Recombinant adeno-associated viral vectors for delivering a therapeutic gene to lungs
The present invention provides a recombinant adeno-associated virus (rAAV) particle of 6.2 serotype comprising a recombinant nucleic acid comprising a lung-specific SpB promoter and a therapeutic gene, wherein the lung-specific promoter comprises SEQ ID NO: 1, and wherein said promoter is operably linked to and regulates the expression of the therapeutic gene.
1 . A recombinant adeno-associated virus (rAAV) particle comprising:
a) a recombinant nucleic acid comprising a lung-specific SpB promoter and a therapeutic gene, wherein the lung-specific SpB promoter comprises SEQ ID NO: 1, and wherein the lung-specific SpB promoter is operably linked to the therapeutic gene, and
b) an AAV capsid of 6.2 serotype.
2 . The rAAV particle of claim 1 , wherein the therapeutic gene encodes for human telomerase reverse transcriptase (TERT).
3 . The rAAV particle of claim 1 , wherein the therapeutic gene encodes a protein comprising SEQ ID NO: 2 or 3.
4 . The rAAV particle of claim 1 wherein the AAV capsid of 6.2 serotype comprises a VP1 protein comprising SEQ ID NO: 15.
5 . The rAAV particle of claim 1 , wherein the therapeutic gene comprises SEQ ID NO: 8, 9, 11, 12, or 13.
6 . The rAAV particle of claim 5 , wherein the therapeutic gene comprises SEQ ID NO: 11.
7 . The rAAV particle of claim 1 , wherein the recombinant nucleic acid of a) further comprises a Kozak sequence upstream of the therapeutic gene.
8 . The rAAV particle of claim 7 , wherein the Kozak sequence comprises SEQ ID NO: 4.
9 . The rAAV particle of claim 1 , wherein the recombinant nucleic acid of a) further comprises a polyadenylation (Poly-A) signal placed downstream of the therapeutic gene.
10 . The rAAV particle of claim 9 , wherein the Poly-A signal is a bovine growth hormone polyadenylation signal.
11 . The rAAV particle of claim 1 , wherein the recombinant nucleic acid of a) further comprises a 5′-end and 3′-end inverted terminal repeat (ITR) sequences from AAV serotype 2 (AAV2).
12 . The rAAV particle of claim 11 , wherein the 5′-end and 3′-end AAV2 ITR sequences comprise SEQ ID NO: 6 and 7, respectively.
13 . The rAAV particle of claim 1 , wherein the recombinant nucleic acid of a) comprises SEQ ID NO: 10.
14 . A method of treating or preventing a lung disease comprising administering the rAAV particle of claim 1 to a subject in need thereof.
15 . The method of claim 14 , wherein the recombinant nucleic acid of a) comprises SEQ ID NO: 10.
16 . The method of claim 14 , wherein the lung disease is pulmonary fibrosis.
17 . The method of claim 16 , wherein the pulmonary fibrosis is idiopathic pulmonary fibrosis.