Transgenic mice expressing APP-Swedish mutation develop progressive neurologic disease

• US 5,877,399 A
• Filed: 06/17/1996
• Issued: 03/02/1999
• Est. Priority Date: 01/27/1994
• Status: Expired due to Term

• Current Assignee: Johns Hopkins University, University of Minnesota
• Original Assignee: Johns Hopkins University, Regents of The University of Minnesota (University of Minnesota)
• Inventors: Sisodia, Sangram S., Borchelt, David R., Hsiao, Karen
• Primary Examiner(s): Crouch, Deborah
• Application Number:
  US08/664,872
• Time in Patent Office: 988 Days
• Field of Search: 800/2, 800/DIG. 1, 424/9.2, 435/320.1, 536/23.1, 935/60
• US Class Current: 800/3
• CPC Class Codes: A01K 2207/15 Humanized animals
• CPC Class Codes: A01K 2217/00 Genetically modified animals
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A01K 2227/105 Murine
• CPC Class Codes: A01K 2267/0312 Animal model for Alzheimer's disease
• CPC Class Codes: A01K 2267/0318 Animal model for neurodegenerative disease, e.g. non- Alzheimer's
• CPC Class Codes: A01K 2267/0343 Animal model for prion disease
• CPC Class Codes: A01K 2267/0356 Animal model for processes and diseases of the central nervous system, e.g. stress, learning, schizophrenia, pain, epilepsy
• CPC Class Codes: A01K 67/0275 Genetically modified vertebrates, e.g. transgenic
• CPC Class Codes: A01K 67/0278 Knock-in vertebrates, e.g. humanised vertebrates
• CPC Class Codes: 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
• CPC Class Codes: 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
• CPC Class Codes: 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
• CPC Class Codes: C07K 14/4711 Alzheimer's disease; Amyloid plaque core protein
• CPC Class Codes: C12N 15/8509 for producing genetically modified animals, e.g. transgenic
• CPC Class Codes: C12N 2830/008 cell type or tissue specific enhancer/promoter combination

Abstract

Provided is a transgenic non-human eukaryotic animal whose germ cells and somatic cells contain the amyloid precursor protein sequence introduced into the animal, or an ancestor of the animal, at an embryonic stage. In mice, an age-related CNS disorder characterized by agitation, neophobia, seizures, inactivity, diminished cerebral glucose utilization, cortico-limbic gliosis, and death, develops. An acceleration of this disorder occurs in transgenic mice expressing human and mouse Alzheimer amyloid precursor proteins (APP) produced using a hamster prion protein gene-derived cosmid vector that confers position-independent, copy number-dependent expression. In transgenic mice the disorder develops in direct relationship to brain levels of transgenic APP, but mutant APP confers the phenotype at lower levels of expression than wild-type APP. The disorder occurs in the absence of extracellular amyloid deposition, indicating that some pathogenic activities of APP are dissociated from amyloid formation.

Claims

1. A transgenic mouse whose genome comprises:

an amyloid precursor protein transgene comprising regulatory sequences from a prion gene promoter operatively linked to a coding sequence selected from the group consisting of APP695.WT, APP695.SWE and APP695.TRI, wherein neurological expression of said transgene produces impaired performance of said mouse in memory and learning tests and induces abnormal neuropathology in a cortico-limbic region of said mouse'"'"'s brain, wherein said impaired performance and said abnormal neuropathology are in comparison with control mice.

2. The transgenic mouse according to claim 1, wherein said transgene expression produces levels of amyloid percursor protein at least about two-fold that of endogenous amyloid precursor protein in a control mouse.

3. The transgenic mouse according to claim 1, wherein said abnormal neuropathology is thioflavin S positive amyloid plaques.

4. The transgenic mouse according to claim 1, wherein a nontransgenic ancestor of said mouse is from a strain having greater longevity as compared with other strains of mice.

5. Progeny of said transgenic mice according to claim 1, wherein the genome of the progeny comprises an amyloid precursor protein transgene comprising regulatory sequences from a prion gene promoter operatively linked to a coding sequence selected from the group consisting of APP695.WT, APP695.SWE and APP695.TRI, wherein neurological expression of said transgene produces impaired performance of said progeny in memory and learning tests and induces abnormal neuropathology in a cortico-limbic region of said progeny'"'"'s brain, wherein said impaired performance and said abnormal neuropathology are in comparison with control mice.

