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.

项目摘要 导致急性缺血性卒中（AIS）的动脉血栓形成是发病率组合的主要原因 全球死亡率。重组组织纤溶酶原激活剂（RTPA）是唯一批准治疗AIS的药物。 不幸的是，有AIS的患者中，只有约5-10％的人实际上接受了RTPA。 RTPA治疗的风险 包括症状性颅内出血（ICH）的显着增加，最多发生在6.4％ 接受该药物的患者。此外，RTPA仅在出现的患者中实现10％ 大容器阻塞（LVO）中风。这些通常富含血小板的凝块，众所周知 RTPA。存在关键需求，以开发溶栓剂：1。靶向涉及的关键蛋白 中风凝块结构，2。重新构成动脉闭塞，3。具有比RTPA优于RTPA的安全性。 von Willebrand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，导致血小板激活和聚集。 VWF还自缔合，延伸到血管腔中，作为血小板和红细胞粘合剂的支架。 这些过程导致AIS患者所见的伪影。我们的大脑血栓的初步数据 从中风患者表明，大多数血块具有富含VWF的血小板外壳，可封装 血栓核心，为与RTPA相关的动脉再通率较差提供了解释。 APATMER是基于寡核苷酸的药物，可抑制其高亲和力和特异性的靶蛋白。我们 已经分离并优化了与VWF结合并抑制VWF的RNA APATMER（DTRI-031）。我们也有 设计了第二个寡核苷酸（DTRI-025），该寡核苷酸（DTRI-025）在几分钟内完全逆转了DTRI-031活动。我们的 大小动物模型的数据表明，DTRI-031都可以防止血栓 形成和裂解比RTPA更好地形成动脉闭塞。该提议的总体目标是1） 在AIS患者中，将升高的血浆VWF与凝块VWF相关，2）证明了DTRI-031抑制VWF CAN 转化为患有AIS的患者的有效治疗方法。 我们将测试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。我们的初步数据已复制这些 鼠类中风模型中的发现。 2）DTRI-031在AIS的血液样本中有效地在体外裂解布 患者和体内的鼠类中风模型。此外，DTRI-025迅速逆转AIS中的DTRI-031活性 体外和体内。 3）DTRI-031治疗通过增加AIS的鼠模型中的结果 重新定性，减少梗塞量并改善功能恢复。我们的初步数据表明 与对照组相比，经DTRI-031处理的小鼠减少了冲程后梗塞体积。最后，DTRI-031有一个 通过减少ICH，脑水肿的形成和血脑屏障分解，改善了安全性。 与RTPA相比，AIS。