Direct Detection and Characterization of Thrombosis In Vivo

体内血栓形成的直接检测和表征

• 批准号: 10201739
• 负责人: BRUCE ALAN SULLENGER
• 金额: $ 73.45万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-22 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10201739
• 关键词: Acute; Address; Affinity; Animal Model; Antidotes; Azides; Binding; Biodistribution; Biological Models; Blood Platelets; Blood Vessels; Blood coagulation; Blood flow; Cardiovascular system; Catheters; Chemistry; Chronic; Coagulation Process; Complement; Complex; Deep Vein Thrombosis; Detection; Development; Disease; Event; Factor IXa; Fibrinolytic Agents; Fluorine; Future; Human; Image; Imaging Device; Individual; Investigation; Ischemic Stroke; Macaca mulatta; Modeling; Molecular Profiling; Mus; Myocardial Infarction; Nature; Nucleic Acid Biochemistry; Open Bite; Patients; Peripheral; Positron-Emission Tomography; Primates; Proteins; Prothrombin; Protocols documentation; Publishing; Pulmonary Embolism; RNA; Radiochemistry; Radiolabeled; Scanning; Specificity; Stenosis; Surgical complication; Technology; Therapeutic; Therapeutic Intervention; Thrombin; Thrombosis; Thrombus; Time; Translating; Venous; Venous Thrombosis; age group; aged; antithrombin III-protease complex; aptamer; base; combinatorial chemistry; design; experience; imaging approach; imaging modality; imaging probe; in vivo; in vivo imaging; innovation; insight; molecular imaging; molecular modeling; mouse model; multidisciplinary; neurovascular; nonhuman primate; novel diagnostics; novel strategies; personalized care; personalized medicine; rapid detection; risk stratification; thrombotic; tool; translation to humans; von Willebrand Factor

PROJECT SUMMARY / ABSTRACT Vascular thrombosis is a major underlying factor in many cardiovascular, neurovascular, and related disorders, and is a significant post-surgical complication. Current imaging modalities used to assess thrombotic events present challenges because they primarily visualize the lack of blood flow rather than detailed information about a thrombus directly, and they cannot distinguish between newly formed and aged blood clots. Our team has previously demonstrated that high affinity and high specificity RNA aptamers can be generated against a number of coagulation proteins including thrombin and factor IXa. In addition, our team has demonstrated that aptamer-antidote pairs can be used as rapid binding-rapid reversal probes for real time detection of thrombi. The studies proposed in this application integrate aptamer-antidote pairs with sensitive total-body positron emission tomography (PET) imaging to develop a new approach in the way patients with thrombotic events can be stratified and subsequently treated. This proposal represents the convergence of multidisciplinary domains of expertise to explore a new team direction that will have a major impact on the field through the following Specific Aims: (1) Develop and evaluate aptamer-antidote pairs to perform rapid imaging of thrombi in mouse models for molecular thrombus profiling in vivo, and (2) Assess efficiency of radiolabeled aptamers for total-body PET in nonhuman primates. Preliminary results have demonstrated that aptamers can rapidly bind thrombin on active thrombi in vivo and that the antidotes can reverse such binding in under 5 minutes. In Specific Aim 1, we will further develop this technology by conducting studies in mice with aptamer-antidote pairs generated to thrombin, von Willebrand Factor, and platelet protein GPIIb/IIIa individually and combined. Findings will be adapted in Specific Aim 2 to the translational rhesus monkey model system using total-body PET that has demonstrated outstanding sensitivity. These investigations propose a new strategy to address characterization of thrombi in vivo and include a multidisciplinary translational team with expertise in nucleic acid biochemistry, combinatorial chemistry, antithrombotic agents, novel diagnostic imaging tools and methods, and a nonhuman primate model system of profound translational importance.

项目总结/摘要 血管血栓形成是许多心血管、神经血管和相关疾病的主要潜在因素， 并且是严重的术后并发症。目前用于评估血栓形成事件的成像方式 因为它们主要是可视化血流的缺乏，而不是详细的信息 他们无法直接分辨新形成的血栓和老化的血栓。我们的团队 先前已经证明，可以产生高亲和力和高特异性的RNA适体， 包括凝血酶和凝血因子IXa在内的多种凝血蛋白。此外，我们的团队已经证明， 适体-解毒剂对可用作血栓真实的实时检测的快速结合-快速逆转探针。 本申请中提出的研究将适体-解毒剂对与敏感的全身正电子结合起来 发射断层扫描（PET）成像，以开发一种新的方法在血栓事件患者 可以分层并随后进行处理。这一建议代表了多学科的融合 专业领域，探索新的团队方向，通过 以下具体目的：（1）开发和评估适体-解毒剂对，以进行血栓的快速成像， 用于体内分子血栓谱分析的小鼠模型，和（2）评估放射性标记的适体用于 非人类灵长类动物的全身PET。初步结果表明，适体可以快速结合 凝血酶在体内对活性血栓的结合，并且解毒剂可以在5分钟内逆转这种结合。在 具体目标1，我们将通过在小鼠中使用适体-解毒剂进行研究来进一步开发该技术 对凝血酶、血管性血友病因子和血小板蛋白GPIIb/IIIa单独和组合产生。 在特定目标2中，将研究结果改编为使用全身的翻译恒河猴模型系统。 PET显示出出色的灵敏度。这些调查提出了一项新的战略，以解决 血栓在体内的表征，包括一个多学科的翻译团队与专业知识的核酸 酸生物化学、组合化学、抗血栓剂、新型诊断成像工具和 方法，以及具有深远翻译重要性的非人类灵长类动物模型系统。