Human disease modeling using somatic gene transfer

Abstract

The invention provides a system for modeling neurodegenerative and other diseases through somatic gene transfer. In addition, methods of multiple gene transfer, disease analysis and drug testing are provided for.

What is claimed is:  
     
         1 . A method for producing a non-human animal model of a human disease which comprises transferring at least one aberrant form of at least one gene known to be associated with said disease in humans into appropriate tissue of a living non-human animal under conditions which result in the expression of said at least one aberrant gene, wherein said transferring does not require the modification of the germ-line of said living animal.  
     
     
         2 . The method according to  claim 1  wherein said human disease is a neurodegenerative disease.  
     
     
         3 . The method according to  claim 2  wherein said human disease is selected from the group consisting of Alzheimer's Disease, Parkinson's Disease, and Huntington's Disease.  
     
     
         4 . The method according to  claim 3  wherein said at least one gene is an aberrant form of tau.  
     
     
         5 . The method according to  claim 3  wherein said aberrant form of tau is P301L, associated with “fronto-temporal dementia with Parkinson's linked to chromosome 17 (FTDP-17)”.  
     
     
         6 . The method according to  claim 3  wherein said at least one gene is an aberrant form of alpha-synuclein.  
     
     
         7 . The method according to  claim 6  wherein said aberrant form of alpha-synuclein is mutant α-synuclein (A30P), associated with Parkinson's Disease.  
     
     
         8 . The method according to  claim 3  wherein said at least one gene is a mutant amyloid precursor protein (APP), a mutant presenilin-1 (PS1), or combinations thereof, associated with Alzheimer's Disease.  
     
     
         9 . The method according to  claim 1  which comprises identifying a combination of genes relevant to a particular human pathology and somatically transferring combinations of said genes into tissues appropriate to said particular human pathology in a non-human animal model appropriate to said human pathology.  
     
     
         10 . The method according to  claim 1  comprising: 
 (a) controlling the location to which the genes are transferred, that is spatially controlling gene expression of the transferred genes, in the non-human animal model to which said at least one gene is transferred;  
 (b) controlling the temporal effects of transferred genes at specific times in the development of otherwise normal organisms, or in the development of organisms in which germline modifications have previously been made, by selecting the time at which said transferred genes are introduced into said organism, or by controlling the time of expression of said transferred genes;  
 (c) evaluating the effects of expression of combinations of multiple transgenes, which in a germline transgenic non-human animal would be difficult if not impossible to achieve due to diseases which might prevent the animal model from maturing to the age-appropriate state for modeling onset of a particular, complex human disease;  
 (d) increasing the rate for analyzing multiple genes which contribute to complex, multifactorial human diseases by transferring more than a single gene into an appropriate non-human animal model for said disease;  
 (e) testing pharmaceutical agents for their ability to ameliorate specific diseases induced in said non-human animal model;  
 (f) studying specific human pathologies induced in said non-human animal model by inducing said pathology in said animal model by transferring said at least one gene into said animal model;  
 (g) supplementing an existing germline transgenic model with additional somatically provided gene products to modulate the transgenic model;  
 (h) creating a disease condition in an otherwise healthy animal; and combinations of (a)-(h).  
 
     
     
         11 . A non-human animal produced by the method of  claim 1 .  
     
     
         12 . A pharmaceutical identified through testing of pharmaceutical compounds using the non-human animal produced according to  claim 11 .  
     
     
         13 . A method for inducing neurofibrillary tangles in the brain of a non-human animal which comprises injecting into the brain of said animal an effective amount of a gene expression construct encoding tau, alpha-synuclein, presenilin-1, amyloid precursor protein, IL6, or a combination thereof.  
     
     
         14 . A non-human animal produced according to the method of  claim 13 .  
     
     
         15 . A method for inducing behavioral changes in a non-human animal model which comprises somatic administration of at least one gene directly to the brain of said non-human animal, wherein said at least one gene is associated with a human neurodegenerative disease.  
     
