MODALITIES FOR THE TREATMENT OF DEGENERATIVE DISEASES OF THE RETINA

• US 20100299765A1
• Filed: 05/18/2010
• Published: 11/25/2010
• Est. Priority Date: 01/23/2004
• Status: Active Grant

• Current Assignee: President and Fellows of Harvard College (Harvard Corporation)
• Original Assignee: Ocata Therapeutics, Inc. (Astellas Pharma Incorporated)
• Inventors: Klimanskaya, Irina, Lanza, Robert
• Granted Patent: US 9,040,770 B2
• Time in Patent Office: Days
• US Class Current: 800/3
• 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.

8-26. -26. (canceled)

27. A highly purified culture of human RPE cells for transplantation derived from human ES cells under conditions in which the culture for transplantation does not contain any human ES cells, wherein the RPE cells are Pax6- and bestrophin+ and exhibit a characteristic cobblestone, polygonal, epithelial-like appearance and comprise brown pigment dispersed within their cytoplasm.

28. A method for producing an enriched population of human retinal pigment epithelium (RPE) cells, human RPE-like cells, or human RPE progenitor cells, the method comprising:

(a) providing a multilayer population of human embryonic stem (hES) cells or an embryoid body comprising hES cells;

(b) culturing said multilayer population of hES cells or said embryoid body comprising hES cells for a sufficient time to allow for the appearance cells that comprise brown pigment dispersed within their cytoplasm;

(c) selecting one or more of said cells that comprise brown pigment dispersed within their cytoplasm from the culture of step (b) to obtain human retinal pigment epithelium (RPE) cells, human RPE-like cells, or human RPE progenitor cells; and

(d) culturing said human retinal pigment epithelium (RPE) cells, human RPE-like cells, or human RPE progenitor cells obtained in step (c), thereby producing an enriched population of human retinal pigment epithelium (RPE) cells, human RPE-like cells, or human RPE progenitor cells.

29. The method of claim 28, wherein said culturing in step (b) comprises culturing in media selected from the group consisting of:

media lacking exogenously added FGF;

media lacking exogenously added FGF and lacking exogenously added LIF; and

media lacking exogenously added FGF and lacking exogenously added LIF and lacking exogenously added PLASMANATE®

.

30. The method of claim 28, wherein the duration of culturing in step (b) is selected from the group consisting of:

about 6 weeks, between about 6 weeks and about 8 weeks, and between about 3 months and about 5 months.

31. The method of claim 28, wherein step (d) comprises culturing said human retinal pigment epithelium (RPE) cells, human RPE-like cells, or human RPE progenitor cells for a sufficient time for the appearance of cells that are Pax6- and bestrophin+ and exhibit a characteristic cobblestone, polygonal, epithelial-like appearance and comprise brown pigment dispersed within their cytoplasm.

32. The method of claim 28, wherein prior to step (b) said hES cells are cultured in media selected from the group consisting of:

media comprising exogenously added FGF;

media comprising exogenously added FGF and LIF; and

media comprising exogenously added FGF and PLASMANATE®

.