Blocking the immune response to HDAd for Hemophilia A gene therapy

项目名称。 阻断携带改良血友病A基因的辅助性腺病毒载体的免疫应答 心理治疗。 项目摘要。 血友病A(HA)是由凝血因子VIII(FVIII)缺乏引起的一种常见的凝血功能障碍。 治疗的主流是用重组人FVIII进行替代治疗。然而， 由于昂贵的费用、获得治疗的机会以及抑制抗体的形成，患者继续遭受痛苦。 从重大的长期发病率和死亡率。我们开发了一种优化的帮手依赖型 腺病毒基因载体(HDAds)系统使我们能够实现两种分泌物的长期表达 和细胞内转基因无慢性毒性和持久性的载体使用小和大 动物模型。然而，急性毒性仍然是临床翻译的障碍。为了克服这一点，我们 建议开发表达SOCS1和/或编码TLR9抑制物的免疫抑制HDAD 序列。然而，由于先天免疫系统的巨大冗余，我们建议 开发血液滤过形式的辅助治疗，已在临床上有益于 清除脓毒症环境中的急性免疫成分。我们将结合血液滤过和 在非人类灵长类动物的安全性研究中改进的免疫抑制HDADS。最终，我们将申请 本研究旨在探讨FVIII缺乏症的临床前治疗方法。要解决这个问题 对FVIII治疗的潜在获得性免疫反应，我们将联合表达von Willebrand因子(VWF) 与FVIII一起。通过这些研究，我们将解决基因治疗中的三个重要问题 血友病A I)我们能降低对HDADS的先天免疫反应吗？二)我们能否改进 FVIII在HA模型中的表达效果？III)我们能降低先天免疫反应吗 辅助血液滤过的非人灵长类动物模型？此应用程序的总体目标是 应用辅助性腺病毒载体(HDAds)建立安全有效的HA基因治疗 与血液滤过相结合，可以很容易地翻译到临床舞台上，用于未来的临床试验。 在这个指导阶段(K99)，我将通过以下方式提高HA治疗的HDAd治疗指数 结合细胞自主免疫抑制的先天免疫反应，物理清除 非细胞自主体液因子和FVIII稳定因子。指导阶段(K99)将 在布伦丹·李博士的指导下在贝勒医学院进行。在随后的 独立研究阶段(R00)，我将把这种混合HDADS应用于犬HA，为 临床研究。我还将评估血液滤过辅助混合HDADS的治疗效果 注射给非人类灵长类动物。

Project Title. Blocking the immune response to Helper-dependent adenovirus vector for improved Hemophilia A gene therapy. Project Abstract. Hemophilia A (HA) is a common disorder of coagulation caused by deficiency of factor VIII (FVIII). The mainstay of treatment has been replacement therapy with recombinant human FVIII. However, because of high cost, access to therapy, and inhibitory antibody formation, patients continue to suffer from significant long-term morbidity and mortality. Our development of an optimized helper-dependent adenoviral gene vector (HDAds) system has enabled us to achieve long term expression of both secreted and intracellular transgenes without chronic toxicity and persistence of vector using both small and large animal models. However, acute toxicity remains an obstacle to clinical translation. To overcome this, we propose to develop immune suppressive HDAds expressing SOCS1 and/or coding TLR9 inhibitor sequences. However, because of the great redundancy of the innate immune system, we propose to develop adjunctive therapy in the form of hemofiltration that has already to be clinically-beneficial to remove the acute immune components in the context of sepsis. We will combine hemofiltration with improved immunosuppressive HDAds in safety studies in nonhuman primates. Ultimately, we will apply this approach to the preclinical treatment of FVIII deficiency in murine and canine HA . To address the potential adaptive immune response to FVIII therapy, we will co-express von Willebrand Factor (vWF) with FVIII. With these studies, we will address three important questions in genetic therapy for hemophilia A i) Can we decrease the innate immune response to HDAds? ii) Can we improve the efficacy of FVIII expression in HA model? iii) Can we decrease the innate immune response in nonhuman primate model with adjunctive hemofiltration? The overall goal of this application is to establish the safe and effective HA gene therapy with helper-dependent adenoviral vectors (HDAds) combined with hemofiltration that can be readily translated in the clinical arena for future clinical trials. During this mentored phase (K99), I will improve the therapeutic index of HDAd for HA therapy by combining cell autonomous immune suppression of the innate immune response, physical clearance of non-cell autonomous humoral factors, and stabilization factor of FVIII,. The mentored phase (K99) will occur at Baylor College of Medicine under the guidance of Dr. Brendan Lee. In the subsequent independent investigator phase (R00), I will apply this hybrid HDAds to canine HA in preparation for clinical studies. I will also evaluate the therapeutic effect of hemofiltration-assisted hybrid HDAds injection in nonhuman primates.