Molecular Engineering of AAV for Stealth and Targeting

描述（由申请人提供）：基因治疗具有治疗和潜在治愈多种疾病的巨大潜力。 然而，基因递送技术需要在安全性、效率和表达稳定性方面的显著改进，才能治疗大多数这些疾病。 基于腺相关病毒（AAV）的载体已被证明在实验室和临床上都是非常有前途的，但它们仍然存在一些缺点。 特别是，它们不能容易地将基因递送靶向特定的细胞类型，并且它们的效率受到免疫系统的识别和中和的影响。 这两个缺点是由于这种病毒的表面特性造成的，我们将实施一种结合化学合成和分子生物学的方法来设计病毒表面，以实现靶向基因递送和增强隐形性。 具体地，将肽标签遗传插入病毒衣壳将允许化学合成的靶向肽和隐形聚合物的模块化和可逆连接。 实际上，这种已经有效的自然产物正在被改造，以更好地满足人类的治疗需求。 具体目标是：1。为了确定组合的模块化化学和遗传方法是否可以有效地靶向AAV基因递送2。为了确定聚乙二醇是否可以从中和抗体中“隐身”AAV，本提案中描述的合成化学、分子生物学和生物工程的新混合物将非常适合在工程和医学的界面处解决这个问题。

DESCRIPTION (provided by applicant): Gene therapy has vast potential for treating and potentially curing a wide variety of disorders. However, gene delivery technologies require significant improvements in safety, efficiency, and expression stability before the majority of these diseases can be treated. Vectors based on adeno-associated virus (AAV) have proven themselves to be highly promising, both in the laboratory and the clinic, but they still suffer from several shortcomings. In particular, they are unable to readily target gene delivery to specific cell types, and their efficiency is compromised by their recognition and neutralization by the immune system. These two liabilities are a result of the surface properties of this virus, and we will implement a combined chemical synthesis and molecular biology approach to engineer the viral surface for targeted gene delivery and enhanced stealth. Specifically, the genetic insertion of a peptide tag into the viral capsid will allow the modular and reversible attachment of chemically synthesized targeting peptides and stealthing polymers. In effect, this already effective product of nature is being engineered to better match human therapeutic needs. The Specific Aims are: 1. To determine whether a combined, modular chemical and genetic approach can effectively target AAV gene delivery 2. To determine whether polyethylene glycol can "stealth" AAV from neutralizing antibodies The novel blend of synthetic chemistry, molecular biology, and bioengineering described in this proposal will be well suited to addressing this problem at the interface of engineering and medicine.