Immunopharmacology of FVIII Bioengineering and Gene Therapy

项目3：FVIII生物工程和基因治疗的免疫药理学 总结 针对人因子VIII的抑制性抗体的开发仍然是最重要的临床应用。 血友病A是与血友病A治疗相关的并发症，是基因治疗方法的关键障碍。 因此，提高了对与有前途的肝靶向治疗相关的免疫学和药理学的理解， 腺相关病毒（AAV）载体和造血干细胞（HSC）定向慢病毒载体（LV）基因 需要治疗策略。U 54申请项目3中提出的研究旨在推进 我们在这些基因疗法背景下对FVIII免疫生物学的了解。拟议的研究旨在 分析生物工程重组输注产品的分子组成和免疫反应性 以及肝脏定向的AAV和HSC定向的LV基因疗法衍生的fVIII，ii）鉴定关键设计 与肝定向AAV-fVIII基因的免疫原性和抑制剂放射潜力相关的参数 iii）评估非遗传毒性HSC定向的免疫原性和抑制剂根除潜力， LV基因治疗方案。这些研究将利用现有的FVIII、糖组学、结构生物学， 免疫学和基因治疗专业知识以及各种最先进的技术，包括我们最近 在计算机上描述了密码子优化、合成启动子工程和祖先序列重建 技术，以提高目前的理解和更好地评估基因治疗的整体潜力， 适用于所有血友病A患者人群，包括患有fVIII抑制剂的患者。

Project 3: Immunopharmacology of FVIII Bioengineering and Gene Therapy Summary The development of inhibitory antibodies directed against human factor VIII remains the most significant clinical complication associated with the treatment of hemophilia A and is a critical barrier to gene therapy approaches. Therefore, improved understanding of the immunology and pharmacology related to promising liver-directed adeno-associated viral (AAV) vector and hematopoietic stem cell (HSC)-directed lentiviral vector (LV) gene therapy strategies is needed. The studies proposed in Project 3 of this U54 application are designed to advance our knowledge of fVIII immunobiology in the context of these gene therapies. The proposed studies are designed to i) analyze the molecular composition and immune reactivity of bioengineered recombinant infusion products as well as liver-directed AAV and HSC-directed LV gene therapy derived fVIII, ii) identify critical design parameters associated with immunogenicity and inhibitor eradiation potential of liver-directed AAV-fVIII gene therapy and iii) evaluate the immunogenicity and inhibitor eradicating potential of non-genotoxic HSC-directed LV gene therapy protocols. These studies will take advantage of available fVIII, glycomics, structural biology, immunology and gene therapy expertise as well as diverse state-of-the-art technologies, including our recently described in silico codon-optimization, synthetic promoter engineering and ancestral sequence reconstruction technology, to improve the current understanding and better assess the overall potential for gene therapy to address all hemophilia A patient populations including those with fVIII inhibitors.