# US 7,465,582 B1 · Nurr-1 induction of a dopaminergic neuronal fate in a neural stem cell or neural progenitor cell in vitro > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 7,465,582 B1 - Document ID: US7465582B1 - Status: Granted - Kind: B1 - Application number: 09/980,913 - Filed: 2000-04-27 - Granted: 2008-12-16 - Earliest claimed: 2000-04-27 - Assignees: Neuro Therapeutics Limited - Inventors: Ernest Arenas; Thomas Perlmann; Evan Y. Snyder; Joseph Wagner; Peter Akerud - Web page: https://patentgrep.com/patent/US7465582B1 ## CPC - C12N 5/0619: {Neurons} - A61P 25/28: for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia - 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) - C12N 2501/115: Basic fibroblast growth factor (bFGF, FGF-2) - C12N 2501/385: of the family of the retinoic acid recptor, e.g. RAR, RXR; Peroxisome proliferator-activated receptor [PPAR] - C12N 2501/60: Transcription factors - C12N 2502/08: cells of the nervous system - G01N 2500/10: involving cells - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS - 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) - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) ## Abstract The invention relates to the induction of the neuronal fate in neural stem cells or neural progenitor cells. The inventors have found that a neuronal fate in a neural stem cell or neural progenitor cell can be induced by expressing Nurr1 above basal levels within the cell. Nurr1 is a transcription factor of the thyroid hormone/retinoic acid nuclear receptor superfamily. It is shown herein that the expression of Nurr1 above basal levels in neural stem cells or neural progenitor cells increases the proportion of the cells which differentiate toward a neural fate. It has been found that in particular, dopaminergic neural stem cells or progenitor cells by a process including expression of Nurr1 above basal levels in the cells and contact of the cells with one or more factors supplied by or derived from Type I astrocytes of the ventral mesencephalon. ## Claims ### Claim 1 A method of inducing a dopaminergic neuronal fate in a neural stem cell or neural progenitor cell comprising: (a) incubating in vitro a neural stem cell or a neural progenitor cell that overexpresses Nurr-1, wherein Nurr-1 in the cell is overexpressed when compared to an unmodified neural stem cell or neural progenitor cell in vitro under non-pathological conditions, and (b) co-culturing the cell with a Type 1 astrocyte of the ventral mesencepahlon, thereby contacting the cell with one or more factors secreted from said Type 1 astrocyte, whereby the cell that overexpresses Nurr1 is induced to a dopaminergic neuronal fate. ### Claim 2 (depends on 1) The method according to claim 1 comprising additionally contacting the cell with fibroblast growth factor 8 (FGF8). ### Claim 3 (depends on 1) The method according to claim 1 comprising transforming a neural stem cell or a neural progenitor cell with a DNA sequence encoding Nurr1 operatively linked to a promoter. ### Claim 4 (depends on 1) A method according to claim 1 wherein the Type 1 astrocyte is immortalized or is of an astrocyte cell line. ### Claim 5 (depends on 1) A method according to claim 1 wherein said cell is mitotic when it is contacted with said one or more factors. ### Claim 6 (depends on 1) A method according to claim 1 wherein said cell is additionally contacted with one or more agents selected from the group consisting of: basic fibroblast growth factor (bFGF) epidermal growth factor (EGF), an activator of the retinoid X receptor (RXR), and 9-cis retinol. ### Claim 7 (depends on 1) A method according to claim 1 wherein said cell is additionally contacted with a member of the fibroblast growth factor (FGF) family of growth factors. ### Claim 8 (depends on 7) A method according to claim 7 wherein said cell is contacted with bFGF or EGF, and SR11237. ### Claim 9 (depends on 1) A method according to claim 1 wherein the neural stem cell or neural progenitor cell is pretreated with bFGF and/or EGF prior to contacting the cell with one or more factors secreted from a Type 1 astrocyte of the ventral mesencephalon. ### Claim 10 (depends on 1) A method according to claim 1 further comprising formulating a dopaminergic neuron produced by the method into a composition comprising one or more additional components. ### Claim 11 (depends on 10) A method according to claim 10 wherein the composition comprises a pharmaceutically acceptable excipient. ### Claim 12 (depends on 1) The method of claim 1 whereby 5% to 70% of the cells that overexpress Nurr 1 are induced to a dopaminergic neuronal fate.