Recombinant AAVS having useful transcytosis properties
The disclosure in some aspects relates to recombinant adeno-associated viruses having distinct tissue targeting capabilities. In some aspects, the disclosure relates to gene transfer methods using the recombinant adeno-associate viruses. In some aspects, the disclosure relates to isolated AAV capsid proteins and isolated nucleic acids encoding the same.
1. A recombinant AAV (rAAV) comprising (i) an AAV capsid protein that is encoded by an isolated nucleic acid comprising a sequence selected from the group consisting of: SEQ ID NOs: 10-14; and (ii) at least one transgene.
2. A composition comprising the rAAV of claim 1 .
3. The composition of claim 2 further comprising a pharmaceutically acceptable carrier.
4. A method for delivering a transgene to a subject comprising administering the rAAV of claim 1 to a subject, wherein the rAAV infects cells of a target tissue of the subject.
5. The method of claim 4 , wherein the at least one transgene encodes a protein, a CNS-protein, or a secreted tumor suppressor protein.
6. The method of claim 5 , wherein the CNS-protein is aspartoacylase (ASPA), or wherein the secreted tumor suppressor protein is IGFBP7 or SRPX.
7. The method of claim 4 , wherein the transgene encodes an immunoglobulin heavy chain or light chain or fragment thereof.
8. The method of claim 4 , wherein the at least one transgene encodes a siRNA, antisense nucleotide, miRNA, miRNA sponge, or TuD RNA.
9. The method of claim 8 , wherein the miRNA is expressed in a cell of the target tissue.
10. The method of claim of claim 4 , wherein the target tissue is skeletal muscle, heart, liver, pancreas, brain or lung.
11. The method of claim 4 , wherein the transgene expresses a transcript that comprises at least one binding site for a miRNA, wherein the miRNA inhibits activity of the transgene, in a tissue other than the target tissue, by hybridizing to the binding site.
12. The method of claim 4 , wherein the transgene comprises a tissue specific promoter or inducible promoter.
13. The method of claim 12 , wherein the tissue specific promoter is a liver-specific thyroxin binding globulin (TBG) promoter, an insulin promoter, a glucagon promoter, a somatostatin promoter, a pancreatic polypeptide (PPY) promoter, a synapsin-1 (Syn) promoter, a creatine kinase (MCK) promoter, a mammalian desmin (DES) promoter, a α-myosin heavy chain (a-MHC) promoter, or a cardiac Troponin T (cTnT) promoter.
14. The method of claim 4 , wherein the rAAV is administered intravenously, intravascularly, transdermally, intraocularly, intrathecally, orally, intramuscularly, subcutaneously, intranasally, or by inhalation.
15. The method of claim 4 , wherein the subject is selected from a human, a mouse, a rat, a rabbit, a dog, a cat, a sheep, a pig, and a non-human primate.
16. A method for treating Canavan disease, the method comprising administering to a subject a therapeutically effective amount of a rAAV to a subject, wherein the rAAV comprises:
(i) a capsid protein that is encoded by an isolated nucleic acid comprising having a sequence selected from SEQ ID NOs: 10 to 14, and
(ii) at least one transgene, wherein the at least one transgene encodes ASPA;
and wherein the rAAV infects cells of a target tissue of the subject.
17. A method for treating cancer, the method comprising administering to a subject a therapeutically effective amount of a rAAV to a subject, wherein the rAAV comprises:
(i) a capsid protein that is encoded by an isolated nucleic acid comprising having a sequence selected from SEQ ID NOs: 10 to 14, and
(ii) at least one transgene,
wherein the at least one transgene encodes (a) a tumor suppressor protein or (b) a siRNA, a microRNA, or an antisense nucleotide that inhibits the expression of a gene product associated with cancer;
and wherein the rAAV infects cells of a target tissue of the subject.
18. The composition of claim 1 , wherein the transgene encodes a peptide, a protein, a siRNA, a microRNA, or an antisense nucleotide.
19. The composition of claim 18 , wherein the transgene encodes a siRNA, a microRNA, or an antisense nucleotide that inhibits the expression of a gene product associated with cancer.
20. The composition of claim 19 , wherein the gene product associated with cancer is SOD1.