Biomarkers for 12-lipoxygenase inhibition as a therapeutic intervention for heparin-induced thrombocytopenia and thrombosis (HIT/T)

12-脂氧合酶抑制的生物标志物作为肝素诱导的血小板减少症和血栓形成的治疗干预措施 (HIT/T)

• 批准号: 10177358
• 负责人: MICHAEL Allan HOLINSTAT
• 金额: $ 20.86万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-11 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10177358
• 关键词: 12-HETE; Address; Amputation; Animal Model; Antibodies; Anticoagulants; Antithrombins; Arachidonate 12-Lipoxygenase; Argatroban; Autoantibodies; Bioavailable; Biological Assay; Biological Markers; Blood; Blood Platelets; Blood coagulation; Blood specimen; Cessation of life; Clinic; Clinical; Clinical Treatment; Clinical Trials; Coagulation Process; Complex; Deep Vein Thrombosis; Development; Diagnosis; Disease; Effectiveness; Environment; Enzyme-Linked Immunosorbent Assay; Enzymes; Exposure to; Fibrin; Gangrene; Generations; Goals; Grant; Hematological Disease; Hemorrhage; Heparin; Human; Hydroxyeicosatetraenoic Acids; Immune response; Injury; LOX gene; Lasers; Lead; Life; Limb structure; Low-Molecular-Weight Heparin; Mesentery; Metabolic; Morbidity - disease rate; Mus; PF4 Gene; Pathologic; Pathology; Patients; Platelet Activation; Platelet Count measurement; Platelet aggregation; Production; Publishing; Pulmonary Embolism; Rare Diseases; Reporting; Reproducibility; Risk; Syndrome; Testing; Therapeutic; Therapeutic Intervention; Thrombin; Thrombosis; Thrombus; Treatment Efficacy; Venous; Venous Thrombosis; Whole Blood; arteriole; base; clinical trial readiness; disease diagnosis; electric impedance; heparin-induced thrombocytopenia; immunothrombosis; in vivo; inhibitor/antagonist; light transmission; limb ischemia; mortality; mouse model; patient response; prevent; receptor; thrombotic complications

The goal of this project is to develop highly sensitive and reproducible biomarkers for assessment of VLX-1005 treatment of heparin-induced thrombocytopenia and thrombosis (HIT/T). VLX-1005, a 12-LOX inhibitor that potently inhibits immune-mediated thrombosis including HIT/T, has the potential to treat HIT/T in patients. HIT/T is an acquired life-threatening thrombocytopenic and thrombotic complications that occur in patients exposed to unfractionated heparin (UFH) or low molecular weight heparin (LMWH): the most widely used anticoagulants in the world. Approximately 12 million patients are exposed to UFH and LMWH annually in the US alone with ~24,000 developing HIT/T every year. HIT/T often results in aberrant platelet activation and aggregation leading to a) platelet-derived microparticle release, b) increased thrombin generation leading to catastrophic arterial and venous thrombosis that results in venous limb gangrene, c) deep venous thrombosis, and d) pulmonary embolism and death. Current treatment of HIT/T is limited to argatroban (a direct thrombin inhibitor). Unfortunately, this carries a significant risk of severe bleeding limited its effectiveness. Recent evidence has established that the enzyme 12- lipoxygenase (12-LOX), and its metabolic product, 12- hydroxyeicosatetraenoic acid (12-HETE) are key contributors to the underlying pathology of HIT/T including promoting platelet reactivity, thrombus formation/stability as well as vessel occlusion. VLX-1005 selectively and potently inhibits 12-LOX, halts aberrant platelet activation and thrombosis and has shown efficacy in numerous animal models, including mesenteric arteriole injury and laser-induced cremaster arteriole thrombosis. While the development of VLX-1005 for treatment of HIT/T is rapidly progressing, it will be important to have highly sensitive biomarkers for assessment of VLX-1005 as it intervenes and prevents morbidity and mortality often observed with the disease. To this end, we propose 2 Aims to address this concern for clinical trial readiness. In Aim 1, several biomarkers for coagulation state and platelet activation will be assessed including the thrombin-antithrombin (TAT) ELISA assay, 12-HETE assay, light transmission aggregometry (LTA), and whole blood impedance aggregometry (WBIA) in mouse models of HIT/T. These biomarkers will serve as an indicator of VLX-1005 effects on both coagulation and platelet activation. As VLX-1005 has not been shown to effect coagulation or fibrin formation, we do not expect VLX-1005 to alter coagulation conditions in the patient, however since a decreased platelet activation environment may result in decreased thrombin activation, these biomarker will inform if VLX-1005 is altering upstream coagulation. Aim 2 will take the biomarker optimization from Aim 1 and apply it to human blood samples ex vivo to determine the efficacy of the biomarkers to report VLX-1005 effects in the blood. The biomarker assay profile is essential for the planned clinical trials with VLX- 1005 for treatment of patients with HIT/T.

该项目的目的是开发高度敏感和可重复的生物标志物来评估VLX-1005 治疗肝素诱导的血小板减少症和血栓形成（HIT/T）。 VLX-1005，一种12-lox抑制剂 有效抑制免疫介导的血栓形成在内，包括hit/t，有可能治疗患者的命中率。 HIT/T是患者发生的一种危及生命的血小板减少和血栓形成并发症 暴露于未分离的肝素（UFH）或低分子量肝素（LMWH）：使用最广泛的 世界上的抗凝剂。每年约有1200万患者暴露于UFH和LMWH 我们一个人每年都有约24,000次命中率。命中/t通常会导致异常血小板激活和 聚集导致a）血小板衍生的微粒释放，b）凝血酶产生增加导致 灾难性动脉和静脉血栓形成导致静脉四肢坏疽，c）深静脉血栓形成， d）肺栓塞和死亡。命中/T的当前治疗仅限于Argatroban（直接凝血酶 抑制剂）。不幸的是，这有重大出血的重大风险限制了其有效性。最近的 有证据表明，酶12-脂氧酶（12-lox）及其代谢产物12- 羟基乙酸烯酸（12-HETE）是造成命中/T的潜在病理的关键因素 促进血小板反应性，血栓形成/稳定性以及血管阻塞。 VLX-1005有选择地 有效抑制12-lox，停止异常血小板激活和血栓形成，并显示出众多功效 动物模型，包括肠系膜动脉损伤和激光诱导的Cremaster动脉血栓形成。尽管 VLX-1005用于治疗命中/T的VLX-1005的发展迅速发展，具有高度的重要性 敏感的生物标志物用于评估VLX-1005，因为它介入并防止发病率和死亡率 观察到这种疾病。为此，我们提出2个旨在解决临床试验准备就绪的关注。 在AIM 1中，将评估几种用于凝血状态和血小板激活的生物标志物 凝血酶 - 抗凝血酶（TAT）ELISA分析，12-HETE分析，光透射聚集（LTA）和整个 HIT/T小鼠模型中的血液阻抗聚集法（WBIA）。这些生物标志物将作为指标 VLX-1005对凝结和血小板激活的影响。由于尚未证明VLX-1005的作用 凝血或纤维蛋白的形成，我们预计VLX-1005不会改变患者的凝血条件， 但是，由于血小板激活环境降低可能导致凝血酶激活降低，因此 生物标志物将告知VLX-1005是否正在改变上游凝血。 AIM 2将进行生物标志物优化 从AIM 1使用，并将其应用于人体样本，以确定生物标志物报告的功效 VLX-1005在血液中的影响。生物标志物测定概况对于计划的VLX-计划进行的临床试验至关重要 1005用于命中/T患者的治疗。