Molecular and Cellular Characterization of AAV1 Vector

腺相关病毒（AAV）作为基因治疗载体已被广泛研究。 使用重组AAV（rAAV）载体的临床前研究和临床试验的结果表明，该载体用于肌肉基因递送是安全有效的。 目前，最广泛使用的rAAV载体基于AAV血清型2，因为它是最广泛表征的血清型。 最近，AAV血清型1的表征表明，基于AAV 1的载体对于递送到肌肉中比AAV 2有效约10至20倍。 这种显著差异表明，AAV 1载体不仅是AAV 2的替代载体，而且具有优于AAV 2载体的独特优势。 科学上，AAV 1为研究AAV的生物学和媒介学提供了一个很好的工具。 作为我以前工作的延续，提出以下研究以血友病B作为疾病模型来表征AAV 1作为基因治疗载体。 1）。目的：鉴定AAV 1对肌肉的组织嗜性决定因子。 在该特定目的中待测试的假设是AAV 1和AAV 2之间的一些不同氨基酸簇解释了对肌肉的亲和力的差异。 在转导肌肉中的这种显著差异为组织嗜性决定簇提供了可靠的测定。 2）。目的研究AAV 1/2杂合辅助病毒的生物学特性，并鉴定AAV 1病毒的细胞受体。3）。探讨AAV 1及其杂交衍生载体在血友病小鼠中纠正表型的有效性，并将这些载体给予具有抗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.