Thrombotic thrombocytopenic purpura (TTP) is a fatal syndrome. Acquired TTP is mainly caused by autoantibodies that inhibit ADAMTS13 enzyme. Plasma exchange is the only effective therapy available to date.
In Aim1 of this proposal, we will reengineer and characterize a series of novel recombinant ADAMTS13 variants that exhibit increased specific activity, but are resistant to inhibition by autoantibodies from patients with acquired TTP. The completion of this aim will provide novel insights into the structure-function relationship of ADAMTS13 and change how we treat TTP today.
In Aim 2, we propose to determine the antigenic binding epitopes at the amino acid resolution using our novel and groundbreaking deuterium exchange coupled with mass spectrometric analysis approaches. In addition, we will determine the pathogenicity of a panel of inhibitory scFV(s) in murine models of arterial thrombosis and TTP established in the laboratory. The information gained from this study may help our understanding of the molecular mechanisms of acquired TTP and the rational designing of novel recombinant ADAMTS13 variants with desired properties (such as resistance to autoantibody inhibition) for future therapy. Finally, in Aim 3, we will test the hypothesis that ectopic expression of wild-type ADAMTS13 and gain-of-function/antibody-resistant ADAMTS13 variants in platelets would target the therapeutic enzyme directly to sites of injury without being inhibited by circulating anti-ADAMTS13 antibodies. Overall, the information obtained from the completion of the proposed study will provide novel insight into the structure-function relationship of ADAMTS13, help our understandings of the mechanisms of acquired TTP, and provide novel tools for potential therapy of such a fatal TTP syndrome.
This project aims to design and assess the biological functions of novel ADAMTS13 variants with increased specific activity but resistance to autoantibodies. We will also determine the antigenic epitopes of ADAMTS13 using novel hydrogen exchange and mass spectrometry and the pathogenicity of a panel of human monoclonal antibodies from patients with acquired thrombotic thrombocytopenic purpura (TTP). Finally, we will develop an ectopic expression of ADAMTS13 and variants in platelets and determine the role of platelet- delivered ADAMTS13 for modulation of arterial thrombosis and TTP phenotype in mouse models. The information obtained from the study will shed new light on the structure-function relationship of ADAMTS13, help understand the molecular mechanism of TTP, and provide novel tools for therapy for acquired TTP.
|Ostertag, Eric M; Bdeir, Khalil; Kacir, Stephen et al. (2016) ADAMTS13 autoantibodies cloned from patients with acquired thrombotic thrombocytopenic purpura: 2. Pathogenicity in an animal model. Transfusion 56:1775-85|
|Ostertag, Eric M; Kacir, Stephen; Thiboutot, Michelle et al. (2016) ADAMTS13 autoantibodies cloned from patients with acquired thrombotic thrombocytopenic purpura: 1. Structural and functional characterization in vitro. Transfusion 56:1763-74|
|Pillai, Vikram G; Bao, Jialing; Zander, Catherine B et al. (2016) Human neutrophil peptides inhibit cleavage of von Willebrand factor by ADAMTS13: a potential link of inflammation to TTP. Blood 128:110-9|
|Zander, Catherine B; Cao, Wenjing; Zheng, X Long (2015) ADAMTS13 and von Willebrand factor interactions. Curr Opin Hematol 22:452-9|
|Casina, Veronica C; Hu, Wenbing; Mao, Jian-Hua et al. (2015) High-resolution epitope mapping by HX MS reveals the pathogenic mechanism and a possible therapy for autoimmune TTP syndrome. Proc Natl Acad Sci U S A 112:9620-5|
|Lu, R-N; Yang, S; Wu, H M et al. (2015) Unconjugated bilirubin inhibits proteolytic cleavage of von Willebrand factor by ADAMTS13 protease. J Thromb Haemost 13:1064-72|
|Zheng, X Long (2015) ADAMTS13 and von Willebrand factor in thrombotic thrombocytopenic purpura. Annu Rev Med 66:211-25|
|Pickens, Brandy; Mao, Yingying; Li, Dengju et al. (2015) Platelet-delivered ADAMTS13 inhibits arterial thrombosis and prevents thrombotic thrombocytopenic purpura in murine models. Blood 125:3326-34|
|Zheng, X Long (2015) ADAMTS13, lucky to have a hydrophobic pocket. Blood 125:1852-3|
|Hanby, Hayley A; Zheng, X Long (2014) Current status in diagnosis and treatment of hereditary thrombotic thrombocytopenic purpura. Hereditary Genet 3:|
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