# US 11,111,279 B2 · Nato3 mutant polypeptides and uses thereof > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,111,279 B2 - Document ID: US11111279B2 - Status: Granted - Kind: B2 - Application number: 15/776,580 - Filed: 2016-11-18 - Published: 2018-12-06 - Granted: 2021-09-07 - Earliest claimed: 2015-11-20 - Assignees: Grand Valley State University - Applicants: Grand Valley State University - Inventors: Merritt DeLano-Taylor; Jordan Straight; Doug Peterson; Nick Huisingh; Daniel Doyle - Web page: https://patentgrep.com/patent/US11111279B2 ## CPC - C07K 14/4702: {Regulators; Modulating activity} - C12N 5/0619: {Neurons} - A61K 38/00: Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00) - C12N 2501/40: Regulators of development - 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) - 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) ## Abstract The present invention relates to isolated Nato3 mutant polypeptides. Methods for stimulating a brain cell to differentiate into a dopaminergic progenitor neuronal cell or a dopaminergic neuron comprises increasing phosphorylation of Nato3 in the brain cells and culturing the brain cells until a progenitor dopaminergic neuronal cell marker or a dopaminergic neuronal cell marker is expressed in the cultured brain cells. Methods for treating Parkinson's disease (PD) in a subject comprises administering to the subject in need thereof, a composition comprising progenitor dopaminergic neuronal cells and/or dopaminergic neuronal cells expressing a Nato3 mutant polypeptide to the brain of the subject. ## Claims ### Claim 1 An isolated mutant Nato3 polypeptide, the polypeptide comprising at least one mutation in any one or more serine or threonine amino acid residues in the HLH domain defined by amino acids 99 to 158 of SEQ ID NO: 1, wherein the at least one mutation comprises a substitution of serine or threonine with either aspartic acid or glutamic acid. ### Claim 2 (depends on 1) The isolated mutant Nato3 polypeptide according to claim 1, wherein the at least one mutation occurs at position 99, 130, or 138, or combinations thereof of SEQ ID NO: 1. ### Claim 3 (depends on 2) The isolated mutant Nato3 polypeptide according to claim 2, wherein the at least one mutation occurs at position 99. ### Claim 4 (depends on 2) The isolated mutant Nato3 polypeptide according to claim 2, wherein the at least one mutation occurs at position 130. ### Claim 5 (depends on 2) The isolated mutant Nato3 polypeptide according to claim 2, wherein the at least one mutation occurs at position 138. ### Claim 6 An isolated mutant Nato3 polypeptide, the polypeptide comprising at least one mutation in any one or more serine or threonine amino acid residues in the HLH domain defined by amino acids 99 to 158 of SEQ ID NO: 2, wherein the at least one mutation comprises a substitution of serine or threonine with either aspartic acid or glutamic acid. ### Claim 7 (depends on 6) The isolated mutant Nato3 polypeptide according to claim 6, wherein the at least one mutation occurs at position 101, 132, or 140, or combinations thereof of SEQ ID NO: 2. ### Claim 8 (depends on 7) The isolated mutant Nato3 polypeptide according to claim 7, wherein the at least one mutation occurs at position 101. ### Claim 9 (depends on 7) The isolated mutant Nato3 polypeptide according to claim 7, wherein the at least one mutation occurs at position 132. ### Claim 10 (depends on 7) The isolated mutant Nato3 polypeptide according to claim 7, wherein the at least one mutation occurs at position 140. ### Claim 11 (depends on 1, 6) A method of stimulating a population of brain cells to differentiate into dopaminergic progenitor neuronal cells or dopaminergic neuronal cells, the method comprising expressing a Nato3 mutant polypeptide according to claim 1 or 6 in the brain cells and culturing the brain cells until a progenitor dopaminergic neuronal cell marker or a dopaminergic neuronal cell marker is expressed in the cultured brain cells. ### Claim 12 (depends on 11) The method according to claim 11, wherein brain cells are stem cells. ### Claim 13 (depends on 11) The method according to claim 11, wherein brain cells are isolated from the brain floor plate. ### Claim 14 (depends on 1, 6, 11) The method according to claim 11, wherein expressing the Nato3 mutant polypeptide in the brain cells comprises introducing into the brain cells at least one polynucleotide which encodes a mutant Nato3 polypeptide according to claim 1 or 6. ### Claim 15 (depends on 1, 6, 14) The method according to claim 14, wherein expressing the Nato3 mutant polypeptide in the brain cells comprises transfecting or transforming the brain cells with an expression vector which comprises a polynucleotide sequence which encodes a mutant Nato3 polypeptide according to claim 1 or 6. ### Claim 16 (depends on 11) The method according to claim 11, wherein expression of a progenitor dopaminergic neuronal cell marker in the brain cells comprises an expression of at least one marker selected from the group consisting of Shh, Lmx1b, and Foxa2. ### Claim 17 (depends on 11) The method according to claim 11, wherein the dopaminergic progenitor neuronal cells are determined to be differentiated when at least 50% of the population of brain cells express at least one marker selected from the group consisting of Shh, Lmx1b, and Foxa2. ### Claim 18 (depends on 11) The method according to claim 11, wherein the dopaminergic neuronal cells are determined to be differentiated when at least 50% of the population of brain cells express Nurr1. ### Claim 19 (depends on 1, 6) A genetically modified isolated population of dopaminergic progenitor neurons, the isolated population comprising the mutant Nato3 polypeptide according to claim 1 or 6. ### Claim 20 (depends on 1, 6) A method for treating Parkinson's disease (PD) in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a composition comprising the mutant Nato3 polypeptide of claim 1 or 6.