A novel thrombolytic targeting Von Willebrand Factor (VWF) to treat ischemic stroke

一种针对血管性血友病因子 (VWF) 治疗缺血性中风的新型溶栓药物

• 批准号: 10289825
• 负责人: Shahid Nimjee
• 金额: $ 56.4万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-15 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10289825
• 关键词: Acute; Adhesions; Affect; Affinity; Age; Age Factors; Alteplase; Animal Model; Architecture; Area; Autologous; Binding; Biological Specimen database; Blood; Blood - brain barrier anatomy; Blood Platelets; Blood coagulation; Blood specimen; Caring; Carotid Artery Thrombosis; Cell Adhesion; Cerebral Edema; Cerebral Thrombus; Cerebrum; Cessation of life; Clinical; Coagulation Process; Data; Encapsulated; Endothelium; Erythrocytes; Event; Factor VIII; Fibrin; Fibrinolytic Agents; Genetic; Glycoprotein Ib; Glycoproteins; Goals; Hemorrhage; Hour; Hypertension; In Vitro; Infarction; Institution; Intracranial Hemorrhages; Ischemic Stroke; Laser Speckle Imaging; Ligands; Magnetic Resonance Imaging; Measures; Mechanics; Mediating; Mediator of activation protein; Morbidity - disease rate; Mus; Oligonucleotides; Outcome; Patients; Pharmaceutical Preparations; Plasma; Platelet Activation; Platelet Glycoprotein GPIb-IX Complex; Platelet aggregation; Play; Pre-Clinical Model; Process; Production; Proteins; RNA; Recovery; Recovery of Function; Resistance; Risk; Role; Safety; Severities; Specificity; Stroke; Testing; Thrombectomy; Thrombin; Thromboembolism; Thrombosis; Thrombus; Time; Translating; Translational Research; Work; aged; aptamer; artery occlusion; base; behavior test; clinical database; design; disability; effective therapy; endovascular thrombectomy; functional outcomes; improved; improved outcome; in vivo; injured; innovation; mortality; mouse model; novel; oligomycin sensitivity-conferring protein; post stroke; prevent; programs; receptor; restenosis; scaffold; sex; stroke incidence; stroke intervention; stroke model; stroke patient; stroke symptom; stroke therapy; thromboembolic stroke; thrombolysis; von Willebrand Factor

PROJECT SUMMARY Arterial thrombosis resulting in acute ischemic stroke (AIS) is the leading cause of combined morbidity and mortality worldwide. Recombinant tissue plasminogen activator (rtPA) is the only drug approved to treat AIS. Unfortunately, only ~5-10% of patients who present with AIS actually receive rtPA. Risks of rtPA treatment include a significant increase in symptomatic intracranial hemorrhage (ICH), which occurs in up to 6.4% of patients who receive the drug. Moreover, rtPA only achieves 10% recanalization in patients who present with large vessel occlusion (LVO) stroke. These clots, which are commonly platelet-rich, are notoriously resistant to rtPA. A critical need exists to develop thrombolytic agents that: 1. target critical proteins involved in stroke clot architecture, 2. recanalize arterial occlusions, and 3. have a safety profile superior to rtPA. Von Willebrand Factor (VWF) is an optimal target for AIS treatment. VWF binds to glycoprotein Ib (GPIb) of the platelet receptor complex GPIb-IX-V as well as to GPIIb-IIIa, resulting in platelet activation and aggregation. VWF also self-associates, extending into the vessel lumen as a scaffold for platelet and red blood cell adhesion. These processes result in arterial thrombosis as seen in AIS patients. Our preliminary data of cerebral thrombi from stroke patients show that the majority of clots have a platelet shell rich with VWF encapsulating the thrombus core, providing an explanation for the poor arterial recanalization rate associated with rtPA. Aptamers are oligonucleotide-based drugs that inhibit their target proteins with high affinity and specificity. We have isolated and optimized an RNA aptamer that binds to and inhibits VWF (DTRI-031). We have also designed a second oligonucleotide (DTRI-025) that fully reverses DTRI-031 activity within minutes. Our data in small and large animal models of thrombosis demonstrates that DTRI-031 both prevents thrombus formation and lyses fully formed arterial occlusions better than rtPA. The overall goals of this proposal are to 1) correlate elevated plasma VWF to clot VWF in AIS patients and 2) demonstrate VWF inhibition by DTRI-031 can translate into an effective treatment for patients who present with AIS. We will test the hypotheses that 1) VWF is an optimal target for AIS treatment. Our preliminary data shows that LVO AIS patients have significantly elevated plasma VWF. Our preliminary data has replicated these findings in a murine model of stroke. 2) DTRI-031 effectively lyses clots in vitro in blood samples from AIS patients and in vivo in a murine model of stroke. Moreover, DTRI-025 rapidly reverses DTRI-031 activity in AIS in vitro and in vivo. 3) DTRI-031 treatment improves outcomes in a murine model of AIS by increasing recanalization, decreasing infarct volume, and improving functional recovery. Our preliminary data reveals that DTRI-031-treated mice have reduced post-stroke infarct volumes compared to control. Finally, DTRI-031 has an improved safety profile by decreasing ICH, cerebral edema formation and blood-brain-barrier breakdown after AIS compared to rtPA.