     
         16 . The method according to  claim 1  wherein said at least one aberrant form of said at least one gene is transferred by means of an adeno-associated virus.  
     
     
         17 . A method of identifying a pharmacologically active substance that slows, reduces, reverses or eliminates the adverse effects of a degenerative neurological disease, comprising: 
 a. transferring at least one aberrant form of at least one gene known to be associated with said disease in humans into appropriate tissue of a living non-human animal under conditions which result in the expression of said at least one aberrant gene, wherein said transferring does not require the modification of the germ-line of said living animal;    b. repeating step “a” for a sufficient number of living non-human animals to evaluate a desired number of pharmacologically active substances at a desired number of concentrations, combinations, administration regimes, and evaluation schedules;    c. exposing a specified number of said sufficient number of living non-human animals of step “b” to a specific pharmacologically active substance at a desired number of concentrations and administration regimes, and optionally repeating this exposing for each of a desired number of additional specific pharmacologically active substance(s); and    d. evaluating said specific pharmacologically active substance(s) of step “c”;    wherein said evaluating provides information that identifies at least one pharmacologically active substance that slows, reduces or eliminates the adverse effects of a degenerative neurological disease.    
     
     
         18 . The method of  claim 17 , wherein the step “c” exposing begins prior to said transferring of at least one gene of steps “a” and “b”.  
     
     
         19 . The method of  claim 17 , wherein the step “c” exposing begins prior to and continues after said transferring of at least one gene of steps “a” and “b”.  
     
     
         20 . The method of  claim 17 , wherein the step “c” exposing begins after said transferring of at least one gene of steps “a” and “b”.  
     
     
         21 . The method of  claim 17 , wherein the effects of at least one nutrient regime and/or the effects of at least one environmental stressor also are evaluated.  
     
     
         22 . A method of identifying a nutrient regime that slows, reduces, reverses or eliminates the adverse effects of a degenerative neurological disease, comprising: 
 a. transferring at least one aberrant form of at least one gene known to be associated with said disease in humans into appropriate tissue of a living non-human animal under conditions which result in the expression of said at least one aberrant gene, wherein said transferring does not require the modification of the germ-line of said living animal;    b. repeating step “a” for a sufficient number of living non-human animals to evaluate a desired number of pharmacologically active substances at a desired number of concentrations, combinations, administration regimes, and evaluation schedules;    c. exposing a specified number of said sufficient number of living non-human animals of step “b” to a specific pharmacologically active substance at a desired number of concentrations and administration regimes, and optionally repeating this exposing for each of a desired number of additional specific pharmacologically active substance(s); and    d. evaluating said specific pharmacologically active substance(s) of step “c”;    wherein said evaluating provides information that identifies at least one pharmacologically active substance that slows, reduces or eliminates the adverse effects of a degenerative neurological disease.    
     
     
         23 . A method of identifying an environmental inducer that aggravates, induces, or otherwise contributes to the adverse effects of a degenerative neurological disease, comprising: 
 a. transferring at least one aberrant form of at least one gene known to be associated with said disease in humans into appropriate tissue of a living non-human animal under conditions which result in the expression of said at least one aberrant gene, wherein said transferring does not require the modification of the germ-line of said living animal;    b. repeating step “a” for a sufficient number of living non-human animals to evaluate a desired number of pharmacologically active substances at a desired number of concentrations, combinations, administration regimes, and evaluation schedules;    c. exposing a specified number of said sufficient number of living non-human animals of step “b” to a specific environmental inducer at a desired number of concentrations and administration regimes, and optionally repeating this exposing for each of a desired number of additional specific environmental inducer(s); and    d. evaluating said specific environmental inducer(s) of step “c”;    wherein said evaluating provides information that identifies at least one environmental inducer that aggravates, induces, or otherwise contributes to the adverse effects of a degenerative neurological disease.