Blocking the immune response to HDAd for Hemophilia A gene therapy

项目名称。 重组血友病A基因辅助型腺病毒介导的免疫应答阻断 疗法 项目摘要。 血友病A（Hemophilia A，HA）是一种常见的凝血功能障碍性疾病，由凝血因子VIII（Factor VIII，FVIII）缺乏引起。 主要的治疗方法是使用重组人FVIII进行替代治疗。然而，在这方面， 由于高成本、治疗的可及性和抑制性抗体的形成， 长期的发病率和死亡率。我们开发了一种优化的依赖于助手的 腺病毒基因载体（HDAds）系统使我们能够实现两种分泌的 和无慢性毒性和持久性的细胞内转基因的载体使用小和大 动物模型然而，急性毒性仍然是临床转化的障碍。为了克服这一点，我们 提出开发表达SOCS 1和/或编码TLR 9抑制剂的免疫抑制性HDAs 序列的然而，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连续性治疗，该治疗已经具有临床益处， 在败血症的情况下去除急性免疫成分。我们将联合收割机血液滤过与 在非人灵长类动物的安全性研究中，改善了免疫抑制HDAs。最终，我们将应用 该方法用于临床前治疗鼠和犬HA中的FVIII缺陷。解决 潜在的适应性免疫应答FVIII治疗，我们将共表达血管性血友病因子（vWF） 关于FVIII通过这些研究，我们将解决基因治疗中的三个重要问题， i）我们能降低对HDAs的先天免疫反应吗？（二）能否改善 HA模型中FVIII表达的有效性？iii）我们能降低先天免疫反应吗？ 非人灵长类动物模型进行连续性血液滤过？此应用程序的总体目标是 建立安全有效的辅助依赖性腺病毒载体HA基因治疗 结合血液滤过，其可以容易地在临床竞技场中转化以用于未来的临床试验。 在此指导阶段（K99），我将通过以下方式提高HA治疗的HDAd治疗指数： 结合先天性免疫应答的细胞自主免疫抑制， 非细胞自主性体液因子和FVIII稳定因子。辅导阶段（K99）将 发生在贝勒医学院的指导下，博士布伦丹李。在随后的 独立研究者阶段（R 00），我将把这种混合HDAs应用于犬HA，为 临床研究。我还将评价血液滤过辅助混合HDAs的治疗效果 注射到非人类灵长类动物身上

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.