Combinatorial selection of efficient and cell-type specific adeno-associated virus vectors (AAV) for human gene therapy

用于人类基因治疗的高效和细胞类型特异性腺相关病毒载体（AAV）的组合选择

• 批准号: 5450151
• 负责人: Professor Dr. Michael Hallek
• 金额: --
• 依托单位: Klinik für Innere Medizin I
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2005
• 资助国家: 德国
• 起止时间: 2004-12-31 至 2007-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/5450151?language=en
• 关键词: Combinatorial; selection; efficient; cell; type

The major goal of this effort is to generate AAV mutants and vectors with a cell or tissue specific tropism. We recently established a novel combinatorial technology, called AAV display, for the production of AAV vectors with redirected tropism. Screening of this library of capsid modified AAV mutants on different cell lines allowed to isolate several viral clones that infect these target cells very efficiently and in a receptor specific manner. The mutants selected so far, however, showed only a limited target selectivity and partially retained their natural tropism. Therefore, the experiments described in this grant proposal aim to further refine this technology. The goal of generating tissue specific AAV vectors will be pursued by optimizing the library selection protocols and by combining this combinatorial approach with an educated mutagenesis of the AAV viral capsid. This educated approach will use the knowledge about the three-dimensional structure of the AAV-2 capsid and introduce distinct modifications in order to - for example - eliminate wild-type receptor binding sites. The AAV retargeting mutants generated by these techniques will be tested extensively both in vitro and in vivo.

这项工作的主要目标是产生具有细胞或组织特异性向性的 AAV 突变体和载体。我们最近建立了一种称为 AAV 显示的新型组合技术，用于生产具有重定向趋向性的 AAV 载体。在不同细胞系上筛选衣壳修饰的 AAV 突变体文库，可以分离出几个病毒克隆，这些病毒克隆可以非常有效地以受体特异性方式感染这些靶细胞。然而，迄今为止选择的突变体仅表现出有限的靶标选择性，并部分保留了其自然趋向性。因此，本拨款提案中描述的实验旨在进一步完善这项技术。将通过优化文库选择方案并将这种组合方法与 AAV 病毒衣壳的受教育诱变相结合来实现生成组织特异性 AAV 载体的目标。这种受过教育的方法将利用有关 AAV-2 衣壳三维结构的知识，并引入不同的修饰，例如消除野生型受体结合位点。通过这些技术产生的 AAV 重定向突变体将在体外和体内进行广泛的测试。