Animal models of cancer

Abstract

The present invention provides transgenic, large non-human animal models of cancer, as well as methods of using such animal models in the identification and characterization of therapies for cancer.

1 - 185 . (canceled) 
     
     
         186 . A transgenic swine comprising:
 (a) a mutation of an endogenous p53 gene, wherein the p53 mutation is a missense mutation and results in an amino acid substitution corresponding to an R167H amino acid substitution, or a substitution corresponding to amino acid position Y220, G245, R248, or R273; wherein said amino acid substitution results in an altered expression of a p53 translation product and/or in expression of a non-functional p53 protein that corresponds to an alteration in a human p53 gene associated with a lymphoma, a renal tumor, a mesenteric tumor or an osteogenic tumor; and   (b) a mutation of an endogenous KRAS gene, wherein the KRAS mutation is an activating mutation.   
     
     
         187 . The transgenic swine of  claim 186 , comprising the KRAS mutation in both alleles of the endogenous KRAS gene. 
     
     
         188 . The transgenic swine of  claim 186 , wherein the KRAS mutation results in a G12D mutation of the KRAS protein. 
     
     
         189 . The transgenic swine of  claim 186 , wherein the KRAS mutation is from an insertion of a puromycin resistance cassette driven by a phosphoglycerate kinase promoter, wherein the puromycin resistance cassette is flanked by loxP sites. 
     
     
         190 . The transgenic swine of  claim 186 , obtained by a method comprising use of a polynucleotide comprising the nucleic acid sequence of SEQ ID NO:3 or SEQ ID NO:4. 
     
     
         191 . The transgenic swine of  claim 186 , wherein said p53 mutation is a gain of function mutation. 
     
     
         192 . The transgenic swine of  claim 186 , wherein said p53 mutation is present in both alleles of the endogenous p53 gene. 
     
     
         193 . The transgenic swine of  claim 186 , obtained by a method comprising use of a polynucleotide comprising the nucleic acid sequence of SEQ ID NO:1. 
     
     
         194 . The transgenic swine of  claim 186 , comprising a phenotype selected from the group consisting of:
 (a) one or more detectable neoplastic lesion;   (b) defective production of a p53 translation product;   (c) defective production of an altered p53 protein;   (d) defective biological activity of a p53 translation product; and   (e) defective biological activity of an altered p53 protein.   
     
     
         195 . The transgenic swine of  claim 194 , wherein the phenotype is the presence of a lymphoma, an osteogenic tumor, a mesenteric tumor, or a renal tumor. 
     
     
         196 . The transgenic swine of  claim 186  wherein said swine is selectable marker-free. 
     
     
         197 . A method of identifying a therapeutic agent that can be used in the treatment or prevention of cancer comprising:
 administering a candidate therapeutic agent to a transgenic swine wherein said transgenic swine comprises   (a) a mutation of an endogenous p53 gene, wherein the p53 mutation is a missense mutation and results in an amino acid substitution corresponding to an R167H amino acid substitution, or a substitution corresponding to amino acid position Y220, G245, R248, or R273; wherein said amino acid substitution results in an altered expression of a p53 translation product and/or in expression of a non-functional p53 protein that corresponds to an alteration in a human p53 gene associated with a lymphoma, a renal tumor, a mesenteric tumor or an osteogenic tumor; and   (b) a mutation of an endogenous KRAS gene, wherein the KRAS mutation is an activating mutation;   
       wherein said transgenic swine has at least one symptom of cancer; and 
       wherein an improvement in at least one symptom of cancer in said transgenic swine indicates that said therapeutic agent is can be used in the treatment or prevention of cancer. 
     
     
         198 . The method of  claim 197  wherein said transgenic swine comprises the KRAS mutation in both alleles of the endogenous KRAS gene. 
     
     
         199 . The method of  claim 197  wherein said transgenic swine comprises a KRAS mutation that results in a G12D mutation of the KRAS protein. 
     
     
         200 . The method of  claim 197  wherein said transgenic swine comprises a KRAS mutation resulting from an insertion of a puromycin resistance cassette driven by a phosphoglycerate kinase promoter, wherein the puromycin resistance cassette is flanked by loxP sites. 
     
     
         201 . The method of  claim 197  wherein transgenic swine comprises a polynucleotide comprising the nucleic acid sequence of SEQ ID NO:3 or SEQ ID NO:4. 
     
     
         202 . The transgenic swine of  claim 186 , wherein said p53 mutation is a gain of function mutation. 
     
     
         203 . The method of  claim 197 , wherein said p53 mutation is present in both alleles of the endogenous p53 gene. 
     
     
         204 . The method of  claim 197 , obtained by a method comprising use of a polynucleotide comprising the nucleic acid sequence of SEQ ID NO:1. 
     
     
         205 . The method of  claim 197 , comprising a phenotype selected from the group consisting of:
 (a) one or more detectable neoplastic lesion;   (b) defective production of a p53 translation product;   (c) defective production of an altered p53 protein;   (d) defective biological activity of a p53 translation product; and   (e) defective biological activity of an altered p53 protein.   
     
     
         206 . The method of  claim 205 , wherein the phenotype is the presence of a lymphoma, an osteogenic tumor, a mesenteric tumor, or a renal tumor. 
     
     
         207 . The method of  claim 197  wherein said swine is selectable marker-free.