项目概要 动脉血栓形成导致急性缺血性中风（AIS）是发病率和死亡率的主要原因 全球死亡率。重组组织纤溶酶原激活剂 (rtPA) 是唯一被批准用于治疗 AIS 的药物。 不幸的是，只有约 5-10% 的 AIS 患者实际上接受了 rtPA。 rtPA 治疗的风险 包括症状性颅内出血 (ICH) 的显着增加，这种情况的发生率高达 6.4% 接受该药物的患者。此外，对于出现以下症状的患者，rtPA 只能实现 10% 的再通 大血管闭塞（LVO）中风。这些凝块通常富含血小板，众所周知，它们具有抵抗力 rtPA。迫切需要开发能够： 1. 靶向参与的关键蛋白质 中风血栓结构，2. 使动脉闭塞再通，3. 安全性优于 rtPA。 血管性血友病因子 (VWF) 是 AIS 治疗的最佳目标。 VWF 与糖蛋白 Ib (GPIb) 结合 血小板受体复合物 GPIb-IX-V 以及 GPIIb-IIIa，导致血小板活化和聚集。 VWF 还可以自我结合，延伸到血管腔中作为血小板和红细胞粘附的支架。 这些过程会导致动脉血栓形成，如 AIS 患者所见。我们的脑血栓初步数据 来自中风患者的研究表明，大多数凝块具有富含 VWF 的血小板壳，包裹着 血栓核心，为与 rtPA 相关的动脉再通率低下提供了解释。 适体是基于寡核苷酸的药物，能够以高亲和力和特异性抑制其靶蛋白。我们 分离并优化了一种可结合并抑制 VWF (DTRI-031) 的 RNA 适体。我们还有 设计了第二种寡核苷酸（DTRI-025），可在几分钟内完全逆转 DTRI-031 活性。我们的 小型和大型动物血栓形成模型的数据表明，DTRI-031 可以预防血栓 比 rtPA 更好地形成和溶解完全形成的动脉闭塞。该提案的总体目标是 1) 将 AIS 患者升高的血浆 VWF 与凝块 VWF 关联起来，2) 证明 DTRI-031 抑制 VWF 可以 转化为对 AIS 患者的有效治疗。 我们将检验以下假设：1) VWF 是 AIS 治疗的最佳目标。我们的初步数据显示 LVO AIS 患者的血浆 VWF 显着升高。我们的初步数据复制了这些 在小鼠中风模型中的发现。 2) DTRI-031 可在体外有效溶解 AIS 血样中的凝块 患者和中风小鼠模型的体内。此外，DTRI-025 快速逆转 AIS 中的 DTRI-031 活性 体外和体内。 3) DTRI-031 治疗通过增加 AIS 小鼠模型的治疗效果来改善结果 再通，减少梗塞体积，改善功能恢复。我们的初步数据表明 与对照组相比，DTRI-031 治疗的小鼠中风后梗塞体积减少。最后，DTRI-031 有一个 通过减少脑出血、脑水肿形成和血脑屏障破坏来提高安全性 AIS 与 rtPA 的比较。