Novel Therapeutics for Acquired Thrombotic Thrombocytopenic Purpura

获得性血栓性血小板减少性紫癜的新疗法

基本信息

批准号：
8504051
负责人：
X. Long Zheng
金额：
$ 40.92万
依托单位：
CHILDREN'S HOSP OF PHILADELPHIA
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-06-01 至 2017-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8504051
关键词：
Adult Amino Acids Antibodies Attenuated Autoantibodies Autoimmune Process B-Lymphocytes Binding Biochemical Biological Biological Assay Biological Process Blood Circulation Blood Platelets Blood Vessels Cleaved cell Clinical Coupled Deuterium Diagnosis Ectopic Expression Endothelial Cells Engineering Enzymes Epitopes Exhibits Future Goals Hematopoietic stem cells Hemolytic Anemia Hemostatic function Human Hydrogen Immunoglobulin G Injury Laboratories Left Lentivirus Vector Libraries Light Maps Mass Spectrum Analysis Metalloproteases Microfluidics Modeling Modification Molecular Mus Organ failure Pathogenesis Pathogenicity Patients Phage Display Phenotype Plasma Plasma Exchange Platelet Factor 4 Platelet Glycoproteins Play Prevention Property Recombinants Resistance Resolution Role Series Shiga Toxin Site Site-Directed Mutagenesis Structure-Activity Relationship Surface Syndrome Testing Therapeutic Thrombocytopenia Thrombosis Thrombotic Thrombocytopenic Purpura Thrombus Transgenic Organisms Transplantation Variant design effective therapy gain of function human monoclonal antibodies inhibitor/antagonist insight mortality mouse model novel novel strategies novel therapeutics promoter public health relevance reconstitution targeted delivery therapeutic development therapeutic enzyme tool von Willebrand Factor

项目摘要

DESCRIPTION (provided by applicant): 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.

描述（由申请人提供）：血栓性血小板减少性紫癜（TTP）是一种致命的综合征。获得性TTP主要是由抑制ADAMTS13酶的自身抗体引起的。血浆置换是迄今为止唯一有效的治疗方法。在本提案的目标 1 中，我们将重新设计并表征一系列新型重组 ADAMTS13 变体，这些变体表现出增加的比活性，但对来自获得性 TTP 患者的自身抗体的抑制具有抵抗力。这一目标的完成将为 ADAMTS13 的结构-功能关系提供新的见解，并改变我们今天对待 TTP 的方式。在目标 2 中，我们建议使用我们新颖且突破性的氘交换结合质谱分析方法来确定氨基酸分辨率的抗原结合表位。此外，我们将在实验室建立的动脉血栓形成和 TTP 小鼠模型中确定一组抑制性 scFV 的致病性。从这项研究中获得的信息可能有助于我们了解获得性 TTP 的分子机制，并合理设计具有所需特性（例如对自身抗体抑制的抵抗）的新型重组 ADAMTS13 变体以用于未来的治疗。最后，在目标 3 中，我们将测试以下假设：血小板中野生型 ADAMTS13 和功能获得/抗体抗性 ADAMTS13 变体的异位表达会将治疗酶直接靶向损伤部位，而不会受到循环抗-抗体的抑制。 ADAMTS13 抗体。总体而言，完成本研究所获得的信息将为 ADAMTS13 的结构-功能关系提供新的见解，帮助我们理解获得性 TTP 的机制，并为这种致命性 TTP 综合征的潜在治疗提供新的工具。