# US 10,869,465 B2 · Transgenic mouse model of retinal vascular disease, method of making, and method of using > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 10,869,465 B2 - Document ID: US10869465B2 - Status: Granted - Kind: B2 - Application number: 16/058,568 - Filed: 2018-08-08 - Published: 2019-02-14 - Granted: 2020-12-22 - Earliest claimed: 2017-08-10 - Assignees: Osaka University - Applicants: Osaka University - Inventors: Yoko Fukushima; Kohji Nishida; Toru Nakano - Web page: https://patentgrep.com/patent/US10869465B2 ## CPC - A01K 67/0275: Genetically modified vertebrates, e.g. transgenic - 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} - C12N 9/12: transferring phosphorus containing groups, e.g. kinases (2.7) - C12Y 207/11001: Non-specific serine/threonine protein kinase (2.7.11.1), i.e. casein kinase or checkpoint kinase - A01K 2217/052: inducing gain of function - A01K 2227/105: Murine - A01K 2267/035: Animal model for multifactorial diseases - 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) - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - C12Y: ENZYMES ## Abstract Provided are a non-human model animal of a retinal vascular disease that can favorably show symptoms similar to those of human retinal vascular diseases such as human diabetic retinopathy, and a method for producing the non-human model animal. In particular, provided are a non-human model animal that is suitable for establishing a method for treating, preventing, or diagnosing retinal edema, which causes highly impaired vision, and a method for producing the non-human model animal. A method for screening a drug for treating and preventing a retinal vascular disease, the method using a non-human model animal, is provided. Provided are a non-human model animal of a retinal vascular disease in which constitutively active Akt is expressed, a method for producing a non-human model animal of a retinal vascular disease in which constitutively active Akt is expressed, and a method for screening a drug for treating or preventing a retinal vascular disease. ## Claims ### Claim 1 A transgenic mouse model of a retinal vascular disease, wherein the mouse model has a genome comprising a nucleic acid sequence encoding a constitutively active Akt mutant protein: (i) comprising an E40K or E17K substitution mutation; or (ii) in which a Plekstrin homology (PH) domain at the N-terminus has been replaced with a myristoylation signal sequence, wherein the mouse model shows at least one symptom selected from the group consisting of retinal edema, retinal hemorrhage, a retinal microaneurysm, and retinal vascular expansion. ### Claim 2 (depends on 1) The transgenic mouse model of a retinal vascular disease according to claim 1, wherein the mouse model has a genome comprising a nucleic acid sequence encoding a constitutively active Akt mutant protein in which a PH domain at the N-terminus has been replaced with a myristoylation signal sequence. ### Claim 3 (depends on 1) The transgenic mouse model of a retinal vascular disease according to claim 1, wherein expression of the constitutively active Akt mutant protein of (i) or (ii) is under the control of a Cre-LoxP system. ### Claim 4 (depends on 1) The transgenic mouse model of a retinal vascular disease according to claim 1, wherein the retinal vascular disease is diabetic retinopathy. ### Claim 5 (depends on 1) A method for screening a test substance for treating a retinal vascular disease, comprising: a) administering a test substance to the transgenic mouse model of a retinal vascular disease according to claim 1, and b) determining the effect of the test substance on the at least one symptom, wherein a decrease in the at least one symptom as compared to a control indicates the test substance treats retinal vascular disease. ### Claim 6 A method for producing a transgenic mouse model of a retinal vascular disease, the method comprising: a) introducing DNA into a fertilized mouse egg, wherein the DNA encodes: (i) a constitutively active Akt mutant protein comprising an E40K or E17K substitution mutation; or (ii) a constitutively active Akt mutant protein in which a Plekstrin homology (PH) domain at the N-terminus has been replaced with a myristoylation signal sequence; and b) implanting the fertilized mouse egg obtained in step a) into a recipient embryo such that a transgenic mouse model of a retinal vascular disease is obtained, wherein the mouse model has a genome comprising a nucleic acid sequence encoding: (i) a constitutively active Akt mutant protein comprising an E40K or E17K substitution mutation; or (ii) a constitutively active Akt mutant protein in which a PH domain at the N-terminus has been replaced with a myristoylation signal sequence, wherein the mouse model shows at least one symptom selected from the group consisting of retinal edema, retinal hemorrhage, a retinal microaneurysm, and retinal vascular expansion. ### Claim 7 (depends on 6) The method for producing a transgenic mouse model of a retinal vascular disease according to claim 6, wherein the mouse model has a genome comprising a nucleic acid sequence encoding a constitutively active Akt mutant protein in which a PH domain at the N-terminus has been replaced with a myristoylation signal sequence, and expression of the constitutively active Akt mutant protein is controlled using 4-hydroxytamoxifen. ### Claim 8 (depends on 7) The method for producing a transgenic mouse model of a retinal vascular disease according to claim 7, wherein the 4-hydroxytamoxifen is administered at a dose of 5 to 50 μg/g weight/day during a desired period between the first day and the fourteenth day after birth. ### Claim 9 (depends on 6) The method for producing a transgenic mouse model of a retinal vascular disease according to claim 6, wherein expression of the constitutively active Akt mutant protein is under the control of a Cre-loxP system. ### Claim 10 A transgenic mouse whose genome comprises a nucleic acid sequence encoding a constitutively active Akt mutant protein: (i) comprising an E40K or E17K substitution mutation; or (ii) in which a Plekstrin homology (PH) domain at the N-terminus has been replaced with a myristoylation signal sequence, wherein the mouse is capable of developing at least one symptom selected from the group consisting of retinal edema, retinal hemorrhage, a retinal microaneurysm, and retinal vascular expansion after inducing expression of the constitutively active Akt mutant protein. ### Claim 11 (depends on 10) A method for screening a test substance for preventing retinal vascular disease, comprising: a) administering a test substance to the transgenic mouse according to claim 10, b) inducing expression of the constitutively active Akt mutant protein; and c) determining the effect of the test substance on preventing the at least one symptom after inducing expression of the constitutively active Alit mutant protein, wherein a delay in progress or decrease in the at least one symptom as compared to a control indicates the test substance prevents retinal vascular disease.