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.

摘要 背景：血栓形成(凝血)和降解(纤溶)之间的平衡被破坏 患者出现出血紊乱或严重出血，导致出血增多。在多大程度上 在这些情况下，纤溶作用导致急性出血的原因尚不清楚，这一差距在 知识限制了抑制纤溶的治疗方法的发展。目前的治疗方法往往是不够的 对于有出血障碍的患者；例如，抗纤溶药物被广泛用于治疗急性出血。 在这些患者中，但由于半衰期短，降低了疗效，不适合长期预防。 理论基础：我们假设纤溶作用对出血的发生率和严重程度有贡献。 出血的开始，以及针对纤溶酶原的长效RNA制剂，纤溶酶原是 主要纤溶酶-纤溶酶，可减少纤溶，帮助人们长期止血 涵盖所有类别的出血性疾病，包括已诊断和未诊断的疾病。这一知识将会 深入了解影响创伤性出血预后的机制。这些核糖核酸试剂将是 使用脂质纳米粒(LNPs)给药，这是我们拥有独特专业知识的临床批准的给药平台。 具体目标：我们将开发一种针对纤溶酶原的RNA疗法，使用小干扰RNA (SiRNA)介导的基因沉默(SiPlg)。这种疗法的长效作用在每剂数周以上，而且 针对目标蛋白质，从而减少频繁给药的负担。这样做的具体目的是 建议是：(1)开发抑制小鼠、猪、 (2)测定纤溶作用对血液的贡献 在猪创伤模型中，失血起病和早期失血；以及(3)研究 纤溶酶原在小鼠和犬体内的作用及其在出血性疾病中的作用 血友病A模型，血友病B和血管性血友病小鼠模型。 创新：这项工作既有机制创新，也有治疗创新。它将立即确定 在严重出血的早期阶段和出血性疾病中，纤溶对出血的贡献。这将是 可能是针对循环蛋白的RNA-LNP疗法在大型动物模型中的第一次开发 在流血。这些发现将解决创伤和止血研究中的一个基本问题。另外，它还 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 对于因出血性疾病导致月经过多的妇女来说，治疗尤其有益。 预期结果：我们希望(1)开发出对健康危害最小的特定物种siplg 影响；和(2-3)证明击倒纤溶酶原抑制纤溶和减少失血。 猪的创伤性出血模型，以及小鼠和犬的出血性疾病模型。