# US 2018/0023052 A1 · Modalities for the treatment of degenerative diseases of the retina

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- Number: US 2018/0023052 A1
- Document ID: US20180023052A1
- Status: Published application
- Kind: A1
- Application number: 15/495,917
- Filed: 2017-04-24
- Published: 2018-01-25
- Earliest claimed: 2004-01-23
- Assignees: Astellas Institute for Regenerative Medicine
- Applicants: Astellas Institute for Regenerative Medicine
- Inventors: Irina V. Klimanskaya; Robert P. Lanza
- Web page: https://patentgrep.com/patent/US20180023052A1

## CPC

- C12N 5/0621: {Eye cells, e.g. cornea, iris pigmented cells (photoreceptors C12N5/062)}
- A61K 35/12: Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells (vaccines or medicinal preparations containing antigens or antibodies A61K39/00)
- A61K 35/30: Nerves; Brain; Eyes; Corneal cells; Cerebrospinal fluid; Neuronal stem cells; Neuronal precursor cells; Glial cells; Oligodendrocytes; Schwann cells; Astroglia; Astrocytes; Choroid plexus; Spinal cord tissue
- A61K 35/36: Skin; Hair; Nails; Sebaceous glands; Cerumen; Epidermis; Epithelial cells; Keratinocytes; Langerhans cells; Ectodermal cells (islets of Langerhans A61K35/39)
- A61K 35/44: Vessels; Vascular smooth muscle cells; Endothelial cells; Endothelial progenitor cells
- A61K 35/545: Embryonic stem cells; Pluripotent stem cells; Induced pluripotent stem cells; Uncharacterised stem cells
- A61P 25/16: Anti-Parkinson drugs
- A61P 27/02: Ophthalmic agents
- A61P 27/06: Antiglaucoma agents or miotics
- C12N 5/062: {Sensory transducers, e.g. photoreceptors; Sensory neurons, e.g. for hearing, taste, smell, pH, touch, temperature, pain}
- C12N 2500/32: Amino acids
- C12N 2500/44: Thiols, e.g. mercaptoethanol
- C12N 2501/01: Modulators of cAMP or cGMP, e.g. non-hydrolysable analogs, phosphodiesterase inhibitors, cholera toxin
- C12N 2501/115: Basic fibroblast growth factor (bFGF, FGF-2)
- C12N 2501/15: Transforming growth factor beta (TGF-β)
- C12N 2501/155: Bone morphogenic proteins [BMP]; Osteogenins; Osteogenic factor; Bone inducing factor
- C12N 2501/235: Leukemia inhibitory factor [LIF]
- C12N 2501/33: Insulin
- C12N 2501/60: Transcription factors
- C12N 2506/02: from embryonic cells
- 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

## Abstract

This invention relates to methods for improved cell-based therapies for retinal degeneration and for differentiating human embryonic stem cells and human embryo-derived into retinal pigment epithelium (RPE) cells and other retinal progenitor cells.

## Claims

### Claim 1

A method of treating or preventing retinal degeneration, comprising use of a cell selected from the group consisting of at least one of: RPE cells, RPE-like cells, RPE or RPE-like progenitors derived from mammalian embryonic stem cells.

### Claim 2 (depends on 1)

The method of claim 1, wherein the condition of retinal degeneration is selected from the group consisting of at least one of: retinitis pigmentosa and macular degeneration.

### Claim 3 (depends on 1)

The method of claim 1, further comprising transplantation of the cell by vitrectomy surgery into the subretinal space of the eye.

### Claim 4 (depends on 3)

The method of claim 3, wherein the cells are transplanted in a suspension, matrix, or substrate.

### Claim 5 (depends on 2)

The method of claim 2, wherein the retinitis pigmentosa is associated with an animal model.

### Claim 6 (depends on 5)

The method of claim 5, where in the animal model is selected from the group consisting of: rd mouse, RPE-65 knockout mouse, tubby-like mouse, RCS rat, Abyssinian cat, cone degeneration “cd” dog, progressive rod-cone degeneration “prcd” dog, early retinal degeneration “erd” dog, rod-cone dysplasia 1, 2 & 3 “rcd1, rcd2 and rcd3” dogs, photoreceptor dysplasia “pd” dog, and Briard “RPE-65” dog.

### Claim 7 (depends on 6)

The method of claim 6, wherein the outcome of the therapy in the animal model is evaluated using one or more of behavioral tests, fluorescent angiography, histology, and functional testing such as measuring the ability of the cells to perform phagocytosis (photoreceptor fragments), vitamin A metabolism, tight junctions conductivity, or evaluation using electron microscopy.

### Claim 8

A method for the spontaneous differentiation of hES cells into cells RPE, RPE-like, or RPE progenitor cells, said method comprising:
a) allowing hES cell cultures to overgrow on MEF;
b) allowing the hES cell cultures to form a thick multilayer of cells;
c) culturing the hES cells;
d) isolation and culture of the pigmented RPE, RPE-like, and/or RPE progenitor cells from the resultant cell cultures.

### Claim 9 (depends on 8)

The method of claim 8, wherein the isolation and culture of the RPE-like cells comprises:
a) digesting the cultured hES cells or EB's with an enzyme;
b) selectively isolating the pigmented cells;
c) plating the isolated cells on gelatin or laminin for 1-2 days to form primary cultures (P0);
d) continued culture of the primary culture for a period of up to 3 weeks; and,
e) isolation of the RPE-like cells.

### Claim 10 (depends on 9)

The method of claim 9, where in the enzyme is selected from the group consisting of one or more of trypsin, collagenase, and dispase.

### Claim 11 (depends on 8)

The method of claim 8, wherein the RPE cells are grown to establish a new RPE cell line.

### Claim 12 (depends on 11)

The method of claim 11, wherein the RPE cell line is differentiated into alternate lineages comprising treatment of the RPE cell line in culture with bFGF or FGF.

### Claim 13 (depends on 11)

The method of claim 11, wherein the new RPE cell lines varies from the already established RPE cell lines in at least one of the characteristics selected from the group consisting of: growth rate, expression of pigment, de-differentiation in culture, and re-differentiation in culture, of RPE-like cells when they are derived from different ES cell lines.

### Claim 14

A method for the derivation of RPE lines or precursors to RPE cells that have an increased ability to prevent neovascularization, said method comprising:
a) aging a somatic cell from an animal such that telomerase is shortened wherein at least 10% of the normal replicative lifespan of the cell has been passed; and,
b) using the somatic cell as a nuclear transfer donor cell to create cells that overexpress angiogenesis inhibitors, wherein the angiogenesis inhibitors can be Pigment Epithelium Derived Factor (PEDF/EPC-1).

### Claim 15 (depends on 14)

The method of claim 14, wherein the somatic cells are genetically modified with exogenous genes that inhibit neovascularization.

### Claim 16 (depends on 8)

The method of claim 8, wherein the RPE-like cells are derived from a bank of ES or embryo-derived cells with homozygosity in the HLA region such that ES-derived cells have reduced complexity of their HLA antigens.

### Claim 17 (depends on 8)

The method of claim 8, wherein the ES cells are derived from a human

### Claim 18

(canceled)