Molecular and Cellular Characterization of AAV1 Vector

腺相关病毒（AAV）作为一种基因治疗载体得到了广泛的研究。利用重组AAV （rAAV）载体进行的临床前研究和临床试验结果表明，该载体可以安全有效地传递肌肉基因。目前，使用最广泛的rAAV载体是基于AAV血清型2，因为它是最广泛表征的血清型。最近，对AAV血清型1的表征表明，基于AAV1的载体比AAV2载体在肌肉中的递送效率高约10至20倍。如此巨大的差异表明AAV1载体不仅仅是AAV2的替代载体，而且比AAV2载体具有明显的优势。在科学上，AAV1为研究AAV的生物学和媒介学提供了一个很好的工具。作为我之前工作的延续，我提出以下研究，以血友病B作为疾病模型来表征AAV1作为基因治疗载体。1）. 确定AAV1在肌肉中的组织趋向性决定因素。在这个特定的目标中需要验证的假设是，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。这种戏剧性的差异在转导肌肉提供了一个可靠的测定组织倾向的决定因素。2）. 目的：研究AAV1/2杂化辅助体的生物学特性，鉴定AAV1病毒的细胞受体。3）. 目的探讨AAV - 1及其杂交衍生载体对血友病小鼠表型的纠正作用，并将这些载体应用于具有AAV中和抗体的动物。

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.