Understanding and Controlling the Contribution of Fibrinolysis to Bleeding Using a Long-Acting Antifibrinolytic RNA Therapy

使用长效抗纤溶 RNA 疗法了解和控制纤溶对出血的影响

• 批准号: 10737327
• 负责人: Christian Kastrup
• 金额: $ 75.73万
• 依托单位: VERSITI WISCONSIN, INC.
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-15 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10737327
• 关键词: Acute; American; Animal Model; Animals; Antifibrinolytic Agents; Antiplasmin; Biochemistry; Bleeding time procedure; Blood Coagulation Disorders; Canis familiaris; Clinical; Coagulation Process; Data; Development; Diagnosis; Disease; Disease model; Dose; Enzyme Precursors; Enzymes; Equilibrium; FDA approved; Family suidae; Fibrinolysis; Fibrinolytic Agents; Gene Silencing; Genetic; Half-Life; Health; Hemophilia A; Hemophilia B; Hemorrhage; Hemostatic Agents; Hemostatic function; Hour; In Vitro; Incidence; Inherited; Injury; Investigation; Knowledge; Measures; Mediating; Menorrhagia; Methods; Modeling; Morbidity - disease rate; Mus; Nerve; Operative Surgical Procedures; Outcome; Patients; Persons; Plasmin; Plasminogen; Plasminogen Activator Inhibitor 1; Predisposition; Process; Prophylactic treatment; Proteins; Puncture procedure; Quality of life; RNA; RNA-targeting therapy; Research; Research Personnel; Role; Rupture; Safety; Saphenous Vein; Severities; Small Interfering RNA; Symptoms; Tail; Testing; Therapeutic; Toxic effect; Tranexamic Acid; Trauma; Traumatic Hemorrhage; Traumatic injury; Woman; Work; canine model; comparison control; gene therapy; improved; in silico; in vivo; innovation; insight; joint injury; knock-down; lipid nanoparticle; mortality; mouse model; novel; porcine model; prevent; prophylactic; small molecule inhibitor; targeted agent; targeted treatment; therapeutic RNA; therapeutic nanoparticles; therapy development; tool; von Willebrand Disease

SUMMARY BACKGROUND: The balance between clot formation (coagulation) and degradation (fibrinolysis) is disrupted in patients with bleeding disorders or severe hemorrhage, resulting in increased bleeding. The extent to which fibrinolysis contributes to bleeding from the onset of acute bleeding in these contexts is unclear, and this gap in knowledge limits the development of therapies that inhibit fibrinolysis. Current therapeutics are often not sufficient for patients with bleeding disorders; for example, antifibrinolytic drugs are widely used to manage acute bleeding in these patients, but are unsuitable for long-term prophylaxis due to short half-lives that decrease efficacy. RATIONALE: We hypothesize that fibrinolysis contributes to the incidence and severity of bleeding starting from the onset of bleeding, and that long-acting RNA agents targeting plasminogen, the zymogen precursor of the main fibrinolytic enzyme plasmin, can decrease fibrinolysis and help manage bleeding long-term for people across all classes of bleeding disorders, including diagnosed and undiagnosed disorders. This knowledge will provide insights into the mechanisms that impact outcomes in traumatic hemorrhage. These RNA agents will be delivered using lipid nanoparticles (LNPs), a clinically approved delivery platform we have unique expertise in. SPECIFIC AIMS: We will develop an RNA therapeutic targeting plasminogen, using small interfering RNA (siRNA)-mediated gene silencing (siPlg). This therapy is long-acting over several weeks per dose and highly specific for the target protein, which decreases the burden of frequent administration. The specific aims of this proposal are to (1) Develop approaches for knockdown of plasminogen and inhibition of fibrinolysis in mice, pigs, and dogs, and characterize the safety of this approach; (2) Determine the contribution of fibrinolysis to blood loss at the onset and in early stages of hemorrhage in a traumatic injury model in swine; and (3) Investigate the role of fibrinolysis in bleeding disorders by testing the effect of plasminogen knockdown in mouse and canine models of hemophilia A, and mouse models of hemophilia B and von Willebrand disease. INNOVATION: This work has both mechanistic and therapeutic innovation. It will immediately determine the contribution of fibrinolysis to bleeding in early stages of severe hemorrhage and in bleeding disorders. This will likely be the first development of RNA-LNP therapies targeting circulating proteins in large animal models of bleeding. These findings will address a fundamental question in trauma and hemostasis research. Separately, it will aid the development of improved, long-acting therapies for controlling bleeding in bleeding disorders. These therapies would be particularly beneficial for women with menorrhagia caused by bleeding disorders. EXPECTED OUTCOMES: We expect to (1) Develop species-specific siPlg that have minimal adverse health effects; and (2-3) Demonstrate that knocking down plasminogen inhibits fibrinolysis and decreases blood loss in swine models of traumatic hemorrhage, and in murine and canine models of bleeding disorders.

概括 背景：血栓形成（凝血）和降解（纤维蛋白溶解）之间的平衡被破坏 患有出血性疾病或严重出血，导致出血增多的患者。程度 在这些情况下，纤维蛋白溶解是否会导致急性出血发作时的出血尚不清楚，并且这种差距 知识限制了抑制纤维蛋白溶解疗法的发展。目前的治疗方法往往不够 对于患有出血性疾病的患者；例如，抗纤溶药物广泛用于治疗急性出血 但由于半衰期短会降低疗效，因此不适合长期预防。 理由：我们假设纤维蛋白溶解会导致出血的发生率和严重程度 出血的发生，以及针对纤溶酶原（纤溶酶原的酶原前体）的长效 RNA 药物 主要纤维蛋白溶解酶纤溶酶，可以减少纤维蛋白溶解并帮助人们长期控制出血 涵盖所有类别的出血性疾病，包括已诊断和未诊断的疾病。这些知识将 提供对影响创伤性出血结果的机制的见解。这些 RNA 试剂将 使用脂质纳米颗粒 (LNP) 进行递送，这是一种经过临床批准的递送平台，我们拥有独特的专业知识。 具体目标：我们将使用小干扰 RNA 开发一种针对纤溶酶原的 RNA 治疗方法 (siRNA) 介导的基因沉默 (siPlg)。该疗法每剂可在数周内发挥长效作用，且疗效高 特异性针对靶蛋白，减轻频繁给药的负担。本次活动的具体目标 建议 (1) 开发在小鼠、猪、 和狗，并描述这种方法的安全性； (2)测定纤溶对血液的贡献 猪创伤性损伤模型中出血开始时和早期阶段的损失； (3) 调查 通过测试小鼠和犬中纤溶酶原敲低的效果来研究纤维蛋白溶解在出血性疾病中的作用 血友病 A 模型，以及血友病 B 和冯维勒布兰德病的小鼠模型。 创新：这项工作在机制和治疗上都有创新。它将立即确定 纤维蛋白溶解对严重出血和出血性疾病早期出血的贡献。这将 可能是针对大型动物模型中循环蛋白的 RNA-LNP 疗法的首次开发 流血。这些发现将解决创伤和止血研究中的一个基本问题。分别地，它 将有助于开发改进的长效疗法来控制出血性疾病的出血。这些 治疗对于因出血性疾病引起月经过多的女性特别有益。 预期成果：我们期望 (1) 开发对健康影响最小的物种特异性 siPlg 影响； (2-3) 证明敲除纤溶酶原可抑制纤维蛋白溶解并减少失血 猪的创伤性出血模型，以及鼠和犬的出血性疾病模型。