Modalities for the treatment of degenerative diseases of the retina

• US 20070031386A1
• Filed: 07/21/2006
• Published: 02/08/2007
• Est. Priority Date: 01/23/2004
• Status: Active Grant

• Current Assignee: President and Fellows of Harvard College (Harvard Corporation)
• Original Assignee: Advanced Cell Technology Incorporated (Astellas Pharma Incorporated)
• Inventors: Klimanskaya, Irina
• Granted Patent: US 7,795,025 B2
• Time in Patent Office: Days
• US Class Current: 424/93.700
• CPC Class Codes: 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
• CPC Class Codes: 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
• CPC Class Codes: A61K 35/44 Vessels; Vascular smooth muscle cells; Endothelial cells; Endothelial progenitor cells
• CPC Class Codes: A61K 9/0048 Eye, e.g. artificial tears
• CPC Class Codes: A61P 25/16 Anti-Parkinson drugs
• CPC Class Codes: A61P 27/00 Drugs for disorders of the senses
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: A61P 9/14 Vasoprotectives; Antihaemorrhoidals; Drugs for varicose therapy; Capillary stabilisers
• CPC Class Codes: C12N 2501/01 Modulators of cAMP or cGMP, e.g. non-hydrolysable analogs, phosphodiesterase inhibitors, cholera toxin
• CPC Class Codes: C12N 2501/115 Basic fibroblast growth factor (bFGF, FGF-2)
• CPC Class Codes: C12N 2501/15 Transforming growth factor beta (TGF-β)
• CPC Class Codes: C12N 2501/155 Bone morphogenic proteins [BMP]; Osteogenins; Osteogenic factor; Bone inducing factor
• CPC Class Codes: C12N 2501/33 Insulin together with transferrin C12N2500/25; Insulin-like growth factors C12N2501/105
• CPC Class Codes: C12N 2506/02 from embryonic cells
• CPC Class Codes: C12N 5/062 Sensory transducers, e.g. photoreceptors; Sensory neurons, e.g. for hearing, taste, smell, pH, touch, temperature, pain
• CPC Class Codes: C12N 5/0621 Eye cells, e.g. cornea, iris pigmented cells photoreceptors C12N5/062

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

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.

2. 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.

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

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

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

6. 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.

7. 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.

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.

9. 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.

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

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

12. 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.

13. 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.

16. The method of claim 8, wherein the RPE-like cells are derived form 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.

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

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).

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

18. A method for the treatment of Parkinson'"'"'s disease comprising the transplantation of RPE-like cells or progenitor cells.