Engineering immuno-evading adeno-associated virus vectors for gene therapy

 描述（由申请人提供）：使用腺相关病毒（AAV）的基因治疗已成为多种疾病（包括那些困扰心血管系统的疾病）的有希望的治疗选择。不幸的是，针对AAV衣壳的体液免疫应答阻止了根据需要重新施用病毒载体的能力。除了抗AAV capsi抗体应答之外，组织靶向是有效心脏基因治疗的另一个主要障碍。在此之前，我们设计了一种基于AAV的蛋白酶激活病毒（PAV），该病毒在病变组织中被过表达的基质金属蛋白酶（MMPs）刺激。尽管PAV可以使更多的靶向递送到疾病部位，但它们也将遭受针对衣壳的抗体应答，就像未工程化的载体一样。为了克服这个问题，我假设，基因插入一个“自我肽”到PAV衣壳（PAV-SP）将最大限度地减少吞噬细胞的摄取，最终减少中和抗体的产生。本研究的主要目的是：1）对PAV-SP进行基因克隆和结构鉴定; 2）利用体外模型对PAV-SP进行功能鉴定。

 DESCRIPTION (provided by applicant): Gene therapy with adeno-associated virus (AAV) has emerged as a promising treatment option for a variety of diseases, including those afflicting the cardiovascular system. Unfortunately, humoral immune responses against the AAV capsid prevent the ability to re-administer the viral vector as needed. In addition to the anti- AAV capsi antibody responses, tissue targeting is another major hurdle to effective cardiac gene therapy. Previously, we designed a Protease-Activatable Virus (PAV) based on AAV that is stimulated by overexpressed matrix metalloproteinases (MMPs) in diseased tissues. Although the PAVs may enable more targeted delivery to sites of disease, they will also suffer from antibody responses against the capsid just like the unengineered vectors. To overcome this problem, I hypothesize that genetically inserting a `self-peptide' into the PAV capsid (PAV-SP) will minimize phagocytic uptake, ultimately decreasing neutralizing antibody production. The aims of this fellowship project are the following: 1) genetically clone and structurally characterize PAV-SP and 2) functionally characterize PAV-SP with in vitro models.