Molecular and Cellular Characterization of AAV1 Vector

腺相关病毒(AAV)作为一种基因治疗载体已被广泛研究。应用重组AAV(RAAV)载体进行的临床前研究和临床试验结果表明，该载体对于肌肉基因的传递是安全有效的。目前，应用最广泛的rAAV载体是基于AAV血清型2，因为它是特征最广泛的血清型。最近，对AAV1型病毒的鉴定表明，AAV1载体比AAV2载体在肌肉中的输送效率大约高出10到20倍。这些显著的差异表明，AAV1载体不仅是AAV2载体的替代载体，而且比AAV2载体具有独特的优势。从科学上讲，AAV1为研究AAV的生物学和传播学提供了一个很好的工具。作为我先前工作的继续，以下研究旨在以血友病B作为疾病模型来描述AAV1作为基因治疗载体的特征。1)。鉴定AAV1对肌肉的组织趋向性决定因素。在这一特定目的中要检验的假设是，AAV1和AAV2之间的一些不同的氨基酸簇解释了对肌肉亲和力的差异。转导肌肉的这种显著差异为组织趋向性决定因素提供了可靠的分析方法。2)。研究AAV1/2杂合辅助子的生物学特性，鉴定AAV1病毒的细胞受体。3)。目的探讨AAV1及其杂交衍生载体对血友病小鼠表型的纠正作用，并将其应用于中和抗体阳性的动物。

Adeno-associated virus (AAV) has been studied extensively as a gene therapy vector. Results from both preclinical studies and clinical trials using recombinant AAV (rAAV) vector suggest that this vector would be safe and effective for muscle gene delivery. Currently, the most widely used rAAV vector is based on AAV serotype 2 because it is the most extensively characterized serotype. Recently, characterization of AAV serotype 1 demonstrated that AAV1 based vectors are approximately 10 to 20-fold more effective than AAV2 for delivery into muscle. Such dramatic differences suggest that AAV1 vector is not merely an alternative vector for AAV2 but has distinctive advantages over the AAV2 vector. Scientifically, AAV1 offers an excellent tool to study the biology and vectorology of AAV. As a continuation of my previous work, the following studies are proposed to characterize AAV1 as a gene therapy vector using hemophilia B as a disease model. 1). To identify the tissue tropism determinants of AAV1 for muscle. The hypothesis to be tested in this specific aim is that some dissimilar amino acid clusters between AAV1 and AAV2 account for the differences in affinity for muscle. Such dramatic differences in transducing muscle provide a reliable assay for the tissue tropism determinants. 2). To characterize the biological properties of AAV1/2 hybrid helpers, and to identify the cellular receptors for AAV1 virus. 3). To explore the effectiveness of AAV 1 and its hybrid derivative vectors in correcting the phenotype in hemophilic mice, and to administer these vectors to animals with neutralizing antibodies against AAV.