Methods and animal model for analyzing age-related macular degeneration
AssigneeUniversity of Kentucky Research Foundation
InventorAmbati Jayakrishna
Life of this patent
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Abstract
Methods for testing candidate drugs for treatment of age-related macular degeneration are provided. Ccl2-deficient, and Ccr2-deficient mice are used to determine the effect of candidate drugs and treatments on development of age-ralated macular degeneration. Also provided is a Ccl2-deficient, Ccr2-deficient dual knockout mouse, which is a useful animal model for age-related macular degeneration.
Claims38
A method for testing a candidate drug for treatment or prevention of age-related macular degeneration (AMD) comprising administering the candidate drug to a Ccl2-deficient knockout mouse, a Ccr2-deficient knockout mouse and/or a Ccl2-deficient/Ccr2-deficient dual knockout mouse and analyzing at least one eye for development or regression of drusen and/or lipofuscin accumulation therein, for affect of the candidate drug on Bruch's membrane, affect on retinal degeneration, in at least one eye, and/or affect on choroidal neovascularization of at least one eye.
A method of screening a test compound for potential utility for treatment of age-related macular degeneration, comprising:
- (a) providing a mouse comprising a disrupted Ccl2 and/or CCR2 gene, wherein the mouse is homozygous for the disrupted gene or genes, and wherein the mouse exhibits drusenand/or lipofuscin deposits, retinal degeneration, and/or choroidal neovascularization in at least one eye at about nine to twenty-four months of age compared to a wild-type mouse that does not have the disrupted gene;
- (b) administering the test compound to the mouse;
- (c) determining the effect of the test compound on drusen, lipofuscin deposition, retinal degeneration, or choroidal neovascularization in at least one eye of the mouse;
- and
- (d) correlating the effect of the test compound on drusen, lipofuscin accumulation, retinal degeneration, and/or choroidal neovascularization with a potential utility to treat age-related macular degeneration.
A Ccl2-deficient/CCR2-deficient dual knockout mouse.
A method of monitoring the expression of a Ccr2 gene in at least one eye of a Ccr2−/− mouse comprising
- (1) introducing a plurality of stem cells obtained from a wild type mouse into the Ccr2−/− mouse to obtain a transplanted mouse, wherein said stem cells express wild type Ccr2;
- (2) observing at least one eye of the transplanted mouse for the effect of the wild type Ccr2 gene expression on drusen or lipofuscin deposition, retinal degeneration, or choroidal neovascularization in at least one eye of the transplanted mouse.
A method of monitoring the effects of expression of a Ccl2 gene in at least one eye of a Ccl2−/− mouse comprising
- (1) introducing a plurality of stem cells obtained from a wild type mouse into the Ccl2−/− mouse to obtain a transplanted mouse, wherein said stem cells express wild type Ccl2;
- (2) observing at least one eye of the transplanted mouse for the effect of the wild type Ccr2 gene expression on drusen or lipofuscin deposition, retinal degeneration, or choroidal neovascularization in at least one eye of the transplanted mouse.
A method of monitoring the effects of expression of a Ccl2 gene, Ccr2 gene or both in at least one eye of a Ccl2 deficient, Ccr2 deficient mouse comprising
- (1) introducing a plurality of stem cells obtained from a wild type mouse into the Ccl2 deficient, Ccr2 deficient mouse to obtain a transplanted mouse, wherein said stem cells express wild type Ccl2 and Ccr2;
- (2) observing at least one eye of the transplanted mouse for the effect of the wild type Ccl2 and/or Ccr2 gene expression on drusen or lipofuscin deposition, retinal degeneration, or choroidal neovascularization in at least one eye of the transplanted mouse.
A method of identifying mutations in the Ccl2 gene, Ccr2 gene, Ccl2 promoter, Ccr2 promoter, Ccl2 enhancer, or combinations thereof comprising
- (1) obtaining an AMD DNA library or genomic DNA from a blood sample of an AMD patient;
- (2) screening the AMD DNA library or genomic DNA for sequences that hybridize under high stringency conditions to a wild type Ccl2 gene , Ccr2 gene;
- the ccl2 promoter, the Ccr2 promoter, or the Ccl2 enhancer and
- (3) sequencing the sequences that hybridize to determine the identity of any mutations contained therein.
An expression vector comprising SEQ ID NO. 9.
An expression vector comprising SEQ ID NO. 10.
A method for screening for mutations that potentially cause or affect the development of AMD in a human comprising
- (1) obtaining an AMD DNA library or genomic DNA from a blood sample of an AMD patient;
- (2) screening the AMD DNA library or genomic DNA for sequences that hybridize under high stringency conditions to a wild type C5 receptor gene or C5a receptor gene;
- (3) sequencing the sequences that hybridize to determine the identity of any mutations contained therein.
A method of treating or preventing AMD in a human comprising administering a nucleic acid encoding a wild type human Ccr2 gene, wild type Ccl2 gene or both to at least one eye of the human, wherein the nucleic acid is operably linked to a promoter, or a small molecule that promotes the function of human Ccr2 or Ccl2, and wherein said nucleic acid is expressed in at least one eye.
Dependencies are inferred from the claim text.