# US 11,957,115 B2 · Genetically modified mouse expressing human APOE4 and mouse Trem2 p.R47H and methods of use thereof

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- Number: US 11,957,115 B2
- Document ID: US11957115B2
- Status: Granted
- Kind: B2
- Application number: 16/496,261
- Filed: 2018-03-21
- Published: 2020-01-23
- Granted: 2024-04-16
- Earliest claimed: 2017-03-21
- Assignees: The Jackson Laboratory; Indiana University Research and Technology Corporation
- Applicants: The Jackson Laboratory; Indiana University Research and Technology Corporation
- Inventors: Gareth Howell; Michael Sasner; Gregory Carter; Bruce Lamb
- Web page: https://patentgrep.com/patent/US11957115B2

## CPC

- A01K 67/0278: Knock-in vertebrates, e.g. humanised vertebrates
- A01K 67/0275: Genetically modified vertebrates, e.g. transgenic
- C07K 14/70503: {Immunoglobulin superfamily}
- C07K 14/775: Apolipopeptides
- G01N 33/6896: {Neurological disorders, e.g. Alzheimer's disease}
- A01K 2217/056: due to mutation of coding region of the transgene (dominant negative)
- A01K 2217/072: maintaining or altering function, i.e. knock in
- 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 2267/0312: Animal model for Alzheimer's disease
- G01N 2500/00: Screening for compounds of potential therapeutic value
- A01K: ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- 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)
- 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

Genetically modified mice characterized by one or more symptoms or signs associated with expression of human APOE4p and mouse Trem2p and relevant to non-familial late-onset Alzheimer's disease are provided wherein the genome of the mouse includes: 1) a DNA sequence encoding a human APOE4 protein (APOE4p) operably linked to a promoter; and 2) a DNA sequence encoding a mouse Trem2 protein having a mutation p,R47H (Trem2p) operably linked to a promoter, such that the mouse expresses human APOE4p and mouse Trem2p. Methods ace provided for screening for a compound for use in the treatment of Alzheimer's disease using such genetically modified mice.

## Claims

### Claim 1

A mouse whose genome comprises: (a) a genetically modified endogenous apolipoprotein E (apoE) allele comprising a nucleic acid encoding a humanized apolipoprotein E4 APOE4 protein (APOE4p), wherein the nucleic acid encoding the humanized APOE4p comprises exon 1 of the mouse ApoE4 gene, and exons 2-4 of the human APOE4 gene; and (b) a genetically modified endogenous allele comprising a nucleic acid encoding a mouse triggering receptor expressed on myeloid cells 2 (TREM2) protein modified to include a R47H substitution (TREM2p) relative to mouse wild type TREM2, wherein the mouse is homozygous for the modified apoE allele encoding the human APOE4p and is homozygous for the modified apoE allele encoding the mouse TREM2p, and the mouse exhibits one or more signs of non-familial late-onset Alzheimer's disease associated with expression of the human APOE4p and the mouse TREM2p.

### Claim 2 (depends on 1)

The mouse of claim 1, wherein the one or more signs of non-familial late-onset Alzheimer's disease are selected from APOE4-dependent alterations in cholesterol metabolism, cerebrovascular leakage, and inflammation, relative to a control mouse that does not comprise the nucleic acid encoding a human APOE4p and the nucleic acid encoding a mouse TREM2p.

### Claim 3 (depends on 1)

The mouse of claim 1, wherein one or more of the following genes is differentially expressed in the mouse, relative to a control mouse that does not comprise the nucleic acid encoding a human APOE4p and the nucleic acid encoding a mouse TREM2p: Pcsk2, Mapk10, Mapk9, Prkcq, Slc18a2, Plcb2, Slc6a4, Gng2, Prkcq, Akt3, Gnao1, Plcb3, Arrb2, 116, Myh10, Gng2, Cnr1, Stx1a, Arrb2, Unc13a, Cdk5, Calb1, Slc6a4, Gria4, Cnr1, Thy1, Mapk10, Hcn1, Cdk5, Chl1, Il6, Mapk9, Myh10, Uchl1, Cnr1, Amph, Cdk5, Calb1, Chl1, Slc6a4, Chmp2b, Akt3, Mapk10, Mapk9, Casp7, Prkcq, Cnr1, Slc18a2, and Gnao1.

### Claim 4 (depends on 2)

The mouse of claim 2, wherein the one or more signs of non-familial late-onset Alzheimer's disease is APOE4-dependent alterations in cholesterol metabolism.

### Claim 5 (depends on 2)

The mouse of claim 2, wherein the one or more signs of non-familial late-onset Alzheimer's disease is cerebrovascular leakage.

### Claim 6 (depends on 2)

The mouse of claim 2, wherein the one or more signs of non-familial late-onset Alzheimer's disease is inflammation.