Models of thrombotic thrombocytopenic purpura and methods of use thereof
View Patent ↗The invention relates to the development of an animal model for testing various agents in the treatment of a clotting disorder. More specifically, the invention relates to the use of ultra-large molecular weight multimers of von Willebrand factor (VWF) in various mouse strains to induce thrombotic thrombocytopenic purpura (TTP)-like symptoms for the development of a mouse model of TTP. The invention also provides methods for generating such animal disease models and screening methods for identifying biologically active compounds which are effective in the treatment of TTP.
1. A mouse model for testing the efficacy of a therapeutic agent in the treatment of a blood clotting disorder characterized by histopathological changes indicative of disseminated intravascular coagulopathy (DIC) or thrombotic thrombocytopenic purpura (TTP), said mouse comprising a circulating level of recombinant human von Willebrand factor (VWF) polypeptide comprising ultra-large VWF multimers in an amount sufficient to induce DIC or microthrombosis caused by administering a non-lethal dosage greater than or equal to 1000 RCoU/kg body weight of recombinant human VWF polypeptide, wherein said mouse has inability to break down recombinant human VWF polypeptide, and wherein said mouse exhibits histopathological changes indicative of DIC and/or TTP.
2. The mouse model of claim 1 , wherein the mouse is deficient in a disintegrin and metalloprotease with thrombospondin type 1 domains 13 (ADAMTS13) polypeptide.
3. The mouse model of claim 1 , wherein the mouse is deficient in VWF polypeptide.
4. The mouse model of claim 1 , wherein the mouse is of a C57BL/6J strain.
5. The mouse model of any one of claims 1 and 2 - 4 , wherein the mouse further comprises recombinant Factor VIII (rFVIII).
6. A transgenic mouse whose genome comprises homozygous disruption of endogenous a disintegrin and metalloprotease with thrombospondin type I domains 13 gene (ADAMTS 13−/−), wherein the mouse further comprises a circulating level of recombinant human von Willebrand factor (VWF) polypeptide comprising ultra-large VWF multimers in an amount sufficient to induce disseminated intravascular coagulopathy (DIC) or microthrombosis caused by administering a non-lethal dosage greater than or equal to 500 RCoU/kg body weight of recombinant human VWF polypeptide, wherein said mouse has an inability to break down recombinant human VWF polypeptide, and wherein said mouse exhibits histopathological changes indicative of DIC and/or thrombotic thrombocytopenic purpura (TTP).
7. The mouse of claim 6 , wherein the mouse is of a C57BL/6J strain.
8. The mouse of claim 6 , wherein the dosage is a non-lethal dosage greater than or equal to 1000 RCoU/kg body weight.
9. The mouse of claim 6 , wherein the dosage is a non-lethal dosage greater than or equal to 2000 RCoU/kg body weight.
10. The mouse of claim 6 , wherein the mouse further comprises recombinant Factor VIII (rFVIII).