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

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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.