KIR 7.1 gene therapy vectors and methods of using the same
The present invention is directed to gene therapy constructs and pharmaceutical compositions for the expression of Kir7.1. The gene therapy constructs include a vector comprising a promoter operably connected to a polynucleotide encoding a Kir7.1 polypeptide. Methods of treating a subject having a condition associated with insufficient expression or function of a Kir7.1 polypeptide are also provided.
1 . A gene therapy vector comprising a VMD2 promoter operably connected to a polynucleotide encoding a Kir7.1 polypeptide, wherein the VMD2 promoter comprises SEQ ID NO: 4 and is active in the retinal pigment epithelium (RPE) in the eye of a subject, and wherein the Kir7.1 polypeptide comprises SEQ ID NO: 1.
2 . The gene therapy vector of claim 1 , wherein the gene therapy vector is a viral vector.
3 . The gene therapy vector of claim 2 , wherein the viral vector is selected from the group consisting of a retroviral vector, an adeno-associated viral (AAV) vector, and an adenoviral vector.
4 . The gene therapy vector of claim 3 , wherein the viral vector is a lentiviral vector.
5 . The gene therapy vector of claim 3 , wherein the viral vector is an adeno-associated viral vector (AAV).
6 . The gene therapy vector of claim 5 , wherein the AAV vector is an AAV2 vector.
7 . The gene therapy vector of claim 2 , wherein the viral vector is a virus particle and comprises a VSV-G envelope protein.
8 . A lentiviral vector or adeno-associated viral (AAV) vector comprising SEQ ID NO: 6 (VMD2-Kir7.1).
9 . A therapeutic composition comprising the gene therapy vector of claim 1 and a pharmaceutically-acceptable carrier.
10 . A method of treating a subject having a condition associated with insufficient expression or function of a Kir7.1 polypeptide comprising subretinally administering between 10 6 and 10 14 copies of the gene therapy vector of claim 1 to at least one eye of the subject, wherein the condition is Leber congenital amaurosis 16 (LCA16), retinitis pigmentosa, or snowflake vitreoretinal degeneration (SVD).
11 . The method of claim 10 , wherein the condition is associated with at least one loss-of-function mutation in a KCNJ13 gene.
12 . The method of claim 11 , wherein the at least one loss-of-function mutation results in a substitution to SEQ ID NO: 1 selected from the group consisting of W53Ter, Q116R, I120T, T153I, R162Q, R166Ter, L241P, E276A, S105I, and G219Ter.
13 . The method of claim 10 , wherein the subject is human.
14 . A method of expressing a Kir7.1 polypeptide in a retinal pigment epithelium (RPE) cell comprising contacting the RPE cell with the gene therapy vector of claim 1 (i) in vitro or (ii) in vivo by subretinal administration of between 10 6 and 10 14 copies of the gene therapy vector.
15 . The method of claim 10 , wherein a c-wave of the eye of the subject is increased after administration of the gene therapy vector in the eye of the subject as compared to the response in the eye of the subject prior to administration of the gene therapy vector.
16 . The method of claim 10 , wherein the vision of the subject is restored.