Engineering AAV capsids for enhanced transduction of skeletal muscle

摘要 我们正在开发一种改进的腺相关病毒（AAV）为基础的基因治疗载体 我们称之为“增强型AAV”简称“eAAV”eAAV是专为基因 递送至骨骼肌组织。根据对小鼠的初步研究，我们预计 比用于治疗的重组AAV（rAAV 1和rAAV 8）有效约100倍。 最近的临床试验这种效率的提高应该使我们能够实现更高的水平， 治疗性转基因产物，直接地，由于增加的转导效率，或 间接地，通过允许我们使用更少的载体，从而降低宿主免疫的可能性， 对转基因产物的反应。 这项补助金的目的有三个。目标1是在2010年完成eAAV产品的设计。 细胞培养和啮齿动物研究。目的2是在非人灵长类动物中测试最终的eAAV产物， 目标3是确定与其他产品相比， rAAV载体。目标2是该项目的主要内容。在这里，我们将eAAV与rAAV进行比较， 其表达α-1抗胰蛋白酶（AAT）和人类免疫缺陷病毒的能力 （HIV）-抑制剂eCD 4从骨骼肌。达到AAT的治疗水平将允许 永久治愈遗传性AAT缺陷，同时实现可靠的高水平eCD 4 表达将使艾滋病毒的一次性，潜在的终身治疗成为可能。

ABSTRACT We are developing an improved adeno-associated virus (AAV)-based gene therapy vector that we call “enhanced AAV” or “eAAV” for short. eAAV is specifically designed for gene delivery to skeletal muscle tissue. Based on preliminary studies in mice we expect it will be approximately 100 times more effective than the recombinant AAVs (rAAV1 and rAAV8) used in recent clinical trials. This gain in efficiency should allow us to achieve far higher levels of therapeutic transgene products, either directly, due to increased transduction efficiency, or indirectly, by allowing us to use less vector and thereby reduce the likelihood of host immune responses to the transgene product. The aims of this grant are threefold. Aim 1 is to finalize the design of the eAAV product in cell culture and rodent studies. Aim 2 is to test the final eAAV product in nonhuman primates, and Aim 3 is to determine whether or not it poses any unique safety risks compared to other rAAV vectors. Aim 2 comprises the bulk of the project. Here we will compare eAAV to rAAV for its ability to express both alpha-1 antitrypsin (AAT) and the human immunodeficiency virus (HIV)-inhibitor eCD4 from skeletal muscle. Achieving therapeutic levels of AAT would allow for a permanent cure of genetic AAT-deficiency while achieving reliably high levels of eCD4 expression would enable a one-shot, potentially life-long treatment for HIV.