wherein the subject has or is at risk for muscular dystrophy.
7. The method of claim 5 , wherein the chimeric virus vector comprises a chimeric AAV capsid comprising a selective amino acid insertion following amino acid position 264 in an AAV2 capsid subunit.
8. The method of claim 5 , wherein the chimeric virus vector comprises a chimeric AAV capsid comprising a selective amino acid insertion following amino acid position 264 in an AAV3b capsid subunit.
9. The method of claim 2 , wherein the chimeric virus vector or pharmaceutical formulation is administered to cardiac muscle.
10. The method of claim 9 , wherein the subject has or is at risk for heart disease.
11. The method of claim 10 , wherein the subject has or is at risk for congestive heart failure or peripheral artery disease.
12. The method of claim 10 , wherein the chimeric virus vector comprises a chimeric AAV capsid comprising a selective amino acid insertion following amino acid position 264 in an AAV3b capsid subunit.
13. The method of claim 2 , wherein the chimeric virus vector or pharmaceutical formulation is administered to glial cells and/or astrocytes.
14. The method of claim 13 , wherein the subject has or is at risk for Parkinson's Disease, astrocytomas or glioblastoma.
15. The method of claim 13 , wherein the chimeric virus vector comprises a chimeric AAV capsid comprising a selective amino acid insertion following amino acid position 264 in an AAV2 capsid subunit.