Molecular Engineering of AAV for Stealth and Targeting

描述（由申请人提供）：基因疗法在治疗和潜在治愈各种疾病方面具有巨大的潜力。然而，在治疗这些疾病之前，基因传递技术需要在安全性、效率和表达稳定性方面有显著的提高。基于腺相关病毒（AAV）的载体在实验室和临床中都被证明是非常有前途的，但它们仍然存在一些缺点。特别是，它们不能轻易地将基因传递到特定的细胞类型，而且它们的效率受到免疫系统对它们的识别和中和的影响。这两种缺陷是由这种病毒的表面特性造成的，我们将采用化学合成和分子生物学相结合的方法来设计病毒表面，以实现靶向基因传递和增强隐身性。具体来说，在病毒衣壳中遗传插入肽标签将允许化学合成的靶向肽和隐形聚合物的模块化和可逆附着。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。确定组合式模块化化学和遗传方法是否能有效靶向AAV基因传递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上解决这一问题。

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.