Optimized Adeno-Associated Viral-Mediated Human Factor VIII Gene Therapy in Cynomolgus Macaques
Optimized Adeno-Associated Viral-Mediated Human Factor VIII Gene Therapy in Cynomolgus Macaques
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DOI：
10.1089/hum.2018.080
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发表时间：
2018-07-23
影响因子：
4.2
通讯作者：
Wilson, James M.
中科院分区：
文献类型：
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作者：
Greig, Jenny A.;Nordin, Jayme M. L.;Wilson, James M.
血友病A是一种常见的遗传性出血性疾病，其特征在于缺乏人凝血因子VIII（hFVIII）。先前使用腺相关病毒（AAV）载体的研究鉴定了两种肝脏特异性启动子和增强子组合（E03.TTR和E12.A1AT），其在该疾病的小鼠模型中驱动密码子优化的B结构域缺失的hFVIII转基因的高水平表达。本研究使用两种不同的AAV衣壳（AAVrhlO和AAVhu 37）进一步评估了食蟹猴中的这些增强子/启动子组合。将四种载体组合中的每一种以1.2x10（13）基因组拷贝/kg的剂量静脉内施用到每组五只猕猴中。与施用AAVrh 10载体的动物相比，通过AAVhu 37衣壳递送hFVIII转基因导致hFVIII表达的显著增加。在施用包装在AAVhu 37衣壳内的E03.TTR后两周，平均hFVIII表达为正常的20.2 +/-5.0%，其中一只动物表现出正常hFVIII水平的37.1%的峰值表达。大多数动物在载体递送后第8-10周产生抗hFVIII抗体应答。然而，施用AAVhu37.E03.TTR的五只猕猴中的两只在载体施用后30周内没有可检测的抗体应答。总的来说，该研究支持使用AAVhu 37衣壳继续开发基于AAV的血友病A基因疗法。
Hemophilia A is a common hereditary bleeding disorder that is characterized by a deficiency of human blood coagulation factor VIII (hFVIII). Previous studies with adeno-associated viral (AAV) vectors identified two liver-specific promoter and enhancer combinations (E03.TTR and E12.A1AT) that drove high level expression of a codon-optimized, B-domain-deleted hFVIII transgene in a mouse model of the disease. This study further evaluated these enhancer/promoter combinations in cynomolgus macaques using two different AAV capsids (AAVrh10 and AAVhu37). Each of the four vector combinations was administered intravenously at a dose of 1.2x10(13) genome copy/kg into five macaques per group. Delivery of the hFVIII transgene via the AAVhu37 capsid resulted in a substantial increase in hFVIII expression compared to animals administered with AAVrh10 vectors. Two weeks after administration of E03.TTR packaged within the AAVhu37 capsid, average hFVIII expression was 20.2 +/- 5.0% of normal, with one animal exhibiting peak expression of 37.1% of normal hFVIII levels. The majority of animals generated an anti-hFVIII antibody response by week 8-10 post vector delivery. However, two of the five macaques administered with AAVhu37.E03.TTR were free of a detectable antibody response for 30 weeks post vector administration. Overall, the study supports the continued development of AAV-based gene therapeutics for hemophilia A using the AAVhu37 capsid.