# US 9,434,992 B2 · Diagnosis marker, diagnosis method and therapeutic agent for amyotrophic lateral sclerosis, and animal model and cell model developing amyotrophic lateral sclerosis

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- Number: US 9,434,992 B2
- Document ID: US9434992B2
- Status: Granted
- Kind: B2
- Application number: 14/195,480
- Filed: 2014-03-03
- Published: 2014-07-03
- Granted: 2016-09-06
- Earliest claimed: 2009-08-25
- Assignees: Hiroshima University
- Applicants: Hiroshima University
- Inventors: Hideshi Kawakami; Hirofumi Maruyama; Hiroyuki Morino
- Web page: https://patentgrep.com/patent/US9434992B2

## CPC

- A01K 67/0278: Knock-in vertebrates, e.g. humanised vertebrates
- A61K 31/00: Medicinal preparations containing organic active ingredients
- A61K 49/0008: {Screening agents using (non-human) animal models or transgenic animal models or chimeric hosts, e.g. Alzheimer disease animal model, transgenic model for heart failure}
- A61P 21/00: Drugs for disorders of the muscular or neuromuscular system
- A61P 21/02: Muscle relaxants, e.g. for tetanus or cramps
- A61P 25/00: Drugs for disorders of the nervous system
- A61P 43/00: Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
- C07K 14/47: from mammals
- C12Q 1/6883: for diseases caused by alterations of genetic material
- A01K 2207/15: Humanized animals
- A01K 2217/15: Animals comprising multiple alterations of the genome, by transgenesis or homologous recombination, e.g. obtained by cross-breeding
- A01K 2227/105: Murine
- A01K 2267/0318: Animal model for neurodegenerative disease, e.g. non- Alzheimer's
- C12Q 2600/156: Polymorphic or mutational markers
- C12Q 2600/172: Haplotypes
- A01K: ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- 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
- 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)
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES

## Abstract

Provided are a diagnosis marker, a diagnosis method, and a therapeutic agent suitable for diagnosing and treating amyotrophic lateral sclerosis (ALS). Also provided are an animal model and a cell model suitable for developing a therapeutic agent and a treatment method for ALS. The diagnosis method for ALS includes: an isolation step in which a nucleic acid is isolated from a specimen taken from a subject; a detection step in which bases expressed in a human chromosome 10 optineurin (OPTN) gene region are detected from the isolated nucleic acid; and a determination step in which it is determined whether or not the detected bases are mutated.

## Claims

### Claim 1

A method for diagnosing amyotrophic lateral sclerosis in a human subject, comprising:
(a) obtaining a sample from a human subject suspected of amyotrophic lateral sclerosis,
(b) isolating nucleic acids from the sample,
(c) amplifying DNA fragments from exon 12 of the OPTN (Optineurin) gene by polymerase chain reaction using primers having the sequence of SEQ ID NO: 5 and 6,
(d) determining whether the base in exon 12 of the OPTN at position 13207995 of chromosome 10 is mutated, and
(e) diagnosing that the subject has amyotrophic lateral sclerosis if the base is mutated.

### Claim 2 (depends on 1)

The method according to claim 1, wherein step (d) determines the mutation of the base from cytosine to thymine.

### Claim 3

A method for diagnosing amyotrophic lateral sclerosis in a human subject, comprising:
(a) obtaining a sample from a subject suspected of amyotrophic lateral sclerosis,
(b) isolating nucleic acids from the sample,
(c) amplifying DNA fragments from exon 14 of the OPTN (Optineurin) gene by polymerase chain reaction using primers having the sequence of SEQ ID NO: 7 and 8,
(d) determining whether the base in exon 14 of the OPTN at position 13214104 of chromosome 10 is mutated, and
(e) diagnosing that the subject has amyotrophic lateral sclerosis if the base is mutated.

### Claim 4 (depends on 3)

The method according to claim 3, wherein step (d) determines the mutation of the base from adenine to guanine.

### Claim 5 (depends on 1)

The method according to claim 1, further comprising a step after step (c) and before step (d) that cutting the amplified DNA fragments with a restriction enzyme MseI.