Transgenic mouse expressing human lipoprotein (a) with disabled vitamin c gene and its use as a disease treatment model

Abstract

The invention discloses novel model of transgenic mammal, a method of crossbreeding transgenic mammal and the use of the transgenic mammal for assessing prevention and/or treatment methods for cardiovascular and other diseases related to lipoprotein(a). The transgenic mammal expresses human apolipoprotein (a) (apo(a)) and human apolipoprotein B-100 (apo B-100) genes and produces human lipoprotein (a), apo (a) and apo B-100 and produces no vitamin C. This novel dual transgenic mammal is the ideal model for testing pharmaceutical compounds for efficacy and usefulness in the prevention and/or treatment of human diseases.

What is claimed is: 
     
         1 . A dual transgenic mammal, comprising:
 a third strain of transgenic mammal was obtained by crossbreeding a first knockout strain mammal and a second strain of transgenic mammal expressing a set of genes that are human in nature, wherein the first knockout strain mammal is lacking a gulonolactone oxidase gene, GULO gene (GULO−/−), and produces no Vitamin C, wherein the second strain of mammal possesses a human apolipoprotein (a) gene (apo (a)+), wherein the third strain of transgenic mammal expresses the human apo (a) gene (apo (a)+) and, simultaneously lacks the GULO gene (GULO−/−); and   a fifth strain of mammal made by crossbreeding the first knockout strain mammal lacking the GULO gene and the fourth strain of mammal possessing a human apolipoprotein B-100 gene (apo B-100+), wherein the fifth strain of mammal lacks the GULO gene (GULO−/−) and expresses the human apo B-100 gene (apo B-100+).   
     
     
         2 . The transgenic mammal of  claim 1 , further comprising:
 a dual transgenic mammal that lacks the GULO gene (GULO−/−) and, simultaneously, produces human lipoprotein (a) (Lp(a)+) was obtained by crossbreeding the third strain of mammal lacking GULO gene (GULO−/−) and, simultaneously expressing the human apo (a) gene (apo (a)+), and the fifth strain of transgenic mammal lacking the GULO gene (GULO−/−) and, simultaneously, possessing the human apo B-100 gene (apo (B-100)+).   
     
     
         3 . The transgenic mammal of  claim 3 , wherein the first knockout mammal, second, third, fourth, fifth transgenic mammal and the dual transgenic mammal is a mouse. 
     
     
         4 . A method, comprising:
 crossbreeding a first knockout strain and a second strain of mouse to obtain and a third strain of transgenic mouse that do not produce vitamin C and expresses a first human gene to produce the human products;   crossbreeding the first knockout strain and a third strain of mouse to produce a fifth transgenic mouse to express a second human gene and produce no vitamin C; and   crossbreeding the fifth strain of transgenic mouse and the third strain of transgenic mouse to produce novel dual transgenic mouse that expresses the first human gene, the second human gene and a third human gene to make a first product, second product and a third product but produce no vitamin C.   
     
     
         5 . The method of  claim 4 , further comprising:
 testing the transgenic mouse using a specific sequence of primer to confirm the presence of the two human genes, human apo (a) and human apo B-100, the first product, second product, third product and lack of GULO gene.   
     
     
         6 . The method of  claim 4 , wherein the first strain of transgenic mouse lacks the GULO gene (GULO−/−), the second strain of transgenic mouse expresses human apo (a) (+), the third strain lacks the GULO gene (GULO−/−) and expresses human apo (a) (apo (a)+), the fourth strain of transgenic mouse expresses human apo B-100 (B-100+), the fifth strain lacks the GULO gene (GULO−/−) and, simultaneously, expresses human apo B-100 (B-100+). 
     
     
         7 . The method of  claim 5 , wherein the specific sequences are Seq 1 to Seq 12. 
     
     
         8 . The method of  claim 4 , further comprising:
 crossbreeding the third strain expressing human GULO (GULO−/−) and human (apo (a)+), and the fifth strain of transgenic mouse expressing human GULO (GULO−/−) and human apo B-100(B-100+) to obtain a dual transgenic mouse.   
     
     
         9 . The method of  claim 9 , wherein the dual transgenic mouse lack the ability to express the gulonolactone oxidase gene [GULO (GULO−/−)] and produces human lipoprotein (a) (Lp(a)). 
     
     
         10 . The method of  claim 6 , wherein the first product is human apo (a), second product is human apo B-100 and the third product is human Lp(a). 
     
     
         11 . The method of  claim 9 , further comprising:
 testing an effect of a pharmaceutical drug that will affect Lp(a) synthesis and/or Lp(a) plasma levels and/or Lp(a) deposition inside the vascular wall in the transgenic mouse with the purpose to treat a cardiovascular disease.   
     
     
         12 . A method, comprising:
 crossbreeding a dual transgenic mouse to produce human Lp(a) and not produce Vitamin C due to lack of GULO (GULO−/−) gene using transgenic mouse having a first knockout strain, a second strain to make a third strain and using the first knockout strain and fourth strain to make a fifth strain, using the third strain and the fifth strain to make a dual transgenic mouse; and   treating the dual transgenic mouse with Lp(a)-modulating compounds in order to identify preventive and/or therapeutic approaches for a human Lp(a)-related diseases.   
     
     
         13 . The method of  claim 12 , wherein the first knockout strain of transgenic mouse lacking the GULO gene (GULO−/−), the second strain of transgenic mouse expresses human apo (a) (apo(a)+), the third strain lacks the GULO gene (GULO−/−) and, simultaneously, possesses human apo(a) (apo (a)+) gene, the fourth strain of transgenic mouse possesses human apo B-100 (B-100+), the fifth strain lacks the GULO gene (GULO−/−) and, simultaneously, expresses human apo B-100 (B-100+). 
     
     
         14 . The method of  claim 13 , wherein the first knockout strain does not produce Vitamin C. 
     
     
         15 . The method of  claim 13 , wherein the second strain of transgenic mouse produces human apo (a). 
     
     
         16 . The method of  claim 13 , wherein the third strain of transgenic mice does not produce Vitamin C and produces human apo (a). 
     
     
         17 . The method of  claim 13 , wherein the fifth strain of transgenic mouse does not produce Vitamin C and produces human apo B-100. 
     
     
         18 . The method of  claim 12 , wherein the dual transgenic mouse lacks the GULO gene and does not produce Vitamin C and, simultaneously produces human Lp(a). 
     
     
         19 . The method of  claim 12 , wherein the human Lp(a)-related disease is a cardiovascular disease. 
     
     
         20 . The method of  claim 12 , wherein the human Lp(a)-related disease is inflammatory, infectious or degenerative in nature.