Engineering AAV capsids for enhanced transduction of skeletal muscle

摘要 我们正在开发一种改进的基于腺相关病毒(AAV)的基因治疗载体 我们称之为“增强型AAV”或简称“eAAV”。EAAV是专门为基因设计的 输送到骨骼肌组织。基于对老鼠的初步研究，我们预计这将是 比使用的重组aAVs(rAAV1和rAAV8)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水平的 治疗性转基因产品，要么直接由于提高了转导效率，要么 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 对转基因产品的反应。 这笔赠款的目的有三个。目标1是在年完成eAAV产品的设计 细胞培养和啮齿动物研究。目标2是在非人类灵长类动物身上测试最终的eAAV产品， 目标3是确定与其他产品相比，它是否构成任何独特的安全风险 RAAV载体。目标2构成了该项目的大部分。在这里，我们将比较eAAV和rAAV 其同时表达α-1抗胰蛋白酶和人类免疫缺陷病毒的能力 (HIV)--来自骨骼肌的抑制因子eCD4。达到治疗水平的AAT将允许 根治遗传性AAT缺乏症，同时获得可靠的高水平eCD4 表达将使艾滋病毒的一次性、可能终身治疗成为可能。

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.