title: Novel cytotoxic vectors based on adeno-associated virus.
abstract: Vectors based on adeno-associated virus (AAV) are promising tools for gene therapy. The production of strongly toxic vectors, for example for cancer-directed gene transfer, is often unfeasible due to uncontrolled expression of toxic genes in vector-producing cells. Using an approach based on transcriptional repression, we have created novel AAV vectors carrying the genes coding for diphtheria toxin A (DTA) and the pro-apoptotic PUMA protein. The DTA vector had a significant toxic effect on a panel of tumor cell lines, and abrogation of protein synthesis could be shown. The PUMA vector had a toxic effect on HeLa and RPMI 8226 cells, and sensitized transduced cells to doxorubicin. To permit targeted gene transfer, we incorporated the DTA gene into a genetically modified AAV-2 capsid previously developed by our group that mediates enhanced transduction of murine breast cancer cells in vitro. This vector had a stronger cytotoxic effect on breast cancer cells than DTA vectors with wildtype AAV capsid or vectors with a random capsid modification. The vector production and application system presented here allows for easy exchange of promotors, transgenes and capsid specificity for certain target cells. It will therefore be of great possible value in a broad range of applications in cytotoxic gene therapy and significantly broadens the spectrum of available tools for AAV-based gene therapy.
authors: Hubertus Wald Cancer Center, Department of Oncology and Hematology, University Medical Center Hamburg-Eppendorf, Martinistr. 52, 20246 Hamburg, Germany. johannes.kohlschuetter@online.de
authors: Johannes
authors: J
authors: Kohlschütter
authors: Stefan
authors: S
authors: Michelfelder
authors: Martin
authors: M
authors: Trepel
mesh_terms: D000818:Animals; D051017:Apoptosis Regulatory Proteins / Q000235:genetics*; D045744:Cell Line, Tumor; D002470:Cell Survival; D000229:Dependovirus / Q000235:genetics*; D004167:Diphtheria Toxin / Q000235:genetics*; D005260:Female; D005822:Genetic Vectors; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D010957:Plasmids; D014407:Tumor Cells, Cultured
chemical_list: D051017:Apoptosis Regulatory Proteins; D004167:Diphtheria Toxin