Direct Detection and Characterization of Thrombosis In Vivo

项目摘要/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， 是手术后的重要并发症。目前用于评估血栓事件的成像方式 提出挑战，因为它们主要是可视化的血液流动不足，而不是详细的信息 他们不能区分新形成的血栓和陈旧的血栓。我们的团队 已经证明，高亲和力和高特异性的RNA适配子可以针对 凝血酶和凝血因子IXa等凝血蛋白的数量。此外，我们的团队已经证明了 适体-解毒剂对可作为快速结合-快速逆转探针用于实时检测血栓。 本申请中提出的研究将适配子-解毒剂对与灵敏的全身正电子相结合 发射断层扫描(PET)成像开发了一种治疗血栓事件患者的新方法 可以分层，然后进行治疗。这一提议代表了多学科的融合 专业领域，探索新的团队方向，将通过 具体目标如下：(1)开发和评价适体-解毒剂对，以执行血栓的快速成像 体内分子血栓的小鼠模型，以及(2)评估放射性标记适体对 非人灵长类的全身正电子发射计算机断层扫描初步结果表明，适配子可以快速结合 凝血酶在活体血栓上的结合，解毒剂可以在5分钟内逆转这种结合。在……里面 具体目标1，我们将通过在小鼠身上进行适体解毒剂的研究来进一步发展这项技术 凝血酶、血管性血友病因子和血小板蛋白GPIIb/IIIa单独和组合产生的对。 研究结果将在特定目标2中适用于使用全身的平移恒河猴模型系统。 表现出非凡敏感度的宠物。这些调查提出了一项新的战略来解决 体内血栓的特征，并包括一个具有核专业知识的多学科翻译团队 酸性生物化学、组合化学、抗血栓药物、新型诊断成像工具和 方法，以及一个具有深远翻译重要性的非人类灵长类动物模型系统。

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.