Novel cytotoxic vectors based on adeno-associated virus
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Novel cytotoxic vectors based on adeno-associated virus. / Kohlschütter, Johannes; Michelfelder, Stefan; Trepel, Martin.
In: Toxins, Vol. 2, No. 12, 12.2010, p. 2754-68.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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TY - JOUR
T1 - Novel cytotoxic vectors based on adeno-associated virus
AU - Kohlschütter, Johannes
AU - Michelfelder, Stefan
AU - Trepel, Martin
PY - 2010/12
Y1 - 2010/12
N2 - 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.
AB - 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.
KW - Animals
KW - Apoptosis Regulatory Proteins
KW - Cell Line, Tumor
KW - Cell Survival
KW - Dependovirus
KW - Diphtheria Toxin
KW - Female
KW - Genetic Vectors
KW - HEK293 Cells
KW - HeLa Cells
KW - Humans
KW - Mice
KW - Plasmids
KW - Tumor Cells, Cultured
KW - Journal Article
KW - Research Support, Non-U.S. Gov't
U2 - 10.3390/toxins2122754
DO - 10.3390/toxins2122754
M3 - SCORING: Journal article
C2 - 22069574
VL - 2
SP - 2754
EP - 2768
JO - Toxins
JF - Toxins
SN - 2072-6651
IS - 12
ER -