Genetic and therapeutic studies of hemostatic and thrombotic disorders using zebrafish

使用斑马鱼进行止血和血栓性疾病的遗传和治疗研究

• 批准号: 9894256
• 负责人: JORDAN A SHAVIT
• 金额: $ 78万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2027-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9894256
• 关键词: Affect; Anticoagulants; Anticoagulation; Biological; Biological Assay; Blood Coagulation Disorders; Blood Coagulation Factor; Chemicals; Clustered Regularly Interspaced Short Palindromic Repeats; Coagulation Process; Diagnostic; Disease; Family; Fishes; General Population; Genes; Genetic; Genetic study; Genome; Health; Hemorrhage; Hemorrhagic Disorders; Hemostatic Disorders; Hemostatic function; Human; Individual; Intravenous infusion procedures; Lead; Life; Light; Mutagenesis; Oral; Pathologic; Pathology; Patients; Penetrance; Protein Engineering; Recombinant Proteins; Recombinants; Regulator Genes; Severities; Severity of illness; System; Therapeutic; Therapeutic Studies; Thrombopoiesis; Thrombosis; Work; Zebrafish; blood product; experimental study; genetic technology; genome editing; high risk; human morbidity; human mortality; individualized medicine; innovation; mutant; new therapeutic target; next generation sequencing; novel; novel diagnostics; novel therapeutics; nuclease; prospective; public health relevance; therapeutic target; tool; transcription activator-like effector nucleases

PROJECT SUMMARY/ABSTRACT Pathologic dysregulation of the coagulation system is a major contributor to human morbidity and mortality, resulting in either excessive bleeding or clotting. Significant progress has been made in the identification of genetic regulators of the coagulation cascade, but many unknown modifier genes contribute to the variable disease severity and penetrance observed among patients and families with hemostatic and thrombotic disorders. Understanding such modifiers could help classify patients at higher risk for pathology as well as identify novel therapeutic targets. Although major improvements to treatment of hemorrhagic and bleeding disorders have been made with blood products and human derived or recombinant coagulation factors, these have limited shelf life and storage conditions, and require intravenous infusion. Anticoagulation has seen a surge in recent years with many new direct acting oral anticoagulants, but their mechanisms of action are limited to the coagulation cascade. Building on our previous work, this project will take advantage of powerful genetic tools, including genome editing nucleases, next generation sequencing, and the zebrafish. We will conduct a large scale interrogation of the genome to discover hemostasis regulatory genes with the potential to modify the severity of human coagulation disorders. We have developed a panel of clotting factor mutant zebrafish using robust genome editing nucleases (TALENS and CRISPR/Cas) and conducted chemical mutagenesis experiments that have identified potential suppressor mutant lines harboring prospective thrombosis and hemostasis modifier genes. This panel of mutant fish will also be used for unbiased assays to identify novel lead molecules that suppress hemorrhage or thrombosis. The approaches described in this proposal will lead to the identification of the key non-canonical factors regulating hemostasis and thrombopoiesis, some of which will likely prove to be important genetic modifying factors in humans, or will suggest novel species-specific but biologically insightful regulatory mechanisms. This will shed light on the regulatory mechanisms of hemostasis, and the modifiers will also be candidate diagnostic and therapeutic targets for human thrombotic and hemorrhagic diseases. These targets will be utilized to develop potential innovative agents and new therapeutic classes for treatment of hemorrhage and thrombosis that could benefit the general population, as well as patients with bleeding and thrombotic disorders.

项目摘要/摘要 凝血系统的病理失调是人类发病率和 死亡率，导致过度出血或凝结。在 识别凝血级联的遗传调节剂，但许多未知的修饰基因有助于 疾病的严重程度和止血性患者和家庭中观察到的可变疾病严重程度和外观 血栓性疾病。了解此类修饰符可以帮助将病理风险较高的患者分类为 以及确定新颖的治疗靶标。尽管对出血和治疗的重大改善 血液产品和人类衍生或重组凝结已经出血疾病 因素，这些保质期有限和储存条件，需要静脉输注。抗凝 近年来，有许多新的直接表演口服抗凝剂激增，但它们的机制 动作仅限于凝血级联。在我们以前的工作的基础上，该项目将利用 强大的遗传工具，包括基因组编辑核酸酶，下一代测序和斑马鱼。 我们将对基因组进行大规模的询问，以发现与 改变人类凝血疾病的严重程度的潜力。我们已经开发了一个凝血因子的面板 使用鲁棒基因组编辑核酸酶（Talens和CRISPR/CAS）的突变斑马鱼并进行了化学 已经确定了具有前瞻性的潜在抑制突变线的诱变实验 血栓形成和止血改性基因。该突变鱼面板也将用于公正的测定法 确定抑制出血或血栓形成的新型铅分子。其中描述的方法 提案将导致确定调节止血和调节的关键非规范因素 血栓形成，其中一些可能被证明是人类中重要的遗传修饰因素，或者将是 建议新的物种特异性但具有生物学上有见地的调节机制。这将使 止血和修饰符的调节机制也将是候选诊断和治疗 人类血栓形成和出血性疾病的靶标。这些目标将用于开发潜力 创新的药物和新的治疗出血和血栓形成的治疗类别，可能受益 普通人群以及出血和血栓性疾病的患者。