A Global Test of Hemostatic Risk

DESCRIPTION (provided by applicant): Phase I funding for this project supported successful studies that were aimed at demonstrating the feasibility of developing an easy to use, global test in the arena of clinical hemostasis that would be sensitive to antithrombotic drugs, bleeding disorders and thrombotic pathologies. This test comprises a measurement of the rate of thrombin generation and/or total thrombin potential in a non-perturbed, tissue factor-dependent system. Prior to the Phase I study, this test existed as a complicated ten-plus step research based assay that could only be performed by highly skilled and trained technicians. During the Phase I project period we simplified the assay by developing a blood collection tube containing the components necessary for the initiation of coagulation (relipidated tissue factor) and suppression of the contact phase of coagulation (corn trypsin inhibitor). The development of this tube reduces the complexity of the initial steps in the assay since issues of timing along with the formulation and rapid addition of reagents becomes irrelevant. Once the tube was prepared we were able to optimize assay conditions to yield reproducible results that compared nicely to those obtained with the original research-based assay. Additional accomplishments include the development of a stable quench solution, the design and manufacture of a prototype temperature-controlled assay chamber and the initial development of our own thrombin-antithrombin-III (TAT) ELISA. Collectively, the accomplishments associated with the Phase I project period demonstrate our ability to produce all elements necessary for the thrombin-generation assay (TGA). Phase II funding is being requested to support the efforts that are associated with the further refinement, assembly and validation of the complete assay system. This project is devoted to the development of a "Global Test of Hemostatic Risk" that could ultimately identify individuals in the apparently "normal" population who are at risk for the future development of hemostatic complications (thrombosis or bleeding disorders). Furthermore, this test may facilitate the administration and monitoring of individualized therapeutic treatment regimens in the thrombotic and hemorrhagic patient populations.