Optimization of AAV vector to deliver FVa for hemophilia with inhibitors

优化 AAV 载体以通过抑制剂递送血友病 FVa

• 批准号: 10372097
• 负责人: Chengwen Li
• 金额: $ 62.56万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-03-01 至 2024-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10372097
• 关键词: Animal Model; Antibodies; Blood; Blood Coagulation Disorders; Blood Coagulation Factor; Bypass; Canis familiaris; Clinical; Clinical Trials; Coagulation Factor Deficiency; Coagulation Process; Complementary DNA; Complication; Dependovirus; Development; Dose; F8 gene; Factor IX; Factor V; Factor VIII; Factor Va; Factor X; Financial Hardship; Future; Gene Delivery; Gene Transfer; Generations; Genome; Hemophilia A; Hemophilia B; Hemorrhage; Hemostatic function; Hepatocyte; Hospitalization; Infusion procedures; Injections; Introns; Light; Link; Liver; Mediating; Modification; Mus; Pathway interactions; Patients; Phenotype; Proteins; Prothrombin; Replacement Therapy; Single-Stranded DNA; Therapeutic Effect; Thrombin; Thromboplastin; Transgenes; Treatment Cost; Virion; Wild Type Mouse; adeno-associated viral vector; arthropathies; base; design; experimental study; gene product; gene therapy; improved; inhibitor; pre-clinical; prevent; promoter; transgene expression; treatment strategy; vector

ABSTRACT Gene therapy with adeno-associated virus (AAV) vectors have been successfully applied in hemophilia patients. However, the patients with inhibitors (antibodies against coagulation factors) are excluded from these trials. AAV vectors have also been explored for delivery of a bypass product, FVIIa transgene, in preclinical animal models. Although long-term improvement of hemostasis was achieved, the complete phenotypic correction was not observed. During the coagulation cascade, FV (FVa) functions as a co-factor of FXa to amplify thrombin generation. We pioneered a study in which FVa driven by a liver specific promoter was constructed and packaged into AAV virions. After injection of AAV/FVa vectors into hemophilic mice, completely phenotypic correction was achieved over 28 weeks without obvious complications. In this proposal, we will explore an effective strategy using AAV vectors to deliver bypass products in the treatment of hemophilia with inhibitors. First, we will optimize FVa constructs by utilization of different hepatocyte promoters and modification of linker sequences between the FV heavy chain and light chain (Aim 1). Next, we will explore whether the combination of AAV/FVa and AAV/FVIIa has a synergistic effect on the improvement of hemostasis in hemophilic settings (Aim 2). Since the results obtained in mice experiments often do not translate to large animal models, we propose to examine the long term phenotypic correction effect in hemophilic dogs using AAV/FVa vector alone or in combination with AAV/FVIIa (Aim 3). Overall, the studies proposed in the project will establish the basis for AAV vector-mediated bypass product gene transfer in future clinical trials in hemophilia patients with inhibitors. !

抽象的 腺相关病毒（AAV）载体的基因治疗已成功应用于血友病 患者。然而，患有抑制剂（抗凝血因子抗体）的患者被排除在外。 试验。在临床前研究中，AAV 载体也被探索用于传递旁路产物 FVIIa 转基因 动物模型。尽管实现了止血的长期改善，但完整的表型 没有观察到校正。在凝血级联过程中，FV (FVa) 作为 FXa 的辅助因子发挥作用， 增强凝血酶的产生。我们开创了一项研究，其中由肝脏特异性启动子驱动的 FVa 构建并包装成 AAV 病毒体。将AAV/FVa载体注射到血友病小鼠体内后， 28周内实现了完全的表型纠正，没有明显的并发症。在这个提案中， 我们将探索一种有效的策略，使用 AAV 载体来提供旁路产品来治疗 具有抑制剂的血友病。首先，我们将通过利用不同的肝细胞启动子来优化 FVa 构建体 以及 FV 重链和轻链之间的接头序列的修饰（目标 1）。接下来，我们将 探讨AAV/FVa与AAV/FVIIa联合使用是否对改善 血友病患者的止血（目标 2）。由于在小鼠实验中获得的结果通常并不 转化为大型动物模型，我们建议检查长期表型校正效应 单独使用 AAV/FVa 载体或与 AAV/FVIIa 联合使用的血友病犬（目标 3）。总体而言，研究 该项目提出的方案将为未来AAV载体介导的旁路产物基因转移奠定基础 在血友病患者中进行抑制剂的临床试验。 ！