6. A method for screening for an agent which ameliorates symptoms of Alzheimer'"'"'s disease, said method comprising:

comparing performance on memory and learning tests of a first transgenic mouse contacted with said agent with that of a second transgenic mouse not contacted with said agent, wherein the genome of said first and said second transgenic mice comprise an amyloid precursor protein transgene comprising regulatory sequences from a prion gene promoter operatively linked to a coding sequence selected from the group consisting of APP695.WT, APP695.SWE and APP695.TRI, wherein neurological expression of said transgene produces impaired performance on said memory learning tests and abnormal neuropathology in a cortico-limbic region of said mouse'"'"'s brain, wherein said impaired performance and said abnormal neuropathology are in comparison with control mice, whereby an agent which ameliorates said symptoms is identified by superior performance of said first transgenic mouse in comparison with said second transgenic mouse on said memory and learning tests.

7. A method for screening for an agent useful for treating Alzheimer'"'"'s disease, said method comprising:

comparing performance on memory and learning tests of a first transgenic mouse contacted with said agent with that of a second transgenic mouse not contacted with said agent, wherein the genome of said first and said second transgenic mice comprise an amyloid precursor protein transgene comprising regulatory sequences from a prion gene promoter operatively linked to a coding sequence selected from the group consisting of APP695.WT, APP695.SWE and APP695.TRI, wherein neurological expression of said transgene produces impaired performance on said memory and learning tests and abnormal neuropathology in a cortico-limbic region of said mouse'"'"'s brain, wherein said impaired performance and said abnormal neuropathology are compared with control mice; and

comparing neuropathology in a cortico-limbic region of the brain of said first and said second transgenic mice when said first transgenic mouse exhibits superior performance on said memory and learning tests compared with said second transgenic mouse, whereby an agent which is useful for treating Alzheimer'"'"'s disease is identified by a decrease in neuropathologic findings in said first transgenic mouse in comparison with said second transgenic mouse.

8. The method according to claim 7, wherein said decreased neuropathologic findings are one or more findings selected from the group consisting of:

a reduction in number of thioflavin S-positive Aβ

deposits;

a reduction in amount of thioflavin S-positive Aβ

deposits;

a reduction of hypertrophic gliosis in cortico-limbic structures of said brain;

a reduction of diminution of 2-deoxyglucose uptake in cortico-limbic structures of said brain; and

a reduction of diminution of 2-deoxyglucose utilization in cortico-limbic structures of said brain.

9. A method for screening for an agent useful for treating Alzheimer'"'"'s disease, said method comprising:

comparing performance on memory and learning tests of a first transgenic mouse contacted with said agent with that of a second transgenic mouse not contacted with said agent, wherein the genome of said first and said second transgenic mice comprise an amyloid precursor protein transgene comprising regulatory sequences from a prion gene promoter operatively linked to a coding sequence selected from the group consisting of APP695.WT, APP695.SWE and APP695.TRI, wherein neurological expression of said transgene produces impaired performance on said memory and learning tests and abnormal neuropathology in a cortico-limbic region of said mousers brain, wherein said impaired performance and abnormal neuropathology are in comparison with control mice; and

comparing age of death of said first and said second transgenic mice when said first transgenic mouse exhibits superior performance on said memory and learning tests in comparison with said second transgenic mouse, whereby an agent which is useful for treating Alzheimer'"'"'s disease is identified by a greater age at death of said first transgenic mouse in comparison with said second transgenic mouse.

10. A method for screening for an agent which ameliorates symptoms of Alzheimer'"'"'s disease, said method comprising:

comparing exploratory behavior or locomotor behavior of a first transgenic mouse contacted with said agent with that of a second transgenic mouse not contacted with said agent, wherein the genome of said first and said second transgenic mice comprise an amyloid precursor protein transgene comprising regulatory sequences from a prion gene promoter operatively linked to a coding sequence selected from the group consisting of APP695.WT, APP695.SWE and APP695.TRI, wherein neurological expression of said transgene produces impaired performance on said exploratory and locomotor behavior and abnormal neuropathology in a cortico-limbic region of said mouse'"'"'s brain, wherein said impaired performance and abnormal neuropathology are compared with control mice, whereby an agent which ameliorates said symptoms is identified by less impaired exploratory or locomotor behavior in said first transgenic mouse in comparison with said second transgenic mouse.

11. The method according to claim 10, wherein said exploratory or locomotor behavior is assessed using a corner index test.

12. The method according to claim 10, wherein a nontransgenic ancestor of said mouse is from a strain selected from the group consisting of FVB, Swiss Webster, and C57B6.

13. A transgenic mouse with progressive neurological disease whose genome comprises:

an amyloid precursor protein transgene comprising regulatory sequences from a prion gene promoter operatively linked to an amyloid precursor protein coding sequence having the Swedish mutation, wherein neurological expression of said transgene produces an age-dependent decline in performance of said mouse in memory and learning tests and produces amyloid plaques that are detectable by Congo red staining in the brain of said mouse, wherein said age-dependent decline in performance and said amyloid plaques are in comparison with control mice.