# US 2008/0305083 A1 · Gfap-based gene therapy for treatment of retinal diseases

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- Number: US 2008/0305083 A1
- Document ID: US20080305083A1
- Status: Published application
- Kind: A1
- Application number: 12/104,649
- Filed: 2008-04-17
- Published: 2008-12-11
- Earliest claimed: 2004-04-22
- Assignees: Florida Atlantic University
- Inventors: C. Kathleen DOREY; Janet C. BLANKS; Howard PRENTICE
- Web page: https://patentgrep.com/patent/US20080305083A1

## CPC

- C12N 15/86: Viral vectors
- A61K 48/005: {characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered}
- A61K 48/0058: {Nucleic acids adapted for tissue specific expression, e.g. having tissue specific promoters as part of a contruct}
- A61P 27/02: Ophthalmic agents
- C07H 21/04: with deoxyribosyl as saccharide radical
- C07K 14/78: Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]
- A61K 48/00: Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
- A61N 1/0412: {Specially adapted for transcutaneous electroporation, e.g. including drug reservoirs}
- A61N 1/30: Apparatus for iontophoresis, {i.e. transfer of media in ionic state by an electromotoric force into the body}, or cataphoresis
- A61N 1/327: {for enhancing the absorption properties of tissue, e.g. by electroporation}
- A61N 5/062: {Photodynamic therapy, i.e. excitation of an agent}
- C12N 2750/14143: viral genome or elements thereof as genetic vector
- C12N 2830/008: cell type or tissue specific enhancer/promoter combination
- C12N 2830/48: regulating transport or export of RNA, e.g. RRE, PRE, WPRE, CTE
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- C07H: SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS (derivatives of aldonic or saccharic acids C07C, C07D; aldonic acids, saccharic acids C07C59/105, C07C59/285; cyanohydrins C07C255/16; glycals C07D; compounds of unknown constitution C07G; polysaccharides, derivatives thereof C08B; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; sugar industry C13)
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- A61N: ELECTROTHERAPY; MAGNETOTHERAPY; RADIATION THERAPY; ULTRASOUND THERAPY (measurement of bioelectric currents A61B; surgical instruments, devices or methods for transferring non-mechanical forms of energy to or from the body A61B18/00; anaesthetic apparatus in general A61M; incandescent lamps H01K; infrared radiators for heating H05B)

## Abstract

Compositions and methods for reducing neovascularization. Purified nucleic acid constructs and vectors encoding an anti-angiogenic protein operably linked to a GFAP promoter. Vectors can include at least one hypoxia regulated element, enhancer element and silencer element. Gene therapy methods for reducing, delaying or preventing neovascularization based on the nucleic acid constructs and vectors.

## Claims

### Claim at position 1

1-26. (canceled)

### Claim 27

A method of preventing, reducing or delaying neovascularization, the method comprising the steps of:
(a) providing a subject having or at risk of developing neovascularization in a tissue; and
(b) transducing at least one GFAP-expressing cell type of said subject with an expression vector comprising a purified nucleic acid construct comprising:
at least one nucleic acid encoding an anti-angiogenic protein operably linked to a GFAP promoter, wherein expression of said anti-angiogenic protein by said transduced cell prevents, reduces or delays neovascularization in said tissue in said subject.

### Claim 28 (depends on 27)

The method of claim 27, wherein said tissue is an ocular tissue selected from at least one of the group consisting of retina, vitreous and choroid.

### Claim 29 (depends on 27)

The method of claim 27, wherein said anti-angiogenic protein is selected from the group consisting of angiostatin, endostatin and Tubedown-1.

### Claim 30 (depends on 27)

The method of claim 27, wherein said expression vector further comprises a hypoxia-regulated element, and the expression of said anti-angiogenic protein by said cell is increased under hypoxic conditions.

### Claim 31 (depends on 27)

The method of claim 27, further comprising the step of:
(c) irradiating said tissue with a laser beam, in a dosage sufficient to upregulate GFAP promoter expression in said GFAP-expressing cell type, wherein the level of expression of said anti-angiogenic protein driven by said GFAP promoter is increased in an amount sufficient to prevent, reduce or delay neovascularization in said tissue.

### Claim 32

A method of preventing, reducing or delaying neovascularization in an eye, the method comprising the steps of:
(a) providing a subject having or at risk of developing a condition involving neovascularization in a compartment of at least one eye; and
(b) transducing at least one GFAP-expressing cell type in said eye of said subject with an expression vector comprising at least one nucleic acid encoding an anti-angiogenic protein operably linked to a GFAP promoter, wherein expression of said anti-angiogenic protein by said transduced cell prevents, reduces or delays neovascularization in said eye of said subject.

### Claim 33 (depends on 32)

The method of claim 32, wherein said GFAP-expressing cell type is a Muller cell or a glial cell of the retina.

### Claim 34 (depends on 32)

The method of claim 32, further comprising the step of:
(c) irradiating tissue of said eye with a laser beam, light, radiation, Cystatin C, or transpupillary thermography in an amount a dosage sufficient to upregulate GFAP promoter driven expression of a said nucleic acid under control of said GFAP promoters control.

### Claim 35 (depends on 27)

The method of claim 27, wherein the vector is a plasmid.

### Claim 36 (depends on 27)

The method of claim 27, wherein the vector is a viral vector selected from the group consisting of a rAAV vector, an adenoviral vector and a lentiviral vector.

### Claim 37 (depends on 29)

The method of claim 29, wherein said anti-angiogenic protein is angiostatin.

### Claim 38 (depends on 29)

The method of claim 29, wherein said anti-angiogenic protein is endostatin.

### Claim 39 (depends on 31)

The method of claim 31, wherein the source of said irradiating is selected from the group consisting of a laser beam, light, radiation, or transpupillary thermography.

### Claim 40 (depends on 33)

The method of claim 33 wherein said expression vector further comprises a hypoxia-regulated element, and the expression of said anti-angiogenic protein by said Muller cell or glial cell is increased under hypoxic conditions.

### Claim 41 (depends on 40)

The method of claim 40, wherein said anti-angiogenic protein is endostatin.

### Claim 42 (depends on 40)

The method of claim 40, wherein said anti-angiogenic protein is angiostatin.

### Claim 43 (depends on 33)

The method of claim 33, wherein the cell type is a Muller cell and the anti-angiogenic factor under control of the GFAP promoter is selectively expressed in Muller cells in a local tissue environment under adverse conditions conducive to neovascularization, and not in Muller cells in tissue environments under normal conditions.

### Claim 44 (depends on 36)

The method of claim 36 wherein the viral vector is an rAAV vector.

### Claim 45 (depends on 40)

The method of claim 40, wherein the viral vector is an rAAV vector.